Grass-Endophyte Interactions and Their Associated Alkaloids as a Potential Management Strategy for Plant Parasitic Nematodes.

Claviceptaceous endophytic fungi in the genus Epichloë mostly form a symbiotic relationship with cool-season grasses. Epichloë spp. are capable of producing bioactive alkaloids such as peramines, lolines, ergot alkaloids, and indole-diterpenes, which protect the host plant from herbivory by animals, insects, and nematodes. The host also benefits from enhanced tolerance to abiotic stresses, such as salt, drought, waterlogging, cold, heavy metals, and low nitrogen stress. The bioactive alkaloids produced can have both direct and indirect effects towards plant parasitic nematodes. Direct interaction with nematodes' motile stages can cause paralysis (nematostatic effect) or death (nematicidal effect). Indirectly, the metabolites may induce host immunity which inhibits feeding and subsequent nematode development. This review highlights the different mechanisms through which this interaction and the metabolites produced have been explored in the suppression of plant parasitic nematodes and also how the specific interactions between different grass genotypes and endophyte strains result in variable suppression of different nematode species. An understanding of the different grass-endophyte interactions and their successes and failures in suppressing various nematode species is essential to enable the proper selection of grass-endophyte combinations to identify the alkaloids produced, concentrations required, and determine which nematodes are sensitive to which specific alkaloids.

Screening of Fungicides and Comparison of Selective Media for Isolation of Fusarium graminearum from Soil and Plant Material.

The culture media recommended for the isolation and enumeration of the Fusarium spp. lack selectivity for Fusarium graminearum. Five fungicides-Amistar® (250 g·L-1 azoxystrobin), Filan® (500 g·kg-1 boscalid), Comet® 200 (200 g·L-1 pyraclostrobin), Imtrex® (62.5 g·L-1 fluxapyroxad), Poraz® (450 g·L-1 prochloraz)-were investigated for their potential as selective inhibitors in culture media for the isolation of F. graminearum from soil and plant material. Based on the screening, fluxapyroxad was further tested for selective inhibition for the isolation of F. graminearum from soil. Additionally, selective media were compared for the isolation of F. graminearum from plant material. The fungicides tested did not prove to be effective inhibitors for the development of selective media. For the detection of F. graminearum in plant material, Czapek Dox propiconazole dichloran agar was found to be a better medium than Komada's media, as the former resulted in colonies with darker pigmentation over a shorter incubation time and appeared to have a less inhibitory effect on F. graminearum growth.

Biofumigation for the Management of Fusarium graminearum in a Wheat-Maize Rotation.

Fusarium graminearum is the most important causal agent of head blight in wheat, and stalk and ear rot in maize. A field experiment was conducted to investigate the effect of incorporation of Brassicaceae cover crops on Fusarium graminearum in a wheat-maize rotation. Five species belonging to Brassicaceae (Brassica juncea, Eruca sativa, Raphanus sativus, B. carinata, B. oleracea var. caulorapa L.) were used in the field experiment to investigate their potential to suppress F. graminearum inoculum in soil, disease incidence in maize and to reduce subsequent mycotoxin contamination in maize. Brassica juncea was found to contain the highest glucosinolate concentration in shoots (31 µmol g-1). Severity of ear rot and stalk rot in maize was not significantly reduced in the amended plots. Incorporation of R. sativus 'Terranova' significantly decreased the amount of F. graminearum DNA by 58% compared with the cultivated fallow treatment, however the DNA concentration was not significantly different to fallow uncultivated. Fusarium graminearum DNA and deoxynivalenol in maize was 50% lower after incorporation of B. oleracea var. caulorapa L. compared to after fallow treatment but the difference was not significant. The brassica crops used in the present field experiment were not effective in suppressing F. graminearum, therefore further studies to optimise the current approach are recommended.

A new method to validate and compare varietal resistance and yield tolerance of sugar beet (Beta vulgaris) against the beet cyst nematode, Heterodera schachtii Schmidt.

BACKGROUND: Beet cyst nematode, Heterodera schachtii Schmidt is a global threat to sugar beet crops, and is found in every major sugar beet growing region. Annual losses due to this nematode can be severe, being estimated at €90 m in Europe alone in the 1990s. Fortunately tolerant, resistant and partially resistant varieties have since been introduced which help to limit yield loss and are now widely being deployed in infested fields. However, understanding yield performance of these varieties has been difficult, especially when variety testing programmes usually require uninfested fields. RESULTS: For the first time, and in a standardised manner, we can now assess simultaneously the resistance of different varieties to BCN and their actual yield tolerance, by comparing them to varieties grown in uninfested micro-plots alongside those which are infested. This method provides new insights on variety yield performance and nematode reproduction over an entire growing season. In addition, the investigations are also been able to detect significant physiological differences in the development and growth of the tolerant varieties' canopies and leaf chlorophyll levels. CONCLUSIONS: Our findings are of direct benefit to sugar beet growers challenged by BCN. The standardised testing provides new information on predicted variety performance. We found that these tests are justified, as not all tolerant varieties respond in the same manner to nematode infestation. Therefore, these assessments will become a vital part of variety testing for sugar beet growers, allowing for tailored deployment of variety types and more informed decision making on-farm, helping to maximise yields whilst minimising nematode damage. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Antifungal Effect of Brassica Tissues on the Mycotoxigenic Cereal Pathogen Fusarium graminearum.

Fusarium graminearum is a globally important cereal pathogen, causing head blight in wheat, resulting in yield losses and mycotoxin contamination. Currently, triazole fungicides are used to suppress Fusarium graminearum, however, the declining effectiveness of triazoles and concerns over the safety of pesticides have led to the pursuit of safe alternative crop protection strategies such as biofumigation. In the present study, species belonging to Brassicaceae (Brassica juncea, Raphanus sativus, Eruca sativa) were assessed for their biofumigation potential against F. graminearum and the glucosinolate profile of the brassicas was determined. In Petri dishes, mycelial plugs of Fusarium graminearum were exposed to frozen/defrosted leaf discs of brassicas collected at early-leaf, stem-extension, and early-bud stages. Additionally, F. graminearum inoculum was incubated in soil amended with chopped tissues of brassicas in a closed jar experiment. Glucosinolate analysis of the leaf tissue of brassicas revealed that the total glucosinolate concentration of B. juncea 'Brons' increased with advancing growth stage (24.5-51.9 µmol g-1). Brassica juncea leaf discs were effective against mycelial growth, while the sinigrin content in the leaf tissue corresponded to the level of suppression. At the stem-extension and early-bud stages, B. juncea 'Brons' showed 87-90% suppression with four leaf discs, and 100% suppression with eight leaf discs. Brassica juncea 'Caliente Rojo' leaf discs collected at the stem-extension stage showed 94% inhibition with eight discs. In the closed jar experiment, each brassica species significantly suppressed F. graminearum inoculum by 41-55%. The findings suggest that the brassica species investigated in the present study could be effective in reducing the inoculum of F. graminearum in soil prior to cereal production.

Stability of Patches of Higher Population Density within the Heterogenous Distribution of the Gray Field Slug Deroceras reticulatum in Arable Fields in the UK.

Exploitation of heterogenous distributions of Deroceras reticulatum, in arable fields by targeting molluscicide applications toward areas with higher slug densities, relies on these patches displaying sufficient spatio-temporal stability. Regular sampling of slug activity/distribution was undertaken using 1 ha rectangular grids of 100 refuge traps established in 22 commercial arable field crops. Activity varied significantly between the three years of the study, and the degree of aggregation (Taylor's Power Law) was higher in fields with higher mean trap catches. Hot spot analysis detected statistically significant spatial clusters in all fields, and in 162 of the 167 individual assessment visits. The five assessment visits in which no clusters were detected coincided with low slug activity (≤0.07 per trap). Generalized Linear Models showed significant spatial stability of patches in 11 fields, with non-significant fields also characterized by low slug activity (≤1.2 per trap). Mantel's permutation tests revealed a high degree of correlation between location of individual patches between sampling dates. It was concluded that patches of higher slug density were spatio-temporally stable, but detection using surface refuge traps (which rely on slug activity on the soil surface) was less reliable when adverse environmental conditions resulted in slugs retreating into the upper soil horizons.

Locomotor behaviour promotes stability of the patchy distribution of slugs in arable fields: Tracking the movement of individual Deroceras reticulatum.

BACKGROUND: The distribution of the grey field slug (Deroceras reticulatum Müller) in arable fields is characterised by patches containing higher slug densities dispersed within areas of lower densities. Behavioural responses that lead to the spatial/temporal stability of these patches are poorly understood, thus this study investigated behavioural mechanisms underpinning slug distribution using a new method for long-term tracking of individual slug movement in the field. RESULTS: A technique for implanting radio frequency identification (RFID) tags (each with a unique identification code) beneath the body wall of slugs was developed. Laboratory tests indicated no consistent detrimental effect on survival, feeding, egg laying or locomotor behaviour (velocity, distance travelled). Movement of individual slugs above and below the soil surface was recorded for >5 weeks (in spring and autumn) in winter wheat fields. Most (~80%) foraged within a limited area; and at the end of the observation period were located at a mean distance of 78.7 ± 33.7 cm (spring) or 101.9 ± 24.1 cm (autumn) from their release point. The maximum detected distance from the release point was 408.8 cm. The remaining slugs (~20%) moved further away and ultimately were lost. CONCLUSIONS: RFID tagging allowed continuous tracking of individual slugs, even below the soil surface. Localised movement of 80% of tracked slugs over 5 weeks offers a mechanism promoting stable slug patches in arable crops. Rapid dispersal of the remaining slugs facilitates exchange of individuals between patches. Precision targeting of pesticides at such stable slug patches may facilitate reduced usage. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Evaluating resistant brassica trap crops to manage Heterodera schachtii (Schmidt) infestations in eastern England.

BACKGROUND: The beet cyst nematode (BCN), Heterodera schachtii Schmidt, is a plant parasitic nematode that causes severe yield losses in sugar beet. Resistant brassicas (radish and mustard) have been bred to be planted after harvest of the main crop, for example a cereal, and encourage BCN juvenile hatching. The resistant plants stimulate juvenile hatching but are not suitable hosts. Juveniles are unable to complete their lifecycle and thus populations are lowered. This research aimed to investigate the effectiveness of a range of brassicas in terms of BCN control when grown in infested fields in eastern England. RESULTS: Experiments used four different radish cultivars, which differed in their resistance to BCN, and one resistant mustard variety. Field experiments were sown in early September in 2016 and 2017. Significant reductions in BCN populations were found only following resistant mustard and the radish with the greatest resistance level. CONCLUSIONS: Further research is needed to understand how best to utilize brassicas and whether they are economically viable when alternative management options for BCN are available. Time of planting may be crucial to fully achieve their BCN-reducing potential. © 2018 Society of Chemical Industry.

Beneficial effects of natural eggshell membrane versus placebo in exercise-induced joint pain, stiffness, and cartilage turnover in healthy, postmenopausal women.

PURPOSE: Despite its many health benefits, moderate exercise can induce joint discomfort when done infrequently or too intensely even in individuals with healthy joints. This study was designed to evaluate whether NEM® (natural eggshell membrane) would reduce exercise-induced cartilage turnover or alleviate joint pain or stiffness, either directly following exercise or 12 hours post exercise, versus placebo. PATIENTS AND METHODS: Sixty healthy, postmenopausal women were randomly assigned to receive either oral NEM 500 mg (n=30) or placebo (n=30) once daily for two consecutive weeks while performing an exercise regimen (50-100 steps per leg) on alternating days. The primary endpoint was any statistically significant reduction in exercise-induced cartilage turnover via the biomarker C-terminal cross-linked telopeptide of type-II collagen (CTX-II) versus placebo, evaluated at 1 and 2 weeks of treatment. Secondary endpoints were any reductions in either exercise-induced joint pain or stiffness versus placebo, evaluated daily via participant questionnaire. The clinical assessment was performed on the per protocol population. RESULTS: NEM produced a significant absolute treatment effect (TEabs) versus placebo for CTX-II after both 1 week (TEabs -17.2%, P=0.002) and 2 weeks of exercise (TEabs -9.9%, P=0.042). Immediate pain was not significantly different; however, rapid treatment responses were observed for immediate stiffness (Day 7) and recovery pain (Day 8) and recovery stiffness (Day 4). No serious adverse events occurred and the treatment was reported to be well tolerated by study participants. CONCLUSION: NEM rapidly improved recovery from exercise-induced joint pain (Day 8) and stiffness (Day 4) and reduced discomfort immediately following exercise (stiffness, Day 7). Moreover, a substantial chondroprotective effect was demonstrated via a decrease in the cartilage degradation biomarker CTX-II. Clinical Trial Registration number: NCT02751944.

Field evaluation of the nematicide fluensulfone for control of the potato cyst nematode Globodera pallida.

BACKGROUND: Three field experiments evaluated the performance of the nematicide fluensulfone against the potato cyst nematode Globodera pallida in Shropshire, England. RESULTS: Experiments 1 and 2 showed reduced root infection and lowered multiplication of G. pallida following fluensulfone (Nimitz 15G(®) ) soil treatments at five rates (1.95, 3.00, 4.05 (full rate), 5.05 and 6.00 kg AI ha(-1) ) and Nimitz 480EC(®) at the full rate. Experiment 3 demonstrated a positive interaction between the full rate of Nimitz 15G and the potato variety Santé in the reduction of G. pallida. The fluensulfone treatments at the full rate had more consistent effects than the lower rates, and there were no greater effects for the treatments higher than this full rate. Generally, fluensulfone was less efficacious than oxamyl or fosthiazate, which suggests that the treatment may not be reliably integrated within shorter potato rotations. CONCLUSION: The data suggest that fluensulfone soil application could make a useful addition to the few available nematicide treatments for the control of G. pallida rather than be a substitute for these treatments.© 2016 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effectiveness of NEM® brand eggshell membrane in the treatment of suboptimal joint function in dogs: a multicenter, randomized, double-blind, placebo-controlled study.

INTRODUCTION: Sub-optimal joint function is extremely prevalent in dogs. Therefore, a 6-week, prospective, randomized, double-blind, placebo-controlled study was conducted at eight different veterinary clinics to evaluate the efficacy, safety, and tolerability of NEM® brand eggshell membrane (EM), a novel dietary supplement shown in other species to help maintain healthy joints and connective tissues. SUBJECTS AND METHODS: Fifty-one dogs received oral EM ~13.5 mg/kg (6 mg/lb) or placebo (excipients) once daily for 6 weeks. The primary outcome measure of this study was to evaluate the change in mean joint function following 1 week and 6 weeks of supplementation as determined via the Canine Brief Pain Inventory (CBPI) questionnaire (Q#5-10) in the treatment group versus the placebo group. Secondary outcome measures were for changes in mean CBPI pain and CBPI quality of life, and mean joint pain, mobility and lameness via Veterinary Canine Scoring Assessments (VCSA). A final secondary outcome measure was for a change in serum levels of the cartilage degradation biomarker, c-terminal cross-linked telopeptide of type-II collagen (CTX-II). RESULTS: Supplementation with EM produced a significant treatment response versus placebo at 1 week (20.5% improvement, P=0.028), but fell shy of significance at 6 weeks post-treatment (22.5% improvement) for the primary outcome measure (CBPI Function), despite a sizeable treatment effect. Similarly, there was also a significant treatment response versus placebo at 1 week for CBPI Pain (19.4% improvement, P=0.010), but fell just shy of significance at 6 weeks (22.5% improvement), again despite a sizeable treatment effect. Results were not significant versus placebo at 1 week for CBPI quality of life (14.0% improvement), but produced a significant treatment response by the end of the 6-week study (26.8% improvement, P=0.033). Additionally, EM produced a significant treatment response versus placebo at 6 weeks for VCSA pain (23.6% improvement, P=0.012), but fell shy of significance for VCSA mobility and VCSA lameness (walking & trotting). Serum CTX-II levels in EM-supplemented dogs was significantly improved versus placebo at 6 weeks (47.9% improvement, P=0.018). There were no serious adverse events reported during the study and subject dog owners reported that EM was well tolerated by their pets. CONCLUSION: Supplementation with EM, ~13.5 mg/kg (6 mg/lb) taken once daily, significantly reduced joint pain and improved joint function rapidly (CBPI 1 week) and demonstrated a lasting improvement in joint pain (VCSA 6 weeks) leading to an improved quality of life (CBPI 6 weeks). Moreover, a profound chondroprotective effect was demonstrated following 6 weeks of supplementation with EM (CTX-II).

Biofumigation with Brassica juncea, Raphanus sativus and Eruca sativa for the management of field populations of the potato cyst nematode Globodera pallida.

BACKGROUND: The viability of potato cyst nematode (PCN) populations (Globodera pallida) was evaluated in three field experiments using Brassica juncea, Raphanus sativus and Eruca sativa amendments. These species were summer cultivated and autumn incorporated in experiment 1; in experiment 2, overwintered brassicaceous cover crops were spring incorporated. Experiment 3 involved determination of effects of metconazole application on biomass/glucosinolate production by B. juncea and R. sativus and on PCN pre- and post-incorporation. Glucosinolate contents were determined before incorporation. Following cover crop incorporation, field plots were planted with susceptible potatoes to evaluate the biofumigation effects on PCN reproduction. RESULTS: In experiment 1, PCN population post-potato harvest was reduced (P = 0.03) in B. juncea-treated plots, while R. sativus prevented further multiplication, but in experiment 2 there were no significant effects on PCN reproduction. In experiment 3, B. juncea or R. sativus either untreated or treated with metconazole reduced PCN populations. Glucosinolate concentrations varied significantly between different plant regions and cultivation seasons. Metconazole application increased the sinigrin concentration in B. juncea tissues. Glucosinolate concentrations correlated positively with PCN mortality for summer-cultivated brassicaceous plants. CONCLUSION: The results demonstrated that B. juncea and R. sativus green manures can play an important role in PCN management, particularly if included in an integrated pest management scheme.

Molecular studies to identify the Fusarium species responsible for HT-2 and T-2 mycotoxins in UK oats.

High levels of Fusarium mycotoxins HT-2 and T-2 have been detected in UK oats since surveys started in 2002. Fusarium langsethiae and the closely related species F. sporotrichioides have previously been associated with the contamination of cereals with type A trichothecenes HT-2 and T-2 in Nordic countries. Preliminary microbiological analysis of UK oat samples with high concentrations of HT-2 and T-2 detected and isolated F. langsethiae and F. poae but not the other type A trichothecene producing species F. sporotrichioides, F. sibiricum and F. armeniacum. Two hundred and forty oat flour samples with a known mycotoxin profile were selected from a previous four year study (2002-2005) to cover the full concentration range from below the limit of quantification (<20 µg/kg) to 9,990 µg/kg HT-2+T-2 combined. All samples were analysed for the DNA of F. langsethiae, F. poae and F. sporotrichioides based on previously published PCR assays. F. langsethiae was detectable in nearly all samples; F. poae was detected in 90% of samples whereas F. sporotrichioides was not detected in any sample. A real-time PCR assay was developed to quantify F. langsethiae DNA in plant material. The assay could quantify as low as 10(-4)ngF. langsethiae DNA/µl. Based on this assay and a previously published assay for F. poae, both species were quantified in the oat flour samples with known HT-2+T-2 content. Results showed a good regression (P<0.001, r(2)=0.60) between F. langsethiae DNA and HT-2+T-2 concentration. F. poae DNA concentration was not correlated to HT2+T2 concentration (P=0.448) but was weakly correlated to nivalenol concentration (P<0.001, r(2)=0.09). Multiple regression with F. langsethiae and F. poae DNA as explanatory variates identified that both F. langsethiae and F. poae DNA were highly significant (P<0.001) but F. poae DNA only accounted for an additional 4% of the variance and the estimate was negative, indicating that higher concentrations of F. poae DNA were correlated with slightly lower concentrations of HT2+T2 detected. A stronger regression (P<0.001, r(2)=0.77) between F. langsethiae DNA and HT-2+T-2 was obtained after extraction and quantification of DNA and mycotoxins from individual oat grains. The results from this study provide strong evidence that F. langsethiae is the primary, if not sole, fungus responsible for high HT-2 and T-2 in UK oats.