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Parasitic nematodes (roundworms) of small ruminants and other livestock have major economic impacts worldwide. Despite the impact of the diseases caused by these nematodes and the discovery of new therapeutic agents (anthelmintics), there has been relatively limited progress in the development of practical molecular tools to study the epidemiology of these nematodes. Specific diagnosis underpins parasite control, and the detection and monitoring of anthelmintic resistance in livestock parasites, presently a major concern around the world. The purpose of the present article is to provide a concise account of the biology and knowledge of the epidemiology of the gastrointestinal nematodes (order Strongylida), from an Australian perspective, and to emphasize the importance of utilizing advanced molecular tools for the specific diagnosis of nematode infections for refined investigations of parasite epidemiology and drug resistance detection in combination with conventional methods. It also gives a perspective on the possibility of harnessing genetic, genomic and bioinformatic technologies to better understand parasites and control parasitic diseases.

Background A lack of safety and dosing information on restoring hormone levels in neutered dogs has hindered treatment of spay-neuter syndrome, the diverse set of health and behavior conditions resulting from the loss of gonadal hormones. The purpose of this study was to provide basic information on the use of testosterone treatment in male neutered dogs using a target animal safety study design. Twelve previously neutered dogs were divided into four equal groups, receiving 0 (controls), 1x, 3x, or 5x the standard weekly dose of injectable testosterone cypionate (0.5 mg/kg) for 90 days. Bloodwork, including endocrine assays, body condition, prostate health, and qualitative behavioral observations were used to evaluate any changes in health during treatment. Results Testosterone levels increased in a dose-dependent fashion, and luteinizing hormone decreased after a month of treatment in dogs receiving 5x the standard dose. Behavioral measures, Zambelli prostate health score, body condition scores, clinical evaluations, and routine blood hematology and chemistries showed minor variation over time or across groups. Two seizures were documented in a dog from group 3x with previous suspected idiopathic epilepsy. Conclusions The results of the initial safety study indicated that weekly injectable testosterone therapy over a 3-month period was safe and capable of increasing testosterone levels in neutered dogs to within the normal range for intact dogs. The limitations of the study included small group sizes and no long-term follow up. A summary of the risks and recommendations for the use of testosterone restoration in dogs was provided.

Background Traditional methods of detecting and identifying gastrointestinal nematode infections in small ruminants, including sheep and goats, are time-consuming and lack in sensitivity and specificity. Recently, we developed an automated multiplexed-tandem (MT)-PCR platform for the diagnosis and identification patent infections with key genera/species of gastrointestinal nematodes of sheep and validated this approach in detailed experiments carried out in Australia. In the present study, we deployed this diagnostic platform in Scotland and Belgium to test samples from naturally infected sheep in these environments and to validate the MT-PCR platform relative to traditional diagnostic methods routinely used by local laboratories. Results MT-PCR detected all microscopy positive samples and there was a significant agreement between the results of the different test methods in terms of the species detected and their relative proportion in a test sample, however, for some samples, there were discrepancies between the results of the different test methods. Selective sequencing of purified MT-PCR products demonstrated the results to be 100% specific. Conclusions The MT-PCR platform is an advanced method for the species/genus-specific diagnosis of gastrointestinal nematode infections in small ruminants and has demonstrated utility when deployed in different countries and climatic zones. The platform is user-friendly due to the largely automated procedure and has high versatility in that it can achieve a specific diagnosis from different types of sample templates, including larval culture and faecal samples. With appropriate modifications of the primers used, the MT-PCR platform also provides potential for the diagnosis of a variety of other pathogens of veterinary or medical importance.

The accurate diagnosis of parasitic nematode infections in livestock (including sheep and goats) is central to their effective control and the detection of the anthelmintic resistance. Traditionally, the faecal egg count reduction test (FECRT), combined with the technique of larval culture (LC), has been used widely to assess drug-susceptibility/resistance in strongylid nematodes. However, this approach suffers from a lack of specificity, sensitivity and reliability, and is time-consuming and costly to conduct. Here, we critically assessed a specific PCR assay to support FECRT, in a well-controlled experiment on sheep with naturally acquired strongylid infections known to be resistant to benzimidazoles. We showed that the PCR results were in close agreement with those of total worm count (TWC), but not of LC. Importantly, albendazole resistance detected by PCR-coupled FECRT was unequivocally linked to Teladorsagia circumcincta and, to lesser extent, Trichostrongylus colubriformis, a result that was not achievable by LC. The key findings from this study demonstrate that our PCR-coupled FECRT approach has major merit for supporting anthelmintic resistance in nematode populations. The findings also show clearly that our PCR assay can be used as an alternative to LC, and is more time-efficient and less laborious, which has important practical implications for the effective management and control strongylid nematodes of sheep.

In roots of Arabidopsis (Arabidopsis thaliana), l-lactate is generated by the reduction of pyruvate via l-lactate dehydrogenase, but this enzyme does not efficiently catalyze the reverse reaction. Here, we identify the Arabidopsis glycolate oxidase (GOX) paralogs GOX1, GOX2, and GOX3 as putative l-lactate-metabolizing enzymes based on their homology to CYB2, the l-lactate cytochrome c oxidoreductase from the yeast Saccharomyces cerevisiae. We found that GOX3 uses l-lactate with a similar efficiency to glycolate; in contrast, the photorespiratory isoforms GOX1 and GOX2, which share similar enzymatic properties, use glycolate with much higher efficiencies than l-lactate. The key factor making GOX3 more efficient with l-lactate than GOX1 and GOX2 is a 5- to 10-fold lower Km for the substrate. Consequently, only GOX3 can efficiently metabolize l-lactate at low intracellular concentrations. Isotope tracer experiments as well as substrate toxicity tests using GOX3 loss-of-function and overexpressor plants indicate that l-lactate is metabolized in vivo by GOX3. Moreover, GOX3 rescues the lethal growth phenotype of a yeast strain lacking CYB2, which cannot grow on l-lactate as a sole carbon source. GOX3 is predominantly present in roots and mature to aging leaves but is largely absent from young photosynthetic leaves, indicating that it plays a role predominantly in heterotrophic rather than autotrophic tissues, at least under standard growth conditions. In roots of plants grown under normoxic conditions, loss of function of GOX3 induces metabolic rearrangements that mirror wild-type responses under hypoxia. Thus, we identified GOX3 as the enzyme that metabolizes l-lactate to pyruvate in vivo and hypothesize that it may ensure the sustainment of low levels of l-lactate after its formation under normoxia.

Coccolithophores have influenced the global climate for over 200 million years1. These marine phytoplankton can account for 20 per cent of total carbon fixation in some systems2. They form blooms that can occupy hundreds of thousands of square kilometres and are distinguished by their elegantly sculpted calcium carbonate exoskeletons (coccoliths), rendering them visible from space3. Although coccolithophores export carbon in the form of organic matter and calcite to the sea floor, they also release CO2 in the calcification process. Hence, they have a complex influence on the carbon cycle, driving either CO2 production or uptake, sequestration and export to the deep ocean4. Here we report the first haptophyte reference genome, from the coccolithophore Emiliania huxleyi strain CCMP1516, and sequences from 13 additional isolates. Our analyses reveal a pan genome (core genes plus genes distributed variably between strains) probably supported by an atypical complement of repetitive sequence in the genome. Comparisons across strains demonstrate that E. huxleyi, which has long been considered a single species, harbours extensive genome variability reflected in different metabolic repertoires. Genome variability within this species complex seems to underpin its capacity both to thrive in habitats ranging from the equator to the subarctic and to form large-scale episodic blooms under a wide variety of environmental conditions.

In roots of Arabidopsis (Arabidopsis thaliana), L-lactate is generated by the reduction of pyruvate via L-lactate dehydrogenase, but this enzyme does not efficiently catalyze the reverse reaction. Here, we identify the Arabidopsis glycolate oxidase (GOX) paralogs GOX1, GOX2, and GOX3 as putative L-lactate-metabolizing enzymes based on their homology to CYB2, the L-lactate cytochromecoxidoreductase from the yeastSaccharomyces cerevisiae. We found that GOX3 uses L-lactate with a similar efficiency to glycolate; in contrast, the photorespiratory isoforms GOX1 and GOX2, which share similar enzymatic properties, use glycolate with much higher efficiencies than L-lactate. The key factor making GOX3 more efficient with L-lactate than GOX1 and GOX2 is a 5- to 10-fold lowerK mfor the substrate. Consequently, only GOX3 can efficiently metabolize L-lactate at low intracellular concentrations. Isotope tracer experiments as well as substrate toxicity tests usingGOX3loss-offunction and overexpressor plants indicate that L-lactate is metabolized in vivo by GOX3. Moreover,GOX3rescues the lethal growth phenotype of a yeast strain lacking CYB2, which cannot grow on L-lactate as a sole carbon source. GOX3 is predominantly present in roots and mature to aging leaves but is largely absent from young photosynthetic leaves, indicating that it plays a role predominantly in heterotrophic rather than autotrophic tissues, at least under standard growth conditions. In roots of plants grown under normoxic conditions, loss of function ofGOX3induces metabolic rearrangements that mirror wild-type responses under hypoxia. Thus, we identified GOX3 as the enzyme that metabolizes L-lactate to pyruvate in vivo and hypothesize that it may ensure the sustainment of low levels of L-lactate after its formation under normoxia.

To enhance understanding of Earth's climate, numerical experiments are performed contrasting a retrograde and prograde rotating Earth using the Max Planck Institute Earth system model. The experiments show that the sense of rotation has relatively little impact on the globally and zonally averaged energy budgets but leads to large shifts in continental climates, patterns of precipitation, and regions of deep water formation. Changes in the zonal asymmetries of the continental climates are expected given ideas developed more than a hundred years ago. Unexpected was, however, the switch in the character of the European–African climate with that of the Americas, with a drying of the former and a greening of the latter. Also unexpected was a shift in the storm track activity from the oceans to the land in the Northern Hemisphere. The different patterns of storms and changes in the direction of the trades influence fresh water transport, which may underpin the change of the role of the North Atlantic and the Pacific in terms of deep water formation, overturning and northward oceanic heat transport. These changes greatly influence northern hemispheric climate and atmospheric heat transport by eddies in ways that appear energetically consistent with a southward shift of the zonally and annually averaged tropical rain bands. Differences between the zonally averaged energy budget and the rain band shifts leave the door open, however, for an important role for stationary eddies in determining the position of tropical rains. Changes in ocean biogeochemistry largely follow shifts in ocean circulation, but the emergence of a “super” oxygen minimum zone in the Indian Ocean is not expected. The upwelling of phosphate-enriched and nitrate-depleted water provokes a dominance of cyanobacteria over bulk phytoplankton over vast areas – a phenomenon not observed in the prograde model. What would the climate of Earth look like if it would rotate in the reversed (retrograde) direction? Which of the characteristic climate patterns in the ocean, atmosphere, or land that are observed in a present-day climate are the result of the direction of Earth's rotation? Is, for example, the structure of the oceanic meridional overturning circulation (MOC) a consequence of the interplay of basin location and rotation direction? In experiments with the Max Planck Institute Earth system model (MPI-ESM), we investigate the effects of a retrograde rotation in all aspects of the climate system. The expected consequences of a retrograde rotation are reversals of the zonal wind and ocean circulation patterns. These changes are associated with major shifts in the temperature and precipitation patterns. For example, the temperature gradient between Europe and eastern Siberia is reversed, and the Sahara greens, while large parts of the Americas become deserts. Interestingly, the Intertropical Convergence Zone (ITCZ) shifts southward and the modeled double ITCZ in the Pacific changes to a single ITCZ, a result of zonal asymmetries in the structure of the tropical circulation. One of the most prominent non-trivial effects of a retrograde rotation is a collapse of the Atlantic MOC, while a strong overturning cell emerges in the Pacific. This clearly shows that the position of the MOC is not controlled by the sizes of the basins or by mountain chains splitting the continents in unequal runoff basins but by the location of the basins relative to the dominant wind directions. As a consequence of the changes in the ocean circulation, a “super” oxygen minimum zone develops in the Indian Ocean leading to upwelling of phosphate-enriched and nitrate-depleted water. These conditions provoke a dominance of cyanobacteria over bulk phytoplankton over vast areas, a phenomenon not observed in the prograde model.