Differential Metal Binding Interactions of the Imidazolinones Revealed by NMR and UV Spectroscopy

NMR and UV spectroscopy and molecular modeling methods were applied to probe the interaction of the two imidazolinones, imazethapyr (5-ethyl-2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)nicotinic acid) and its structural isomer CL 303,135 (5-ethyl-3-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)picolinic acid), with metal ions. Both the imidazolinones inhibit the enzyme acetohydroxyacid synthase (AHAS) in vitro. However, while imazethapyr is a herbicide that is used widely in agriculture, CL303,135 does not exhibit herbicidal activity. Imazethapyr and CL303,135 exhibited considerable differences in their interactions with metals. In the metal complex of imazethapyr, the carboxyl moiety binds strongly and the pyridine nitrogen binds weakly with metals. In the case of CL303,135, both the pyridine nitrogen and the carboxyl group that are positioned ortho to each other participated strongly in the binding and were found to act together as a strong bidentate ligand to a metal ion. Both of the imidazolinones form predominantly 2:1 complexes with multivalent metal ions. However, imazethapyr binds two orders-of-magnitude more weakly (1·0×10⁹ M ^-2) with metal ions compared to CL303,135 (1·7×10¹¹ M ^-2). The interactions of the model compounds, nicotinic acid and picolinic acid, with metals were examined similarly. It was concluded that the strong affinity of CL303,135 for metals compared to imazethapyr may affect its absorption from soil into plants, or its translocation in plants, thereby explaining the differences in herbicidal activity of imazethapyr and CL303,135. © 1997 SCI.     

Contrasting soil microbial responses to fertilizers and herbicides in a canola-barley rotation

The combination of high input costs and low commodity prices is forcing some farmers to consider reducing crop inputs like seed, fertilizer and herbicides. In a field trial in which different canola (Brassica napus L.) and barley (Hordeum vulgare L.) inputs were subtracted from a full package, or added to an empty package, we studied the effects of full or reduced fertilizer and herbicide inputs on soil microbiological characteristics at two sites from 2005 to 2008. The full package consisted of a high-yielding crop variety seeded at an optimum rate, with fertilizers and herbicides applied at recommended rates. The empty package consisted of a less expensive, low-yielding crop variety seeded at a low rate, with no fertilizer or herbicide applied. Between these two extremes were treatments in which fertilizers or herbicides were applied at 50% of recommended rates or not at all. Each treatment was repeated year after year in the same plot, i.e., treatment effects were cumulative. Fertilizer effects on soil microbial biomass C (MBC), β-glucosidase enzyme activity and bacterial functional diversity (based on community-level physiological profiles) were usually positive. Reduced fertilizer application rates reduced the beneficial fertilizer effects. Significant herbicide effects on soil microbiological properties occurred less often, were smaller in magnitude than fertilizer effects, and were mostly negative. Reduced herbicide rates reduced the deleterious herbicide effects. These significant fertilizer and herbicide effects were observed in canola more than barley, and mostly in the final year of the study, indicating the cumulative nature of treatment effects over time. Therefore, repeated applications of agricultural inputs like fertilizers and herbicides can have more significant effects on soil biology and biological processes than single applications indicate.     

Early Warnings for State Transitions

New concepts have emerged in theoretical ecology with the intent to quantify complexities in ecological change that are unaccounted for in state-and-transition models and to provide applied ecologists with statistical early warning metrics able to predict and prevent state transitions. With its rich history of furthering ecological theory and its robust and broad-scale monitoring frameworks, the rangeland discipline is poised to empirically assess these newly proposed ideas while also serving as early adopters of novel statistical metrics that provide advanced warning of a pending shift to an alternative ecological regime. We review multivariate early warning and regime shift detection metrics, identify situations where various metrics will be most useful for rangeland science, and then highlight known shortcomings. Our review of a suite of multivariate-based regime shift/early warning indicators provides a broad range of metrics applicable to a wide variety of data types or contexts, from situations where a great deal is known about the key system drivers and a regime shift is hypothesized a priori, to situations where the key drivers and the possibility of a regime shift are both unknown. These metrics can be used to answer ecological state-and-transition questions, inform policymakers, and provide quantitative decision-making tools for managers.     

Genotype Rankings for Nutrient Stress Resistance are Unrelated to Stress Severity in Cultivated Sunflower (Helianthus annuus L.)

Abiotic stress limits crop productivity worldwide, making stress‐resistant crops a major goal for improving food security. Stress resistance is often assessed by comparing genotypes under a single control and stressed level of an abiotic factor, then ranking the genotypes’ overall performance, using a metric like geometric mean productivity (GMP). However, abiotic stress is rarely a presence/absence phenomenon, but often varies continuously. Here, we tested whether the common practice of assessing performance using a single control and stress level is predictive of performance across a range of stress levels. Twelve sunflower genotypes were grown in the greenhouse under one of eight nutrient levels (from non‐limiting to severely limiting) and assessed for performance (shoot dry mass at the budding stage) and related traits at both seedling and budding stages. Contrary to expectations, genotype rankings for GMP were consistent regardless of the two nutrient levels used to calculate GMP. This suggests that, for nutrient stress in sunflower, studies using an arbitrary control and stress level are broadly representative: genotypes with high GMP exhibit high GMP regardless of the nutrient levels used. In addition, GMP values were highly correlated with performance in the respective nutrient levels from which they were calculated; thus, higher GMP for a given genotype indicates that it performed well (relative to other genotypes) under both stressed and non‐stressed conditions. Together, these findings strongly support the current paradigm of comparing genotypes at a single control and stress level as a valid strategy for screening for nutrient stress resistance in sunflower. However, few traits were consistently associated with performance (shoot dry mass) at either high‐ or low‐nutrient availability, and seedling traits were rarely predictive of budding‐stage traits and performance, suggesting further studies are needed to determine the mechanisms underlying nutrient stress resistance in this important oilseed crop.     

Quantifying home range habitat requirements for bobcats (Lynx rufus) in Vermont,USA

We demonstrate how home range and habitat use analysis can inform landscape-scale conservation planning for the bobcat, Lynx rufus, in Vermont USA. From 2005 to 2008, we outfitted fourteen bobcats with GPS collars that collected spatially explicit locations from individuals every 4 h for 3–4 months. Kernel home range techniques were used to estimate home range size and boundaries, and to quantify the utilization distribution (UD), which is a spatially explicit, topographic mapping of how different areas within the home range are used. We then used GIS methods to quantify both biotic (e.g. habitat types, stream density) and abiotic (e.g. slope) resources within each bobcat’s home range. Across bobcats, upper 20th UD percentiles (core areas) had 18% less agriculture, 42% less development, 26% more bobcat habitat (shrub, deciduous, coniferous forest, and wetland cover types), and 33% lower road density than lower UD percentiles (UD valleys). For each bobcat, we used Akaike’s Information Criterion (AIC) to evaluate and compare 24 alternative Resource Utilization Functions (hypotheses) that could explain the topology of the individual’s UD. A model-averaged population-level Resource Utilization Function suggested positive responses to shrub, deciduous, coniferous forest, and wetland cover types within 1 km of a location, and negative responses to roads and mixed forest cover types within 1 km of a location. Applying this model-averaged function to each pixel in the study area revealed habitat suitability for bobcats across the entire study area, with suitability scores ranging between ?1.69 and 1.44, where higher values were assumed to represent higher quality habitat. The southern Champlain Valley, which contained ample wetland and shrub habitat, was a concentrated area of highly suitable habitat, while areas at higher elevation areas were less suitable. Female bobcat home ranges, on average, had an average habitat suitability score of near 0, indicating that home ranges consisted of both beneficial and detrimental habitat types. We discuss the application of habitat suitability mapping and home range requirements for bobcat conservation and landscape scale management.     

The effect of various Mycoplasma bovis isolates on bovine leukocyte responses

Mycoplasma bovis (M. bovis) contributes to a number of clinical syndromes in cattle; in particular, chronic pneumonia that is poorly responsive to therapy has been increasingly recognized as an important cause of morbidity, mortality, and financial loss. M. bovis impairs host immune function, but little is known about whether field isolates vary significantly in their effect on immune function. This research tested the hypothesis that different field isolates vary in their ability to suppress cellular metabolism and cellular production of radical oxygen species (ROS) by bovine leukocytes. Total blood leukocytes from 6 cattle were exposed to six field isolates, two diagnostic lab isolates, and two high passage laboratory isolates of M. bovis, and ROS production was measured by oxidation of dihydrorhodamine 123 (DHR-123). Cellular metabolism was measured by reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Significant differences in the response to some field isolates were identified. Three field isolates and both diagnostic lab isolates significantly decreased ROS production by leukocytes from multiple cattle, while the high pass laboratory isolates did not. In contrast, MTT reduction was not significantly impaired by any of the M. bovis strains tested. M. bovis impairs ROS production by bovine leukocytes; the magnitude of the effect appears to be isolate-dependent, and is not related to a general impairment of cellular metabolism. Chronic M. bovis infection in some cattle may be related to impaired ability of leukocytes to produce ROS when exposed to M. bovis.     

Synthetic and mechanistic studies on fungal metabolic pathways: A guide to fungicide design

The polyketide biosynthetic pathway is responsible for the formation in microorganisms and plants of a vast array of diverse structures, many of which display important biological activity. A brief overview of the pathway, with emphasis on present problems and future developments, is presented and the impact of genetics on chemical and biochemical studies of polyketide biosynthesis is highlighted. Biosynthetic and mechanistic studies on three pathways are described, to illustrate how these studies may provide an insight into the mode of action of particular compounds, or how particular pathways may be inhibited. LL-D253α is an antibiotic chromanone produced by a number of Phoma species. Stable isotope labelling studies have indicated the involvement of cyclopropylcyclohexadienyl intermediates in the formation of the hydroxyethyl side chain indicating a possible mode of action. Similar studies of monocerin indicate the involvement of quinone methide intermediates. A synthesis of monocerin, modelled on the biosynthetic pathway, is described. Scytalone and vermelone are intermediates on the pathway to melanin in certain pathogenic fungi, e.g. Pyricularia oryzae. Progress with biosynthetic studies on this pathway is described.     

The relationship of training milestones with racing success in a population of Standardbred horses in New Zealand

AIM: To gather information on the repeatability of a faecal nematode egg count (FEC) reduction (FECR) test (FECRT), evaluating both different methods of calculating efficacy and variations within a method, in order to supply veterinarians and other advisors with sufficient information to apply some level of confidence around a diagnosis of anthelmintic resistance based on FECRT results.

METHODS: Two commercial sheep farms were selected on the basis of having previously recorded FECR <95% after treatment with ivermectin (Farm 1) or albendazole (Farm 2). On each farm at least 250 lambs, managed as a single mob, were individually ear-tagged and sampled for FEC. The resulting counts were used, 3—4 days later, to sort the lambs into 24 groups of 10. First, the animals were split into three groups of 80, having high, medium or low FEC. Second, within each of these groups the 80 animals were further divided into four replicate mobs of 20 (each with the same mean count). Third, each of these replicates was further split into two groups of 10: those that would be drenched and those that would remain as untreated controls. All animals were again faecal-sampled and those in the drenched groups were dosed, using a syringe, to their individual liveweight, with ivermectin (Farm 1) or albendazole (Farm 2). Ten days after treatment all animals were individually faecal- sampled again. FEC and larval cultures were undertaken for all 24 groups from both pre- and post-treatment samples. Ef- ficacy (FECR) of the undifferentiated FECRT was calculated using three different equations, and efficacy by genus was also calculated.

RESULTS: Calculated efficacies differed between equations, and the equation which did not utilise an untreated control yielded significantly lower efficacy estimates on both farms. Faecal cultures varied considerably in the proportions of parasite genera recovered. In general, this did not differ between FEC groups, except on Farm 1 where Haemonchus spp were more common and Cooperia spp less common in high-FEC samples. Estimated efficacies against individual genera varied considerably or very little, depending on the level of resistance. On both farms, differing proportions of tests against some genera passed or failed FECRTs based on a threshold pass mark of ≥95% FECR.

CONCLUSION: There was considerable variability in the outcomes of FECRTs and in larval culture results. Caution is warranted in interpreting the results of FECRTs when efficacy values fall into the 90—95% range. Further, the possibility of a test returning a false-negative result is raised, indicating that even an efficacy estimated ≥95% may not guarantee the absence of resistant parasites.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 1. Patterns of seasonal development

Abstract

AIM: To describe the seasonal pattern of development of third-stage infective larvae (L3) from eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition on 13–18 occasions, between January 2005 and July 2006. Analysis of the number of larvae recovered used a mixed model including number of eggs deposited, weight of faeces recovered (an assumed indicator of earthworm activity), site, contamination date, and position of deposited faeces, i.e. on the surface or buried.

RESULTS: There was a significant effect of contamination date on development of all three species, with maximum numbers ofL3 developing between late spring (November) and early autumn (March), and minimum numbers in June and July. There were large differences between species, with H. contortus exhibiting a long period (April to October) where development was close to zero, whereas T. circumcincta developed to some extent all year round. Development of T. colubriformis was intermediate between the other two species.

Burying faeces containing nematode eggs increased the number of L3 recovered compared with surface deposition (p≤0.001), although there were a small number of exceptions involving only T. colubriformis. The weight of faeces recovered at harvest, which was assumed to be an indication of earthworm activity, was correlated with the number of L3 recovered for all species (p<0.001). In a separate analysis, earthworms were assumed tohave been active if <5 g faeces remained at harvest. Where this occurred, the number of L3 of T. colubriformis and T.circumcincta recovered was reduced by 56% and 58%, respectively (p<0.001).

CONCLUSIONS: A marked seasonal pattern of development was observed for all three species, with the most larvae developing in spring-early autumn and the least in winter. This seasonal pattern was most pronounced in H. contortus and least obvious in T. circumcincta. Burying faeces containing eggs generally resulted in more L3 being recovered, whilst the apparent activity of earthworms resulted in fewer larvae being recovered.