Victor Limay-Rios

Neonicotinoid Insecticide Residues in Surface Water and Soil Associated with Commercial Maize (Corn) Fields in Southwestern Ontario

Neonicotinoid insecticides have come under scrutiny for their potential unintended effects on non-target organisms, particularly pollinators in agro-ecosystems. As part of a larger study of neonicotinoid residues associated with maize (corn) production, 76 water samples within or around the perimeter of 18 commercial maize fields and neighbouring apiaries were collected in 5 maize-producing counties of southwestern Ontario. Residues of clothianidin (mean = 2.28, max. = 43.60 ng/mL) and thiamethoxam (mean = 1.12, max. = 16.50 ng/mL) were detected in 100 and 98.7% of the water samples tested, respectively. The concentration of total neonicotinoid residues in water within maize fields increased six-fold during the first five weeks after planting, and returned to pre-plant levels seven weeks after planting. However, concentrations in water sampled from outside the fields were similar throughout the sampling period. Soil samples from the top 5 cm of the soil profile were also collected in these fields before and immediately following planting. The mean total neonicotinoid residue was 4.02 (range 0.07 to 20.30) ng/g, for samples taken before planting, and 9.94 (range 0.53 to 38.98) ng/g, for those taken immediately after planting. Two soil samples collected from within an conservation area contained detectable (0.03 and 0.11 ng/g) concentrations of clothianidin. Of three drifted snow samples taken, the drift stratum containing the most wind-scoured soil had 0.16 and 0.20 ng/mL mainly clothianidin in the melted snow. The concentration was at the limit of detection (0.02 ng/mL) taken across the entire vertical profile. With the exception of one sample, water samples tested had concentrations below those reported to have acute, chronic or sublethal effects to honey bees. Our results suggest that neonicotinoids may move off-target by wind erosion of contaminated soil. These results are informative to risk assessment models for other non-target species in maize agro-ecosytems.

Neonicotinoid insecticide residues in soil dust and associated parent soil in fields with a history of seed treatment use on crops in Southwestern Ontario

Using neonicotinoid insecticides as seed treatments is a common practice in field crop production. Exposure of nontarget organisms to neonicotinoids present in various environmental matrices is debated. In the present study, concentrations of neonicotinoid residues were measured in the top 5 cm of soil and overlying soil surface dust before planting in 25 commercial fields with a history of neonicotinoid seed treatment use in southwestern Ontario in 2013 and 2014 using liquid chromatography-electrospray ionization tandem mass spectrometry. The mean total concentrations were 3.05 ng/g and 47.84 ng/g in 2013 and 5.59 ng/g and 71.17 ng/g in 2014 for parent soil and soil surface dust, respectively. When surface and parent soil residues were compared the mean concentration in surface dust was 15.6-fold and 12.7-fold higher than that in parent soil in 2013 and 2014, respectively. Pooled over years, the surface dust to parent soil ratio was 13.7, with mean concentrations of 4.36 ng/g and 59.86 ng/g for parent soil and surface dust, respectively. The present studys results will contribute important knowledge about the role these residues may play in the overall risk assessment currently under way for the source, transport, and impact of neonicotinoid insecticide residues in a maize ecosystem.

Field-scale examination of neonicotinoid insecticide persistence in soil as a result of seed treatment use in commercial maize (corn) fields in southwestern Ontario

Neonicotinoid insecticides, especially as seed treatments, have raised concerns about environmental loading and impacts on pollinators, biodiversity, and ecosystems. The authors measured concentrations of neonicotinoid residues in the top 5 cm of soil before planting of maize (corn) in 18 commercial fields with a history of neonicotinoid seed treatment use in southwestern Ontario in 2013 and 2014 using liquid chromatography-tandem mass spectrometry with electrospray ionization. A simple calculator based on first-order kinetics, incorporating crop rotation, planting date, and seed treatment history from the subject fields, was used to estimate dissipation rate from the seed zone. The estimated half-life (the time taken for 50% of the insecticide to have dissipated by all mechanisms) based on 8 yr of crop history was 0.64 (range, 0.25-1.59) yr and 0.57 (range, 0.24-2.12) yr for 2013 and 2014, respectively. In fields where neonicotinoid residues were measured in both years, the estimated mean half-life between 2013 and 2014 was 0.4 (range, 0.27-0.6) yr. If clothianidin and thiamethoxam were used annually as a seed treatment in a typical crop rotation of maize, soybean, and winter wheat over several years, residues would plateau rather than continue to accumulate. Residues of neonicotinoid insecticides after 3 yr to 4 yr of repeated annual use tend to plateau to a mean concentration of less than 6 ng/g in agricultural soils in southwestern Ontario.

Bacterial endophytes from wild maize suppress Fusarium graminearum in modern maize and inhibit mycotoxin accumulation

Wild maize (teosinte) has been reported to be less susceptible to pests than their modern maize (corn) relatives. Endophytes, defined as microbes that inhabit plants without causing disease, are known for their ability to antagonize plant pests and pathogens. We hypothesized that the wild relatives of modern maize may host endophytes that combat pathogens. Fusarium graminearum is the fungus that causes Gibberella Ear Rot (GER) in modern maize and produces the mycotoxin, deoxynivalenol (DON). In this study, 215 bacterial endophytes, previously isolated from diverse maize genotypes including wild teosintes, traditional landraces and modern varieties, were tested for their ability to antagonize F. graminearum in vitro. Candidate endophytes were then tested for their ability to suppress GER in modern maize in independent greenhouse trials. The results revealed that three candidate endophytes derived from wild teosintes were most potent in suppressing F. graminearum in vitro and GER in a modern maize hybrid. These wild teosinte endophytes could suppress a broad spectrum of fungal pathogens of modern crops in vitro. The teosinte endophytes also suppressed DON mycotoxin during storage to below acceptable safety threshold levels. A fourth, less robust anti-fungal strain was isolated from a modern maize hybrid. Three of the anti-fungal endophytes were predicted to be Paenibacillus polymyxa, along with one strain of Citrobacter. Microscopy studies suggested a fungicidal mode of action by all four strains. Molecular and biochemical studies showed that the P. polymyxa strains produced the previously characterized anti-Fusarium compound, fusaricidin. Our results suggest that the wild relatives of modern crops may serve as a valuable reservoir for endophytes in the ongoing fight against serious threats to modern agriculture. We discuss the possible impact of crop evolution and domestication on endophytes in the context of plant defense.

Root-hair endophyte stacking in finger millet creates a physicochemical barrier to trap the fungal pathogen Fusarium graminearum.

The ancient African crop, finger millet, has broad resistance to pathogens including the toxigenic fungus Fusarium graminearum. Here, we report the discovery of a novel plant defence mechanism resulting from an unusual symbiosis between finger millet and a root-inhabiting bacterial endophyte, M6 (Enterobacter sp.). Seed-coated M6 swarms towards root-invading Fusarium and is associated with the growth of root hairs, which then bend parallel to the root axis, subsequently forming biofilm-mediated microcolonies, resulting in a remarkable, multilayer root-hair endophyte stack (RHESt). The RHESt results in a physical barrier that prevents entry and/or traps F. graminearum, which is then killed. M6 thus creates its own specialized killing microhabitat. Tn5-mutagenesis shows that M6 killing requires c-di-GMP-dependent signalling, diverse fungicides and resistance to a Fusarium-derived antibiotic. Further molecular evidence suggests long-term host–endophyte–pathogen co-evolution. The end result of this remarkable symbiosis is reduced deoxynivalenol mycotoxin, potentially benefiting millions of subsistence farmers and livestock. Further results suggest that the anti-Fusarium activity of M6 may be transferable to maize and wheat. RHESt demonstrates the value of exploring ancient, orphan crop microbiomes.

Quantifying Neonicotinoid Insecticide Residues Escaping during Maize Planting with Vacuum Planters.

Neonicotinoid residues escaping in vacuum-planter exhaust during maize planting were measured in 25 fields in southwestern Ontario in 2013-2014 using filter bags to collect planter exhaust dust and horizontal and vertical sticky traps to collect planter operation-generated dust. Atrazine residues were used to differentiate between neonicotinoid residues originating from seed or from disturbed soil. Recovery rates of seed-applied neonicotinoids in exhaust were 0.014 and 0.365% in 2013 and 2014, respectively, calculated on the basis of neonicotinoid concentrations in preplant soil and seed application rates. Neonicotinoid exhaust emission rates were 0.0036 and 0.1104 g/ha for 2013 and 2014, respectively, with 99.9472 and 99.7820% originating from treated seed in 2013 and 2014, respectively, calculated on the basis of the atrazine marker. Rates of recovery of seed-applied neonicotinoid residues by exhaust filter bags were 0.015 and 0.437% for 2013 and 2014, respectively. Neonicotinoid residues captured on horizontal and vertical traps were 1.10 ng/cm2 (0.1104 g/ha) and 1.45 ng/cm2 (0.0029 g/ha), respectively, with 92.31 and 93.03% originating from treated seed, respectively, representing 0.3896% of the original active ingredient applied to the seed planted. Exposure to pollinators can be best reduced by strategies to keep active ingredient on the seed, below the soil surface, and in the field where applied.

Occurrence of Penicillium verrucosum, ochratoxin A, ochratoxin B and citrinin in on-farm stored winter wheat from the Canadian Great Lakes Region

The occurrence of P. verrucosum and ochratoxin A (OTA) were surveyed for 3 and 4 years, respectively. A total of 250 samples was collected from an average of 30 farms during the 2011, 2012, 2013 and 2014 winter seasons. Most storage bins surveyed were typically 11 m high round bins made of corrugated, galvanized steel, with flat-bottoms and conical roofs. Samples of clumped grain contained the most P. verrucosum (p<0.05, n = 10) followed by samples taken from the first load (n = 24, mean = 147±87 CFU/g) and last load (n = 17, mean = 101±77 CFU/g). Five grain samples (2.2%) tested positive for OTA, citrinin and OTB at concentrations of 14.7±7.9, 4.9±1.9 and 1.2±0.7 ng/g, with only three samples exceeding 5 ng/g. Grain samples positive for OTA were related to moisture resulting from either condensation or migrating moist warm air in the bin or areas where precipitation including snow entered the bin. Bins containing grain and clumps contaminated with OTA were studied in detail. A number of statistically-significant risk factors for OTA contamination were identified. These included 1) grain clumps accumulated around or directly under manhole openings, 2) debris and residue of old grain or grain clumps collected from the bin walls or left on storage floor and augers and 3) grain clumps accumulated around side doors. Even when grain enters storage below the 14.5% threshold of moisture, condensation and moisture migration occurs in hotspots in modern corrugated steel storage bins. Hot spots of OTA contamination were most often in areas affected by moisture migration due to inadequate aeration and exposure to moisture from precipitation or condensation. Further, we found that the nature of the condensation affects the nature and distribution of small and isolated areas with high incidence of toxin contamination and/or P. verrucosum prevalence in the grain bins examined.

The role of field dust in pesticide drift when pesticide‐treated maize seeds are planted with vacuum‐type planters

BACKGROUND Neonicotinoid-contaminated dust escaping pneumatic seeders causes exposure to non-target organisms such as pollinators. Two sources of dust have been reported: abrasion by talc which is added as seed lubricant during planting, and seed-to-seed abrasion occurring during seed handling, distribution and planting. We report a third important source that warrants remediation. Here, soil dust stirred up by planters was found to enter the vacuum air intake near seed metering devices. RESULTS The mean quantity of dust collected from the exhaust of a commercial pneumatic planter over a number of field sites and situations was 46 g ha-1 , ranging from 5.8 to 184.2 g ha-1 . While the clothianidin concentration in exhaust dust declined with increasing quantity of dust, total clothianidin recovered increased linearly within the study parameters. Up to 2.4 g ha-1 of clothianidin was recovered from planter exhaust, representing approximately 12.6% of the active ingredient applied to seed. A similar pattern occurred in the laboratory on a single standing planter unit using diatomaceous earth as surrogate field dust. CONCLUSION Field dust in pneumatic metering systems contributes significantly to clothianidin contamination in planter exhaust by seed abrasion. Adding diatomaceous earth as surrogate field dust to the Heubach seed dust protocol accounted for field dust abrasion and distinguished anti-abrasive properties of seed treatments.

Fusarium graminearum Mycotoxins in Maize Associated With Striacosta albicosta (Lepidoptera: Noctuidae) Injury

Abstract Western bean cutworm, Striacosta albicosta (Smith; Lepidoptera: Noctuidae) has become a key pest of maize, Zea mays (L.), in Ontario, Canada which is challenging to control due to its lack of susceptibility to most Bt-maize events. Injury by S. albicosta may exacerbate Fusarium graminearum (Schwabe; Hypocreales: Nectriaceae) infection through provision of entry points on the ear. The objectives of this study were to: investigate the relationship between injury by S. albicosta and deoxynivalenol (DON) accumulation; evaluate non-Bt and Bt-maize hybrids, with and without insecticide and fungicide application; and determine optimal insecticide-fungicide application timing for reducing S. albicosta injury and DON accumulation. The incidence of injury by S. albicosta and ear rot severity were found to increase DON concentrations under favorable environmental conditions for F. graminearum infection. Incidence of S. albicosta injury was more important than severity of injury for DON accumulation which may be due to larval consumption of infected kernels. The Vip3A × Cry1Ab event provided superior protection from the incidence and severity of S. albicosta injury compared to non-Bt or Cry1F hybrids. Insecticide application to a Vip3A × Cry1Ab hybrid did not reduce injury further; however, lower severity of injury was observed for non-Bt and Cry1F hybrids when pyrethroids or diamides were applied at early VT or R1 stages. DON concentrations were reduced with application of prothioconazole fungicide tank-mixed with insecticide at late VT (before silk browning) or when insecticide was applied at early VT followed by prothioconazole at R1. The application of an insecticide/fungicide tank-mix is the most efficient approach for maize hybrids lacking high-dose insecticidal proteins against S. albicosta and F. graminearum tolerance. Results demonstrate that reducing the risk of DON accumulation requires a strategic approach to manage complex associations among S. albicosta, F. graminearum and the environment.

Effect of Prothioconazole Application Timing on Fusarium Mycotoxin Content in Maize Grain

In 2010 and 2011, studies to determine the optimal timing of prothioconazole application (200 g a.i./ha) for reducing Fusarium mycotoxin accumulation in grain were conducted in controlled replicated experiments under small-plot mist-irrigated experiments and in field-scale experiments using two hybrids susceptible to F. gramineaerum infection. A significant decrease in total deoxynivalenol (DON) [DON + 15-acetyl-DON + DON 3-glucoside + 3-acetyl-DON] and zearalenone concentrations was observed when fungicide was sprayed at VT (tasseling) and R1 (silking; P < 0.01) followed by applications at V18 (18th leaf) and R2 (blister; P < 0.05) stages, corresponding to silk completely emerged and fully elongated and to silk emergence and browning, respectively. No reduction in Fusarium graminearum toxins was found after silk senescence (R3 or milk) stage. Moniliformin, fumonisins, beauvericin, enniatins, HT-2 and T-2 toxins were also found in small quantities, and no reduction was observed after treatment ( P > 0.05). Mean reduction (±s.d.) of 59 ± 20% and 57 ± 38% of total DON and zearalenone was observed at full silk elongation, respectively.