The Root-Lesion Nematode, Pratylenchus penetrans, Affects Early Growth and Physiology of Grafted M.9, G.41, and G.935 Apple Rootstocks Similarly Under Field Microplot Conditions.

The root-lesion nematode, Pratylenchus penetrans, is a ubiquitous parasite of roots of temperate fruit trees. It affects early growth of trees replanted into former orchard sites where populations have built up and may contribute to decline complexes of older trees. Most British Columbia, Canada, apple acreage is planted with M.9 rootstock, but growers are increasingly considering Geneva-series rootstocks such as G.41 and G.935. Among these rootstocks, responses to P. penetrans, specifically, are poorly known. To compare the resistance and tolerance to P. penetrans of G.41, G.935, and M.9 rootstocks ('Ambrosia' scion), a field microplot experiment was established in spring of 2020 at the Summerland Research and Development Centre. The experimental design was a two by three factorial combination of: P. penetrans inoculation (+/-) and rootstock (G.41, G.935, and M.9), with 20 replicate microplots of each of the six treatment combinations arranged in a randomized complete block design. The P. penetrans inoculum was 5,400 nematodes per microplot (54 P. penetrans liter-1 soil), which is below commonly accepted damage thresholds. Though P. penetrans population densities were lower for the G.41 rootstock by the end of the 2021 growing season, the effects of P. penetrans were similar among rootstocks. In the establishment year (2020), P. penetrans caused significant reductions in aboveground growth. In 2021, shoot growth and root weight were reduced by P. penetrans. The nematode also reduced rates of leaf gas exchange and stem water potential. These data suggest that while G.41 and G.935 may have other horticultural benefits over M.9, they are equally susceptible to P. penetrans at the early stages of tree growth.

Pseudomonads contribute to regulation of Pratylenchus penetrans (Nematoda) populations on apple.

Inoculation with antagonistic soil microorganisms has shown potential to suppress replant disease of apple in orchard soils. Pseudomonas spp. may have the potential to reduce Pratylenchus penetrans populations on apple. Pseudomonas spp. were isolated from the rhizosphere of sweet cherry and screened for antagonistic characteristics. Two highly antagonistic Pseudomonas isolates, P10-32 and P10-42, were evaluated for growth promotion of apple seedlings, suppression of P. penetrans populations, and root colonization in soil from three orchards. During the isolate screening, Pseudomonas fluorescens P10-32 reduced in vitro growth of fungal pathogens, had protease activity, had capacity to produce pyrrolnitrin, suppressed P. penetrans populations, and increased plant biomass. Pseudomonas fluorescens P10-42 reduced in vitro growth of fungal pathogens, had protease activity, suppressed P. penetrans populations, and increased plant biomass. In potted orchard soil, inoculating apple with P. fluorescens P10-32 suppressed P. penetrans populations in one of the three soils examined. Inoculation with P. fluorescens P10-42 improved plant growth in two of the soils and suppressed P. penetrans abundance in one soil. In one of the soils, P. fluorescens P10-42 was detected on the roots 56 days postinoculation. Overall, we conclude that Pseudomonas spp. play a role in suppressing P. penetrans on apple in orchard soil.

Using C. elegans Forward and Reverse Genetics to Identify New Compounds with Anthelmintic Activity.

BACKGROUND: The lack of new anthelmintic agents is of growing concern because it affects human health and our food supply, as both livestock and plants are affected. Two principal factors contribute to this problem. First, nematode resistance to anthelmintic drugs is increasing worldwide and second, many effective nematicides pose environmental hazards. In this paper we address this problem by deploying a high throughput screening platform for anthelmintic drug discovery using the nematode Caenorhabditis elegans as a surrogate for infectious nematodes. This method offers the possibility of identifying new anthelmintics in a cost-effective and timely manner. METHODS/PRINCIPAL FINDINGS: Using our high throughput screening platform we have identified 14 new potential anthelmintics by screening more than 26,000 compounds from the Chembridge and Maybridge chemical libraries. Using phylogenetic profiling we identified a subset of the 14 compounds as potential anthelmintics based on the relative sensitivity of C. elegans when compared to yeast and mammalian cells in culture. We showed that a subset of these compounds might employ mechanisms distinct from currently used anthelmintics by testing diverse drug resistant strains of C. elegans. One of these newly identified compounds targets mitochondrial complex II, and we used structural analysis of the target to suggest how differential binding of this compound may account for its different effects in nematodes versus mammalian cells. CONCLUSIONS/SIGNIFICANCE: The challenge of anthelmintic drug discovery is exacerbated by several factors; including, 1) the biochemical similarity between host and parasite genomes, 2) the geographic location of parasitic nematodes and 3) the rapid development of resistance. Accordingly, an approach that can screen large compound collections rapidly is required. C. elegans as a surrogate parasite offers the ability to screen compounds rapidly and, equally importantly, with specificity, thus reducing the potential toxicity of these compounds to the host and the environment. We believe this approach will help to replenish the pipeline of potential nematicides.

Soil Aggregate Dynamics and Plant Community Response after Biosolids Application in a Semiarid Grassland.

Biosolids may improve the ecological function of degraded semiarid grasslands, but an understanding of the plant community is essential. An experiment was established in 2001 to determine the effects of a single surface application of biosolids on soil aggregate stability and the composition of the plant community in a semiarid grassland in British Columbia, Canada. Four treatments were evaluated: (i) surface biosolids application at 20 (Bio-20) and (ii) 60 Mg ha (Bio-60), (iii) mineral fertilizer, and (iv) a control. All treatments were replicated in four blocks. Soil was sampled during the spring (May), summer (June-July), and fall (October) in 2005, 2006, and 2009; the plant community was assessed in 2009. The greatest increases in size of stable aggregates relative to the control were in the spring and summer, which coincided with a 1.6- to 2.1-fold increase in the spring concentration of N within stable aggregates when biosolids were applied at 20 and 60 Mg ha , respectively. Nitrogen concentrations from the Bio-60 treatment were not different from the control, but the Bio-20 treatment had 42% greater N than all other treatments during summer. Biosolids application in this ecosystem did not increase perennial forage grass species relative to the control, and when biosolids were applied at a rate of 60 Mg ha there was a 75% reduction in the perennial forage plant species. The application of biosolids to native grasslands in semiarid environments should be done cautiously, especially when winter annual plant species (e.g., cheatgrass [ L.]) are present before application.

Characterization of antibiotic-resistant and potentially pathogenic Escherichia coli from soil fertilized with litter of broiler chickens fed antimicrobial-supplemented diets.

The objective of this study was to characterize antimicrobial resistance and virulence determinants of Escherichia coli from soil amended with litter from 36-day-old broiler chickens ( Gallus gallus domesticus ) fed with diets supplemented with a variety of antimicrobial agents. Soil samples were collected from plots before and periodically after litter application in August to measure E. coli numbers. A total of 295 E. coli were isolated from fertilized soil samples between August and March. Antibiotic susceptibility was determined by Sensititre, and polymerase chain reaction was performed to detect the presence of resistance and virulence genes. The results confirmed that E. coli survived and could be quantified by direct plate count for at least 7 months in soil following litter application in August. The effects of feed supplementation were observed on E. coli numbers in November and January. Among the 295 E. coli, the highest antibiotic resistance level was observed against tetracycline and ß-lactams associated mainly with the resistance genes tetB and bla(CMY-2), respectively. Significant treatment effects were observed for phylogenetic groups, antibiotic resistance profiles, and virulence gene frequencies. Serotyping, phylogenetic grouping, and pulsed-field gel electrophoresis confirmed that multiple-antibiotic-resistant and potentially pathogenic E. coli can survive in soil fertilized with litter for several months regardless of antimicrobials used in the feed.

Biosolids increase soil aggregation and protection of soil carbon five years after application on a crested wheatgrass pasture.

Biosolids application to rangelands and pastures recycles nutrients and organic matter back to soils. The effects of biosolids (20 and 60 dry Mg ha(-)(1)) and N+P fertilizer on soil aggregate stability, bulk density, aeration porosity, and total C and N of stable aggregates were evaluated 4 and 5 yr after surface application to a crested wheatgrass [Agropyron cristatum (L.) Gaertn.] pasture in the southern interior of British Columbia (BC). The experiment was established in 2001 in a randomized complete block design with four replications. The 60 Mg ha(-1) biosolids treatment (Bio 60) had a greater aggregate mean weight diameter (MWD) and proportion of water-stable soil aggregates > 1 mm relative to the control and fertilizer treatments. Temporal variation in aggregate stability was attributed to seasonal variations in soil water content. Surface application of 60 Mg ha(-1) of biosolids increased C concentrations within water-stable aggregates relative to the control from 29 to 104, 24 to 79, and 12 to 38 g kg(-1) for the 2 to 6, 1 to 2, and 0.25 to 1 mm size fractions, respectively. The concentration of N within aggregates increased in similar proportions to C. Neither soil bulk density, nor aeration porosity were affected by biosolids application. Increased aggregation and the accumulation of soil C within aggregates following biosolids application creates a potential for better soil C storage, soil water retention, nutrient availability, and ultimately the overall health of semiarid perennial pastures.