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Rust pathogens cause damage to cereal crops around the world, leading to reduced yield and profit. Current methods of rust control include fungicides, resistant cultivars, and preventative agronomic practices. Some hyperparasites are antagonists of plant pathogens and may provide a potential method of biocontrol against increasingly virulent strains of rust. Very little is known about the mechanism of inhibition of rust growth by hyperparasites, however, isolation of new strains and subsequent characterisation may reveal new treatment strategies for the control of rust. Here we report the isolation of six new fungal hyperparasites and their effects on the development of three Puccinia rust pathogens were examined in vitro, to determine the potential of each as biocontrol agents of rust. Cut-leaf sections of rust-susceptible wheat, barley and oat cultivars were treated with fungal hyperparasite conidia prior to infection with the rust species; Puccinia triticina, P. hordei, and P. coronata f. sp. avenae, respectively. Inhibition of rust spore germination tests were also performed on water agar plates co-inoculated with the isolates. In leaf sections, rust pustule number was significantly (P

Infiltrative cardiomyopathies are a group of disorders characterized by the abnormal deposition of proteins and/or inflammatory cells leading to cardiac dysfunction and electrical disorders. This chapter will illustrate the role of PET/CT in cardiac sarcoidosis and amyloidosis, and the complementary role of PET/MR and 99mTc PYP SPECT.

In 2010, the parasitic fungus Austropuccinia psidii (myrtle rust) was detected in Australia. Austropuccinia psidii infects immature growth of myrtaceous species. Many of Australia’s myrtaceous species occur within fire-prone vegetation communities and have the capacity to resprout after fire. Therefore, it is likely that new post-fire growth may be vulnerable to A. psidii infection, causing subsequent flow-on effects to species’ persistence and community dynamics. The aim of this study was to test the impacts of A. psidii on native Australian Myrtaceae species after fire. We grew eight native susceptible species in a glasshouse experiment before burning them and inoculating the resprouting new growth of half the plants with A. psidii. We assessed the effect of A. psidii on the architecture, growth and biomass allocation of our study species. Although general patterns were observed across species, results were found to be species-specific. Austropuccinia psidii significantly reduced the height of two of the eight species (Callistemon citrinus and Eucalyptus moluccana), but none of the species had increased branching. As expected, specific leaf area was lower (9%) in inoculated plants – although only significant for C. citrinus and E. dalrympleana – and leaf biomass was greater (15%), but significant for Angophora costata only. Finally, biomass allocation did not significantly differ between infection treatments. We can conclude that the effect of A. psidii infection on fire-damaged plants has significant impacts on plants at the species level, which may have flow-on effects at the community level, especially after repeated infections. Furthermore, these impacts may be exacerbated in the future under climate change, as the predicted increase in frequency and intensity of fires across Australia will result in more frequent new growth availability, providing more opportunities for A. psidii infection.

Hemibiotrophic pathogens cause significant losses within agriculture, threatening the sustainability of food systems globally. These microbes colonise plant tissues in three phases: a biotrophic phase followed by a biotrophic-to-necrotrophic switch phase and ending with necrotrophy. Each of these phases is characterized by both common and discrete host transcriptional responses. Plant hormones play an important role in these phases, with foliar models showing that salicylic acid accumulates during the biotrophic phase and jasmonic acid/ethylene responses occur during the necrotrophic phase. The appropriateness of this model to plant roots has been challenged in recent years. The need to understand root responses to hemibiotrophic pathogens of agronomic importance necessitates further research. In this study, using the root hemibiotroph

Long-distance migration and host adaptation by transboundary plant pathogens often brings detrimental effects to important agroecosystems. Efficient surveillance as a basis for responding to the dynamics of such pathogens is often hampered by a lack of information on incursion origin, evolutionary pathways and the genetic basis of rapidly evolving virulence across larger timescales. Here, we studied these genetic features by using historical isolates of the obligate biotrophic pathogen Puccinia striiformis f. sp. tritici (Pst), which causes one of the most widespread and devastating diseases, stripe (yellow) rust, of wheat. Through a combination of genotypic, phenotypic and genomic analyses, we assigned eight Pst isolates representing putative exotic Pst incursions into Australia to four previously defined genetic groups, PstS0, PstS1, PstS10 and PstS13. We showed that isolates of an additional incursion of P. striiformis, known locally as P. striiformis f. sp. pseudo-hordei, had a new and unique multilocus SSR genotype (MLG). We provide results of overall genomic variation of representative Pst isolates from each genetic group by comparative genomic analyses. We showed that isolates within the PstS1 and PstS13 genetic groups are most distinct at the whole-genome variant level from isolates belonging to genetic group PstS0, whereas the isolate from the PstS10 genetic group is intermediate. We further explored variable gene content, including putative effectors, representing both shared but also unique genetic changes that have occurred following introduction, some of which may additionally account for local adaptation of these isolates to triticale. Our genotypic and genomic data revealed new genetic insights into the evolution of diverse phenotypes of rust pathogens following incursion into a geographically isolated continental region.

Austropuccinia psidii (myrtle rust) is an invasive fungus native to South America that infects the young growing tissues of species in the Myrtaceae family, one of the dominant plant families in Australia. To date, 360 native species from 49 genera have been found to be susceptible in Australia, but the vast majority remain untested (81%). The aim of this study was to test a range of plant species whose susceptibility status remains unknown, including endangered species, species with a large distribution overlap with A. psidii and species from a genus that has not been previously tested. Different sub-species and provenances were also tested to assess for intra-specific differences. Of the 24 tested species/sub-species, 18 (including 12 endangered) were found susceptible to A. psidii to varying degrees (including the first species record within Triplarina being susceptible), while one presented a hypersensitive reaction and six were resistant. The most susceptible species were the critically endangered Melaleuca megalongensis, and the endangered Eucalyptus copulans, E. parvula, E. scoparia and Melaleuca irbyana. No significant differences in intraspecific susceptibility were found between sub-species or provenances. We suggest that the susceptible species we have identified will be vulnerable to infection in their native ranges in the future, if they have not already become infected. For highly susceptible species, A. psidii should be considered as a major additional threat and appropriate control measures incorporated into existing threatened species plans. Monitoring of susceptible species’ populations in the wild and seed collection for seed banking are vital steps for ensuring their conservation.

Hypobaria and hypoxia exert independent effects on oxidative stress during exercise, while combined effectson the post-exercise recovery period remain unclear.Accordingly, this study examined the recovery period during lab-simulated hypoxic and hypobaric conditions following exercise-induced oxidative stress. Participants (n=13) performed 60-minutes of cycling (70% watts max) in a normobaric normoxic environment followed by a four-hour recovery under three conditions; 1000m normobaric normoxia (NN, 675mmHg), 4400m normobaric hypoxia (NH, 675mmHg), or 4400m hypobaric hypoxia (HH, 440mmHg). Blood samples collected at Pre, Post, 2-Hours (2-HR), and 4-Hours (4-HR) post-exercise were analyzed fora potential increase in biochemical modifications of proteins(protein carbonyls, PC; 3-nitrotyrosines, 3NT) lipids (lipid hydroperoxides, LOOH; 8-isoprostanes, 8-ISO), and antioxidant capacity (FRAP, TEAC). Gene transcripts (EPAS, HMOX1, SOD2, NFE2L2) were quantified by qRT-PCR from muscle biopsies taken Pre and Post exercise. Hypoxia and hypobaria had no effect throughout recovery. Post-exercise TEAC (p=0.041), FRAP (p=0.013), and 8-ISO (p=0.044) increased, while PC (p=0.002) and 3-NT (p=0.032) were decreased. LOOH was lower in Post (p=0.018) NH trial samples. Exercise-dependent increases occurred in NFE2L2 (p=0.003), HMXO1 (p0.001), SOD2 (p=0.046), and EPAS (p=0.038). Exercise recovery under conditions of NH and HH did not impact blood oxidative stress or redox-sensitive gene transcripts.

Myrtle rust, caused by the pathogen Austropuccinia psidii, affects species of the Myrtaceae, many of which are endemic to Australia and New Zealand. Originating from South America, A. psidii is now present in both countries, necessitating effective chemical control for disease management. Using an artificial inoculation protocol, the efficacy of eight fungicides (tebuconazole/trifloxystrobin, cyproconazole/azoxystrobin, fosetyl aluminum, triforine, triadimenol, oxycarboxin, copper, and tebuconazole) applied as curative or protectant treatments was tested on two native New Zealand species (Lophomyrtus × ralphii and Metrosideros excelsa). The impacts of rate (×2), frequency (single or double), and timing (pre- or postinfection) of fungicide application were investigated. Overall, the most effective fungicides tested across both species were those that included a demethylation inhibitor and strobilurin mix, notably tebuconazole/trifloxystrobin (Scorpio) and cyproconazole/azoxystrobin (Amistar Xtra). These fungicides significantly reduced infection of host plants relative to the water control. Timing of application significantly affected bioefficacy, with applications made 7 days before inoculation or 7 days after inoculation being generally the most effective. The rate of fungicide application was not significant for both host species, with few interaction terms showing overall significance. Key findings from this study will set the foundation for further fungicide bioefficacy research conducted to evaluate formulations and adjuvant mixtures, determine suitable application methods for enhanced retention and coverage, and derive optimum application time for effective protection of native and exotic Myrtaceae species in New Zealand.

Verticillium dahliae, the causal agent of Verticillium wilt, is a soil-borne ascomycete that infects numerous agriculturally important crops globally, including cotton. As a billion-dollar industry, cotton is economically important to Australia and the management of disease such as Verticillium wilt is key for the success of the industry. Internationally, defoliating V. dahliae isolates belonging to Vegetative Compatibility Group (VCG) 1A cause severe damage to cotton, while non-defoliating VCG2A isolates result in significantly less disease. However, in Australia, VCG2A is causing more severe damage to crops in the field than the defoliating VCG1A. This study aimed to replicate field observations in controlled greenhouse conditions. We examined and compared disease symptoms on a range of Australian commercial cotton varieties when inoculated with different V. dahliae VCGs. Seedlings were root dipped in conidial suspensions and assessed over seven weeks. The final disease score, disease over time and root length were analysed. Plant mortality resulted from both V. dahliae VCG1A and VCG2A isolates across all cotton varieties used, confirming that there are virulent VCG2A isolates present in Australia. To our knowledge, although virulent on other plant hosts, V. dahliae VCG2A has not previously been reported to be highly virulent in cotton. We infer that virulence cannot be defined solely by VCG in Australian V. dahliae isolates causing disease in cotton.

Rust diseases are serious threats to New Zealand cereal crops. Beside the use of fungicides, resistant varieties are an important option for managing these diseases. Changes in rust pathotypes commonly occur due to mutations in existing populations or exotic incursions. Information on these changes is the basis of gene-based disease management. Rust-infected leaves were collected from cereal crops from 2012 to 2015. The pathotypes of these and some historic samples were determined in glasshouse studies, using specific differential host sets. Eight pathotypes of Puccinia triticina (Pt, causal agent of wheat leaf rust), five of P. striiformis f. sp. tritici (Pst, causal agent of wheat stripe rust) and two of P. hordei (Ph, causal agent of barley leaf rust) were identified. The Pst ‘WA’ pathotype was most frequently found. Wheat varieties ‘Empress’ and ‘Torch’, previously resistant to Pt, were found to be susceptible to leaf rust for the first time. The ‘WA’ pathotype of Pst is likely to have arrived in New Zealand from Australia, and is now widespread. The two Pt pathotypes could have overcome resistance gene Lr24 in ‘Empress’ and ‘Torch’.

Background The prevalence of azole resistance in Aspergillus fumigatus is uncertain in Australia. Azole exposure may select for resistance. We investigated the frequency of azole resistance in a large number of clinical and environmental isolates. Methods A. fumigatus isolates [148 human, 21 animal and 185 environmental strains from air (n = 6) and azole-exposed (n = 64) or azole-naive (n = 115) environments] were screened for azole resistance using the VIPcheck™ system. MICs were determined using the Sensititre™ YeastOne YO10 assay. Sequencing of the Aspergillus cyp51A gene and promoter region was performed for azole-resistant isolates, and cyp51A homology protein modelling undertaken. Results Non-WT MICs/MICs at the epidemiological cut-off value of one or more azoles were observed for 3/148 (2%) human isolates but not amongst animal, or environmental, isolates. All three isolates grew on at least one azole-supplemented well based on VIPcheck™ screening. For isolates 9 and 32, the itraconazole and posaconazole MICs were 1 mg/L (voriconazole MICs 0.12 mg/L); isolate 129 had itraconazole, posaconazole and voriconazole MICs of >16, 1 and 8 mg/L, respectively. Soil isolates from azole-exposed and azole-naive environments had similar geometric mean MICs of itraconazole, posaconazole and voriconazole (P > 0.05). A G54R mutation was identified in the isolates exhibiting itraconazole and posaconazole resistance, and the TR34/L98H mutation in the pan-azole-resistant isolate. cyp51A modelling predicted that the G54R mutation would prevent binding of itraconazole and posaconazole to the haem complex. Conclusions Azole resistance is uncommon in Australian clinical and environmental A. fumigatus isolates; further surveillance is indicated.

Background. The presence of patent foramen ovale (PFO) has been linked to many illness, including cryptogenic stroke, transient ischemic attack, migraine, platypnea-orthodeoxia syndrome and decompression sickness in scuba divers. Transesophageal echocardiography is the gold standard technique for the visualization of atrial septal anatomy, but it is a secondary level exam, not always available, with additional associated costs and not completely free from procedural risks. Standard transthoracic echocardiography (TTE) has a too low sensitivity for PFO screening. Purpose. The aim of the study was to assess the role of TTE associated with agitated saline contrast injection (contrast-TTE) as a gatekeeper for the identification of PFO in a large cohort of patients undergoing selection for percutaneous closure. Methods. A total of 200 patients undergoing a diagnostic work-up for the identification of PFO was imaged by contrast-TTE at rest and after provocative maneuvers (PM: Valsalva in all cases). Contrast TTE was graded from 0 to 4 on the bases of bubbles counting (0: no bubbles; 1: 30 bubbles; 4: complete LV opacification). PFO closure was performed after a consensual clinical decision by the cardiologist and the neurologist taking into account comprehensive imaging, clinical evaluation and thrombophilia screening. PFO closure was always monitored by intracardiac echocardiography. Results. At baseline contrast TTE was positive (≥2) in 34 patients (17%) while contrast TTE with PM was positive in 94 cases (47%). 27 out of 200 patients (14%) had an interatrial septal aneurysms. PFO closure was performed in 34 cases (17%). All of these had severe right-to-left shunting (≥3) at contrast TTE and 9 cases had also an interatrial septal aneurysms. The procedure was aborted in only 1 patient due to a complex defect anatomy. Conclusion. Contrast TTE with PM may be not only considered an accurate tool for the detection of PFO but may be also inserted in the diagnostic work- up as a primary gatekeeper for percutaneous closure. Severe shunting at contrast TTE influences final decision making in a large cohort of cases undergoing screening for PFO closure.

This study aimed to determine the physiological and thermoregulatory responses of individuals exercising in the heat (US military red flag conditions, wet-bulb globe temperature 31.5-32.2ºC) while consuming varied volumes of ambient temperature water and ice slurry.Participants (N = 12) walked on a treadmill for 3 hours at approximately 40% peak aerobic capacity in a hot environment while consuming ambient temperature (35.5°C) water (W), ice slurry (0°C, two-thirds shaved ice and one-third water) at a ratio of 2 g·kg(-1) body mass every 10 minutes (FS), and reduced volume ice slurry as described at a rate of 1 g·kg(-1) body mass every 10 minutes (HS). Trials were completed at least 14 days apart, in a randomized, repeated measures design.Percent body weight loss was higher during the HS trial (1.8 ± 0.01%) compared with FS (0.5 ± 0.01%; P.001) and W (0.6 ± 0.01%; P.001). Mean rectal temperature at 3 hours was lower during FS (37.8 ± 0.7°C) compared with HS (38.1 ± 0.8°C) and W (38.2 ± 0.8°C) (P = .04 vs HS, and P = .005 vs W, main effect for trial). No differences were found in rectal temperature between HS and W. Heart rate was lower at the end of the third hour during FS (141 ± 10 beats/min) compared with HS (157 ± 19 beats/min) and W (154 ± 18 beats/min) (P = .001 and P = .007, respectively, time × trial interaction). There were no differences in heart rate between HS and W.The temperature of consumed fluids may be as important as the volume for the management of thermoregulation and other physiological responses for extended work in hot environments.

A long-standing biological question is how evolution has shaped the genomic architecture of dikaryotic fungi. To answer this, high-quality genomic resources that enable haplotype comparisons are essential. Short-read genome assemblies for dikaryotic fungi are highly fragmented and lack haplotype-specific information due to the high heterozygosity and repeat content of these genomes. Here, we present a diploid-aware assembly of the wheat stripe rust fungus Puccinia striiformis f. sp. tritici based on long reads using the FALCON-Unzip assembler. Transcriptome sequencing data sets were used to infer high-quality gene models and identify virulence genes involved in plant infection referred to as effectors. This represents the most complete Puccinia striiformis f. sp. tritici genome assembly to date (83 Mb, 156 contigs, N50 of 1.5 Mb) and provides phased haplotype information for over 92% of the genome. Comparisons of the phase blocks revealed high interhaplotype diversity of over 6%. More than 25% of all genes lack a clear allelic counterpart. When we investigated genome features that potentially promote the rapid evolution of virulence, we found that candidate effector genes are spatially associated with conserved genes commonly found in basidiomycetes. Yet, candidate effectors that lack an allelic counterpart are more distant from conserved genes than allelic candidate effectors and are less likely to be evolutionarily conserved within the P. striiformis species complex and Pucciniales. In summary, this haplotype-phased assembly enabled us to discover novel genome features of a dikaryotic plant-pathogenic fungus previously hidden in collapsed and fragmented genome assemblies., IMPORTANCE Current representations of eukaryotic microbial genomes are haploid, hiding the genomic diversity intrinsic to diploid and polyploid life forms. This hidden diversity contributes to the organism’s evolutionary potential and ability to adapt to stress conditions. Yet, it is challenging to provide haplotype-specific information at a whole-genome level. Here, we take advantage of long-read DNA sequencing technology and a tailored-assembly algorithm to disentangle the two haploid genomes of a dikaryotic pathogenic wheat rust fungus. The two genomes display high levels of nucleotide and structural variations, which lead to allelic variation and the presence of genes lacking allelic counterparts. Nonallelic candidate effector genes, which likely encode important pathogenicity factors, display distinct genome localization patterns and are less likely to be evolutionary conserved than those which are present as allelic pairs. This genomic diversity may promote rapid host adaptation and/or be related to the age of the sequenced isolate since last meiosis.

The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand1. This slow improvement rate is attributed partly to the long generation times of crop plants. Here, we present a method called ‘speed breeding’, which greatly shortens generation time and accelerates breeding and research programmes. Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum) and pea (Pisum sativum), and 4 generations for canola (Brassica napus), instead of 2–3 under normal glasshouse conditions. We demonstrate that speed breeding in fully enclosed, controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies and transformation. The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent (SSD) and potential for adaptation to larger-scale crop improvement programs. Cost saving through light-emitting diode (LED) supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing and genomic selection, accelerating the rate of crop improvement.

Lecanicillium psalliotae is an entomopathogenic, mycoparasitical, and nematophagous fungus known to produce antibiotic and antifungal compounds. Here, we report the first 36-Mb draft genome sequence of L. psalliotae strain HWLR35. The draft genome contains 197 scaffolds and is predicted to have 11,009 protein-coding genes.

A variety of dietary choices are marketed to enhance glycogen recovery after physical activity. Past research informs recommendations regarding the timing, dose, and nutrient compositions to facilitate glycogen recovery. This study examined the effects of isoenergetic sport supplements (SS) vs. fast food (FF) on glycogen recovery and exercise performance. Eleven males completed two experimental trials in a randomized, counterbalanced order. Each trial included a 90-min glycogen depletion ride followed by a 4-hr recovery period. Absolute amounts of macronutrients (1.54 ± 0.27 g·kg-1 carbohydrate, 0.24 ± 0.04 g·kg fat-1, and 0.18 ± 0.03g·kg protein-1) as either SS or FF were provided at 0 and 2 hr. Muscle biopsies were collected from the vastus lateralis at 0 and 4 hr post exercise. Blood samples were analyzed at 0, 30, 60, 120, 150, 180, and 240 min post exercise for insulin and glucose, with blood lipids analyzed at 0 and 240 min. A 20k time-trial (TT) was completed following the final muscle biopsy. There were no differences in the blood glucose and insulin responses. Similarly, rates of glycogen recovery were not different across the diets (6.9 ± 1.7 and 7.9 ± 2.4 mmol·kg wet weight- 1·hr-1 for SS and FF, respectively). There was also no difference across the diets for TT performance (34.1 ± 1.8 and 34.3 ± 1.7 min for SS and FF, respectively. These data indicate that short-term food options to initiate glycogen resynthesis can include dietary options not typically marketed as sports nutrition products such as fast food menu items.

Altitude exposure and exercise elicit oxidative stress in blood; however, exercise recovery at 5000 m attenuates oxidative stress. The purpose was to determine the altitude threshold at which blood oxidative stress is blunted during exercise recovery. Twelve males 18-28 years performed four-cycle ergometry bouts (60 min, 70% VO2max, at 975 m). In a randomised counterbalanced crossover design, participants recovered 6 h at 0, 1667, 3333 and 5000 m in a normobaric hypoxia chamber (recovery altitudes were simulated by using a computerised system in an environmental chamber by lowering the partial pressure of oxygen to match that of the respective altitude). Oxygen saturation was monitored throughout exercise recovery. Blood samples obtained pre-, post-, 1 h post- and 5 h post-exercise were assayed for ferric-reducing antioxidant plasma, Trolox equivalent antioxidant capacity, uric acid, lipid hydroperoxides and protein carbonyls. Muscle biopsies obtained pre and 6 h were analysed by real-time polymerase chain reaction to quantify expression of hemeoxgenase 1, superoxide dismutase 2 and nuclear factor (euthyroid-derived 2)-like factor. Pulse oximetry data were similar during exercise, but decreased for the three highest recovery elevations (0 m = 0%, 1667 m = -3%; 3333 m = -7%; 5000 m = -17%). A time-dependent oxidative stress occurred following exercise for all variables, but the two highest recovery altitudes partially attenuated the lipid hydroperoxide response (0 m = +135%, 1667 m = +251%, 3333 m = +99%; 5000 m = +108%). Data may indicate an altitude threshold between 1667 and 3333 m, above which the oxidative stress response is blunted during exercise recovery.

The purpose of this study was to investigate the effects of pneumatic compression pants on postexercise glycogen resynthesis. Active male subjects (n = 10) completed 2 trials consisting of a 90-minute glycogen depleting ride, followed by 4 hours of recovery with either a pneumatic compression device (PCD) or passive recovery (PR) in a random counterbalanced order. A carbohydrate beverage (1.8 g·kg bodyweight) was provided at 0 and 2 hours after exercise. Muscle biopsies (vastus lateralis) were obtained immediately and 4 hours after exercise for glycogen analyses. Blood samples were collected throughout recovery to measure glucose and insulin. Eight fingerstick blood samples for lactate were collected in the last 20 minutes of the exercise period and during the initial portion of the recovery period. Heart rate was monitored throughout the trial. During the PCD trial, subjects recovered using a commercially available recovery device (NormaTec PCD) operational at 0-60 and 120-180 minutes into recovery period. The same PCD was worn during the PR trial but was not turned on to create pulsatile pressures. There was no difference in muscle glycogen resynthesis during the recovery period (6.9 ± 0.8 and 6.9 ± 0.5 mmol·kg wet wt·h for the PR and PCD trials, respectively). Blood glucose, insulin, and lactate concentrations changed with respect to time but were not different between trials (p > 0.05). The use of PCD did not alter the rate of muscle glycogen resynthesis, blood lactate, or blood glucose and insulin concentrations associated with a postexercise oral glucose load.

It is unclear whether the capabilities of recreationally active modern humans are unique due to present training practices or linked to the selection resulting from migration, escape, scavenging, and hunting and/or endurance running in early Homo. The purpose of this study was to determine upper values for total energy expenditure (TEE) and water turnover during sustained work for periods of 12-24 and 12-48 h, respectively and compare them with other species and proposed activities of early Homo. Stable isotopic water (2H218O) was used during competitions in hot environments to establish energy expenditure rates of approaching 10 times resting metabolism (RM) for 12.7 and 26.8 h, respectively. These events demonstrate pronounced hydration demands, with water output rates ranging from 25–95% of initial total body water for events lasting 12–48 h, respectively. These results provide new evidence for a high, sustained work (0.5–2 days) output and hydration demand in humans compared to other species in hot environments. Although the span for sustained metabolic activity in humans is large, it does not require elite level training status so long as adequate exogenous fuel and water are accessible. Because these values far exceed reported expectations/needs for foraging and persistence hunting in early Homo, it remains unclear if the phenomenon of the metabolic range is a modern human characteristic. While modern recreational endurance participants can demonstrate a TEE approaching 10 times RM, the rationale and need for such a high human metabolic ceiling is unclear when considering the energy demands of early Homo.

Background The ability to physically perform at high altitude may require unique strategies to acclimatize before exposure. The effect of acute hypoxic exposure on the metabolic response of the skeletal muscle may provide insight into the value of short-term preacclimatization strategies. Objective To determine the human skeletal muscle response to a single acute bout of exercise in a hypoxic environment on metabolic gene expression. Methods Eleven recreationally active male participants (24 ± 4 years, 173 ± 20 cm, 82 ± 12 kg, 15.2 ± 7.1% fat, 4.0 ± 0.6 L/min maximal oxygen consumption) completed two 1-hour cycling exercise trials at 60% of peak power followed by 4 hours of recovery in ambient environmental conditions (975 m) and at normobaric hypoxic conditions simulating 3000 m in a randomized counterbalanced order. Muscle biopsies were obtained from the vastus lateralis before exercise and 4 hours after exercise for real-time polymerase chain reaction analysis of select metabolic genes. Results Gene expression of hypoxia-inducible factor 1 alpha, cytochrome c oxidase subunit 4, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, hexokinase, phosphofructokinase, mitochondrial fission 1, and mitofusin-2 increased with exercise ( P P > .05). Optic atrophy 1 did not increase with exercise or differ between environmental conditions ( P > .05). Conclusions The improvements in mitochondrial function reported with intermittent hypoxic training may not be explained by a single acute hypoxic exposure, and thus it appears that a longer period of preacclimatization than a single exposure may be required.

A long-standing biological question is how evolution has shaped the genomic architecture of dikaryotic fungi. To answer this, high quality genomic resources that enable haplotype comparisons are essential. Short-read genome assemblies for dikaryotic fungi are highly fragmented and lack haplotype-specific information due to the high heterozygosity and repeat content of these genomes. Here we present a diploidaware assembly of the wheat stripe rust fungusPuccinia striiformisf. sp.triticibased on long-reads using the FALCON-Unzip assembler. RNA-seq datasets were used to infer high quality gene models and identify virulence genes involved in plant infection referred to as effectors. This represents the most completePuccinia striiformisf. sp.triticigenome assembly to date (83 Mb, 156 contigs, N50 1.5 Mb) and provides phased haplotype information for over 92% of the genome. Comparisons of the phase blocks revealed high inter-haplotype diversity of over 6%. More than 25% of all genes lack a clear allelic counterpart. When investigating genome features that potentially promote the rapid evolution of virulence, we found that candidate effector genes are spatially associated with conserved genes commonly found in basidiomycetes. Yet candidate effectors that lack an allelic counterpart are more distant from conserved genes than allelic candidate effectors, and are less likely to be evolutionarily conserved within theP. striiformisspecies complex andPucciniales. In summary, this haplotype-phased assembly enabled us to discover novel genome features of a dikaryotic plant pathogenic fungus previously hidden in collapsed and fragmented genome assemblies.ImportanceCurrent representations of eukaryotic microbial genomes are haploid, hiding the genomic diversity intrinsic to diploid and polyploid life forms. This hidden diversity contributes to the organism’s evolutionary potential and ability to adapt to stress conditions. Yet it is challenging to provide haplotype-specific information at a whole-genome level. Here, we take advantage of long-read DNA sequencing technology and a tailored-assembly algorithm to disentangle the two haploid genomes of a dikaryotic pathogenic wheat rust fungus. The two genomes display high levels of nucleotide and structural variations, which leads to allelic variation and the presence of genes lacking allelic counterparts. Non-allelic candidate effector genes, which likely encode important pathogenicity factors, display distinct genome localization patterns and are less likely to be evolutionary conserved than those which are present as allelic pairs. This genomic diversity may promote rapid host adaptation and/or be related to the age of the sequenced isolate since last meiosis.

The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand [1]. This slow improvement rate is attributed partly to the long generation times of crop plants. Here we present a method called ‘speed breeding’, which greatly shortens generation time and accelerates breeding and research programs. Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum), and pea (Pisum sativum) and 4 generations for canola (Brassica napus), instead of 2-3 under normal glasshouse conditions. We demonstrate that speed breeding in fully-enclosed controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies, and transformation. The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent and potential for adaptation to larger-scale crop improvement programs. Cost-saving through LED supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing, and genomic selection, accelerating the rate of crop improvement.

The purpose of this study was to determine the impact of the core to skin temperature gradient during incremental running to volitional fatigue across varying environmental conditions. A secondary aim was to determine if a "critical" core temperature would dictate volitional fatigue during running in the heat. 60 participants (n=49 male, n=11 female; 24±5 yrs, 177±11 cm, 75±13 kg) completed the study. Participants were uniformly stratified into a specific exercise temperature group (18 °C, 26 °C, 34 °C, or 42 °C) based on a 3-mile run performance. Participants were equipped with core and chest skin temperature sensors and a heart rate monitor, entered an environmental chamber (18 °C, 26 °C, 34 °C, or 42 °C), and rested in the seated position for 10 min before performing a walk/run to volitional exhaustion. Initial treadmill speed was 3.2 km h(-1) with a 0% grade. Every 3 min, starting with speed, speed and grade increased in an alternating pattern (speed increased by 0.805 km h(-1), grade increased by 0.5%). Time to volitional fatigue was longer for the 18 °C and 26 °C group compared to the 42 °C group, (58.1±9.3 and 62.6±6.5 min vs. 51.3±8.3 min, respectively, p

Hypoxia due to altitude diminishes performance and alters exercise oxidative stress responses. While oxidative stress and exercise are well studied, the independent impact of hypoxia on exercise recovery remains unknown. Accordingly, we investigated hypoxic recovery effects on post-exercise oxidative stress. Physically active males (n = 12) performed normoxic cycle ergometer exercise consisting of ten high:low intensity intervals, 20 min at moderate intensity, and 6 h recovery at 975 m (normoxic) or simulated 5,000 m (hypoxic chamber) in a randomized counter-balanced cross-over design. Oxygen saturation was monitored via finger pulse oximetry. Blood plasma obtained pre- (Pre), post- (Post), 2 h post- (2Hr), 4 h post- (4Hr), and 6 h (6Hr) post-exercise was assayed for Ferric Reducing Ability of Plasma (FRAP), Trolox Equivalent Antioxidant Capacity (TEAC), Lipid Hydroperoxides (LOOH), and Protein Carbonyls (PC). Biopsies from the vastus lateralis obtained Pre and 6Hr were analyzed by real-time PCR quantify expression of Heme oxygenase 1 (HMOX1), Superoxide Dismutase 2 (SOD2), and Nuclear factor (euthyroid-derived2)-like factor (NFE2L2). PCs were not altered between trials, but a time effect (13 % Post-2Hr increase, p = 0.044) indicated exercise-induced blood oxidative stress. Plasma LOOH revealed only a time effect (p = 0.041), including a 120 % Post-4Hr increase. TEAC values were elevated in normoxic recovery versus hypoxic recovery. FRAP values were higher 6Hr (p = 0.045) in normoxic versus hypoxic recovery. Exercise elevated gene expression of NFE2L2 (20 % increase, p = 0.001) and SOD2 (42 % increase, p = 0.003), but hypoxic recovery abolished this response. Data indicate that recovery in a hypoxic environment, independent of exercise, may alter exercise adaptations to oxidative stress and metabolism.

To evaluate the previously developed physiological strain index (PSI) model using heart rate and skin temperature to provide further insight into the detection and estimation of thermal and physiological heat strain indices. A secondary aim was to characterize individuals who excel in their performance in the heat.56 male participants completed 2 walking trials (3.5 miles per hour, 5% grade) in controlled environments of 43.3 °C and 15.5 °C (40% humidity). Core and skin temperature, along with heart rate and PSI, were continually monitored during exercise. Participants completed a physical fitness test.The logistic regression model exhibited 4 false positives and 1 false negative at the 40% decision boundary. The "Not at Risk" group (N = 33) had higher body weight (84 ± 13 vs. 77 ± 10 kg, respectively) compared to the "At Risk" (N = 23) group, p0.05. The "Not at Risk" group had a faster 3-mile run time compared to the "At Risk" group (21:53 ± 3:13 vs. 25:16 ± 2:37, respectively), p0.05. During the Heat Trial, the "At Risk" group had a higher rating of perceived exertion at 60 and 90 minutes compared to the "Not at Risk" group (13.5 ± 2.8 vs. 11.5 ± 1.8 and 14.8 ± 3.2 vs. 12.2 ± 2.0 for "At Risk" vs. "Not at Risk" at 60 and 90 minutes, respectively), p0.05.The previously developed model relating heart rate and skin temperature to PSI is highly accurate at assessing heat risk status. Participants classified as "At Risk" had lower physical performance scores and different body weights compared to the "Not at Risk" group and perceived themselves as working harder during exercise in the heat.

The purpose of this investigation was to determine the impact of post-exercise environmental cold exposure on muscle glycogen, PGC-1α, and downstream transcription factors.Eight males cycled for 1h and recovered in either 7 °C (cold) or 20 °C (room temp) environment for 4h. Muscle biopsies were obtained pre, post, and 4h post exercise for the analysis of muscle glycogen and mRNA. During recovery participants consumed 1.8 g kg⁻¹ of body weight of an oral dextrose solution immediately following the post biopsy and 2h into recovery. Blood samples were obtained post exercise and at 30, 60, 120, 150, 180, and 240 min post exercise for the analysis of serum glucose and insulin AUC.Oxygen uptake was lower during room temp than during cold recovery (0.40 ± 0.05 L x min⁻¹ vs. 0.80 ± 0.12 L x min⁻¹; p0.01). There was no effect of temperature on muscle glycogen recovery or glucose AUC. However, insulin AUC was greater during the room temp trial compared to the cold trial (5139 ± 1412 vs. 4318 ± 1272, respectively; p=0.025). PGC-1α gene expression was higher (p=0.029), but ERRα and NRF2 were lower (p=0.019 and p=0.046, respectively) after recovery in the cold. There were no differences in NRF1 (p=.173) or TFAM (p=0.694).This investigation shows no effect of a cold recovery environment on glycogen re-synthesis but does demonstrate reduced ERRα and NRF2 mRNA despite elevations in PGC-1α mRNA when recovery post-exercise takes place in a cold environment.

This study investigated the effect of cyanobacterial inoculants on salt tolerance in wheat. Unicyanobacterial crusts of Nostoc, Leptolyngbya and Microcoleus were established in sand pots. Salt stress was targeted at 6 and 13 dS m−1, corresponding to the wheat salt tolerance and 50 % yield reduction thresholds, respectively. Germinated wheat seeds were planted and grown for 14 (0 and 6 dS m−1) and 21 (13 dS m−1) days by which time seedlings had five emergent leaves. The effects of cyanobacterial inoculation and salinity on wheat growth were quantified using chlorophyll fluorescence, inductively coupled plasma-optical emission spectrometry and biomass measurements. Chlorophyll fluorescence was negatively affected by soil salinity and no change was observed in inoculated wheat. Effective photochemical efficiency correlated with a large range of plant nutrient concentrations primarily in plant roots. Inoculation negatively affected wheat biomass and nutrient concentrations at all salinities, though the effects were fewer as salinity increased. The most likely explanation of these results is the sorption of nutrients to cyanobacterial extracellular polymeric substances, making them unavailable for plant uptake. These results suggest that cyanobacterial inoculation may not be appropriate for establishing wheat in saline soils but that cyanobacteria could be very useful for stabilising soils.

OBJECTIVE.-: Hydration is an important logistical consideration for persons performing in austere environments because water demands must be balanced with the burden of carrying water. METHODS.-: Seven novice climbers participated in a study to determine the hydration kinetics and core temperatures associated with a successful summit of Mount Rainier. Ingestible radio-equipped thermometer capsules were swallowed to monitor core temperature, and an oral dose of deuterium (0.12 ± 0.02 g·kg(-1) body weight) was administered to determine hydration kinetics. RESULTS.-: Mean core temperature throughout the 5.5-hour climb to Camp Muir (3000 m) was 37.6 ± 0.3°C. Water turnover was 95.0 ± 17.5 mL·kg(-1)·24 h(-1) over the duration of the 43-hour study. There was a trend for reduced body mass from before (75.9 ± 13.0 kg) to after (74.8 ± 12.5 kg) the climb (P = .06), and urine specific gravity increased from before (1.013 ± 0.002) to after (1.022 ± 0.006) the climb (P = .004). CONCLUSIONS.-: Hydration demands of climbing Mount Rainier are highly elevated despite modest fluctuations in core temperature. Participants experienced hypohydration but were able to maintain sufficient hydration to successfully summit Mount Rainier and return home safely. Language: en

A real-time thermoregulatory model using noninvasive measurements as inputs was developed for predicting physiological responses of individuals working long hours. The purpose of the model is to reduce heat-related injuries and illness by predicting the physiological effects of thermal stress on individuals while working. The model was originally validated mainly by using data from controlled laboratory studies. This study expands the validation of the model with field data from 26 test volunteers, including US Marines, Australian soldiers, and US wildland fire fighters (WLFF). These data encompass a range of environmental conditions (air temperature: 19-30° C; relative humidity: 25-63%) and clothing (i.e., battle dress uniform, chemical-biological protective garment, WLFF protective gear), while performing diverse activities (e.g., marksmanship, marching, extinguishing fires, and digging). The predicted core temperatures (Tc), calculated using environmental, anthropometric, clothing, and heart rate measures collected in the field as model inputs, were compared with subjects' Tc collected with ingested telemetry temperature pills. Root mean standard deviation (RMSD) values, used for goodness of fit comparisons, indicated that overall, the model predictions were in close agreement with the measured values (grand mean of RMSD: 0.15-0.38° C). Although the field data showed more individual variability in the physiological data relative to more controlled laboratory studies, this study showed that the performance of the model was adequate.

Does the diversity of cyanobacteria in the cycad rhizosphere relate to the cyanobiont species found in the coralloid roots of these ancient plants? The aim of this study was to identify the diversity of soil cyanobacteria occurring in the immediate vicinity of 22 colonized coralloid roots belonging to members of the cycad genera: Macrozamia, Lepidozamia, Bowenia and Cycas. The majority of coralloid roots were sampled at depths > 10 cm below the soil surface. A total of 32 cyanobacterial isolates were cultured and their 16S rRNA gene partially sequenced. Phylogenetic analysis revealed nine operational taxonomic units of soil cyanobacteria comprising 30 Nostoc spp., a Tolypothrix sp. and a Leptolyngbya sp. Microscopy indicated that all isolates were unialgal and confirmed their genus identity. Rhizospheric diversity was compared to existing data on cyanobionts isolated at the same time from the cycad coralloid root. The same isolate was present in both the cycad coralloid root and rhizosphere at only six sites. Phylogenetic evidence indicates that most rhizosphere isolates were distinct from root cyanobionts. This weak relationship between the soil cyanobacteria and cycad cyanobionts might indicate that changes in the soil community composition are due to environmental factors.

The purpose of this study was to assess the physiological adaptations in physically fit individuals to a period of intensified training. Ten trained males cycled outdoors ~170 km day(-1) on 19 out of 21 days. Expired gas was collected on days 1 and 21 during maximal graded exercise and used for the determination of gross efficiency and whole body substrate use. Muscle biopsies were obtained before and after exercise on days 2 and 22 for the determination of mtDNA/gDNA ratio, gene expression, metabolic enzyme activity and glycogen use. Muscle glycogen before and after exercise, fat oxidation, and gross efficiency increased, carbohydrate oxidation decreased (p < 0.05), and VO(2max) did not change over the 21 days of training. Citrate synthase (CS), β-hydroxyacyl CoA dehydrogenase (β-HAD) and cytochrome c oxidase (COX) enzyme activity did not change with training. CS and β-HAD mRNA did not change with acute exercise or training. COX (subunit IV) mRNA increased with acute exercise (p < 0.05) but did not change over the 21 days. PGC-1α mRNA increased with acute exercise, but did not increase to the same degree on day 22 as it did on day 2 (p < 0.05). UCP3 mRNA decreased with training (p < 0.05). Acute exercise caused an increase in mitofusin2 (MFN2) mRNA (p < 0.05) and a trend for an increase in mtDNA/gDNA ratio (p = 0.057). However, training did not affect MFN2 mRNA or mtDNA/gDNA ratio. In response to 3,211 km of cycling, changes in substrate use and gross efficiency appear to be more profound than mitochondrial adaptations in trained individuals.

The aim of the present investigation was to examine the response of 80 plasma inflammatory analytes during five days of exercise in a hot (38 °C, 40% relative humidity) environment. 15 male participants (25±4yrs, 54±6 ml kg −1 min −1 VO 2 max), with no heat exposure within the previous 3 weeks, were asked to cycle in a hot environment at 70% of their VO 2 max workload until their terminal temperature was obtained, for 5 consecutive days. Terminal temperature was determined as the core temperature at volitional exhaustion or a core temperature of 39.5 °C, whichever came first. Blood samples were collected pre- and post-exercise on day 1 and day 5. Pre-trial urine specific gravity and body weight was not different on day 1 and day 5. Exercise time and heart rate at terminal temperature did not change during the five days. Of the 52 plasma analytes that increased in concentration on day 1, only 30 demonstrated increased concentrations at terminal temperature on day 5. Resting concentrations of 18, both pro- (IL-12p40, IL-15) and anti-inflammatory (IL-1ra, IL-10, IL-13) analytes were elevated on day 5 compared to day 1. We conclude that individuals completing consecutive days of exercise in the heat, but not definitively attaining heat acclimation, have increased resting levels of many inflammatory analytes associated with heat illness, but also demonstrate a reduced inflammatory response to a subsequent bout of exercise in the heat.

The purpose of this case study is to examine the physiological/behavioral factors leading up to heat exhaustion in a male wildland firefighter during wildland fire suppression. The participant (24 years old, 173 cm, 70 kg, and 3 years firefighting experience) experienced heat exhaustion following 7 hours of high ambient temperatures and arduous work on the fire line during the month of August. At the time of the heat-related incident (HRI), core temperature was 40.1 °C (104.2 °F) and skin temperature was 34.4 °C (93.9 °F). His work output averaged 1067 counts·min(-1) (arbitrary units for measuring activity) for the 7 hours prior to the HRI, a very high rate of work over an extended time period during wildfire suppression. In the 2.5 hours leading up to the heat incident, he was exposed to a mean ambient temperature of 44.6 °C (112.3 °F), with a maximum temperature of 59.7 °C (139.5 °F). He consumed an average of 840 mL·h(-1) in the 7 hours leading up to the incident and took an average of 24 ± 11 drinks·h(-1) (total of 170 drinks). The combined effects of a high work rate and high ambient temperatures resulted in an elevated core temperature and a higher volume and frequency of drinking than typically seen in this population, ultimately ending in heat exhaustion and removal from the fire line. The data demonstrate that heat-related incidents can occur even with aggressive fluid intake during wildland fire suppression.

Purpose:The purpose of this study was to determine the metabolic profile during the 2006 Ironman World Championship in Kailua-Kona, Hawaii.Methods:One recreational male triathlete completed the race in 10:40:16. Before the race, linear regression models were established from both laboratory and feld measures to estimate energy expenditure and substrate utilization. The subject was provided with an oral dose of 2H218O approximately 64 h before the race to calculate total energy expenditure (TEE) and water turnover with the doubly labeled water (DLW) technique. Body weight, blood sodium and hematocrit, and muscle glycogen (via muscle biopsy) were analyzed pre- and postrace.Results:The TEE from DLW and indirect calorimetry was similar: 37.3 MJ (8,926 kcal) and 37.8 MJ (9,029 kcal), respectively. Total body water turnover was 16.6 L, and body weight decreased 5.9 kg. Hematocrit increased from 46 to 51% PCV. Muscle glycogen decreased from 152 to 48 mmoL/kg wet weight pre- to postrace.Conclusion:These data demonstrate the unique physiological demands of the Ironman World Championship and should be considered by athletes and coaches to prepare sufficient nutritional and hydration plans.

The purpose of this study was to impose a period of quantifiable intensified training to determine if commonly used diagnostic markers of overtraining parallel changes in physical performance and thus overtraining status. Eight trained male cyclists (24 ± 1 years, 71 ± 3 kg, VO2peak = 4.5 ± 0.1·L·min⁻¹) performed 21 days (3,211 km) of intensified training in the field where volume and intensity were increased over normal training. Salivary IgA, testosterone, and cortisol, 1-hour time trial performance, heart rate response, and profile of mood states (POMS) were collected and analyzed throughout the 21-day training period. The POMS category vigor declined from day 1 to day 4 and remained lower throughout (p0.05). There were no other statistical changes in overtraining parameters. However, individuals who demonstrated 2 or more symptoms of overtraining at any point throughout the 21 days were considered symptomatic and had a lower (p0.05) VO2peak (4.2 ± 0.1·vs. 4.7 ± 0.1 L·min⁻¹) and lower (p0.05) average workload during the initial 1-hour time trial (253 ± 5 vs. 288 ± 14 W). Interestingly, the 1-hour time trial power in these individuals with symptoms of overtraining did not decline (p0.05). These data demonstrate that markers of overtraining do not parallel a decrease in performance and should be interpreted with caution.

The purpose of this study was to determine the effects of a recovery beverage immediately after exercise on rates of muscle glycogen resynthesis in response to road cycling when nutritional supplementation was supplied during exercise and a solid meal was served two hours after exercise. Eight trained male cyclists, (25+/-4 years, 69.3+/-5.2 kg, VO2 peak=4.5+/-0.4 L.min(-1)) performed two 62 km outdoor training rides in a double-blind, randomized cross-over experiment. Subjects received a food bar and a commercial sport drink during each ride. A recovery beverage (40 g CHO+20 g PRO) or a placebo (PL) was administered 30 min post-exercise. At 2 h post-exercise, a solid meal was provided for both trials. There was no difference between trials at any time point for glycogen (140+/-9, 56+/-8, and 70+/-8 mmol.kg(-1)wet wt.(-1).hr.(-1) for pre, post, and 4 h post, respectively). The addition of a supplemental recovery beverage ingested soon after exercise did not significantly increase the rate of muscle glycogen resynthesis after 4 h of recovery when nutritional supplementation is provided during exercise and a meal is consumed 2 h after exercise.