Synthetic approaches to the 2015-2018 new agrochemicals.

In this review, the synthesis of 33 agrochemicals that received an international standardization organization (ISO) name between January 2015 and December 2018 is described. The aim is to showcase the broad range and scope of reactions, reagents and intermediates used to discover and produce the latest active ingredients addressing the crop protection industry's needs.

Establishing health-based biological exposure limits for pesticides: A proof of principle study using mancozeb.

Pesticides represent an economical, labor-saving, and efficient tool for pest management, but their intrinsic toxic properties may endanger workers and the general population. Risk assessment is necessary, and biological monitoring represents a potentially valuable tool. Several international agencies propose biological exposure indices (BEI), especially for substances which are commonly absorbed through the skin. Biological monitoring for pesticide exposure and risk assessment seems a natural choice, but biological exposure limits (BEL) for pesticides are lacking. This study aims at establishing equivalent biological exposure limits (EBEL) for pesticides using real-life field data and the Acceptable Operator Exposure Level (AOEL) of mancozeb as the reference. This study included a group of 16 vineyard pesticide applicators from Northern Italy, a subgroup of a more extensive study of 28 applicators. Their exposure was estimated using "patch" and "hand-wash" methodologies, together with biological monitoring of free ethylene-bis-thiourea (ETU) excretion in 24-h pre- and post-exposure urine samples. Modeling was done using univariate linear regression with ETU excretion as the dependent variable and the estimated absorbed dose as the independent variable. The median skin deposition of mancozeb in our study population was 125 µg, leading to a median absorbed dose of 0.9 µg/kg. The median post-exposure ETU excretion was 3.7 µg. The modeled EBEL for mancozeb was 148 µg of free ETU or 697 µg of total ETU, accounting for around 75% of the maximum theoretical excretion based on a mass balance model. Although preliminary and based on a small population of low-exposed workers, our results demonstrate a procedure to develop strongly needed biological exposure limits for pesticides.

Modeling Yield Losses and Fungicide Profitability for Managing Fusarium Head Blight in Brazilian Spring Wheat.

Fusarium head blight (FHB) and wheat yield data were gathered from fungicide trials to explore their relationship. Thirty-seven studies over 9 years and 11 locations met the criteria for inclusion in the analysis: FHB index in the untreated check ≥ 5% and the range of index in a trial ≥ 4 percentage points. These studies were grouped into two baseline yields, low (Yl ≤ 3,631 kg ha-1) or high (Yh > 3,631 kg ha-1), defined based on the median of maximum yields across trials. Attainable (disease-free) yields and FHB index were predicted using a wheat crop and a disease model, respectively, in 280 simulated trials (10 planting dates in a 28-year period, 1980 to 2007) for the Passo Fundo location. The damage coefficient was then used to calculate FHB-induced yield loss (penalizing attainable yield) for each experiment. Losses were compared between periods defined as before and after FHB resurge during the early 1990s. Disease reduction from the use of one or two sprays of a triazole fungicide (tebuconazole) was also simulated, based on previous meta-analytic estimates, and the response in yield was used in a profitability analysis. Population-average intercepts but not the slopes differed significantly between Yl (2,883.6 kg ha-1) and Yh (4,419.5 kg ha-1) baseline yields and the damage coefficients were 1.60%-1 and 1.05%-1, respectively. The magnitudes and trends of simulated yield losses were in general agreement with literature reports. The risk of not offsetting the costs of one or two fungicide sprays was generally higher (>0.75) prior to FHB resurgence but fungicide profitability tended to increase in recent years, depending on the year. Our simulations allowed us to reproduce trends in historical losses, and may be further adjusted to test the effect and profitability of different control measures (host resistance, other fungicides, etc.) on quality parameters such as test weight and mycotoxin contamination, should the information become available.

Quantifying Control Efficacy of Fungicides Commonly Applied for Potato Early Blight Management.

Early blight is an economically important foliar disease of potato in the United States. Because of the lack of resistant potato cultivars, fungicides are applied extensively to obtain adequate control. To manage early blight, standard protectant fungicides and single-site mode-of-action "specialty" fungicides are applied either alone or incorporated into a fungicide rotation program. Control efficacy at two crop growth stages (tuber initiation/early bulking and late bulking/tuber maturation) and the overall tuber yield response to standard and specialty fungicides were assessed using network metaanalytic models. Control efficacy of fungicides ranged from moderate to very high (>30 to 75%) compared with the nontreated control. For both potato growth stages, specialty fungicides performed better than standard protectant fungicides. Furthermore, control efficacy of both fungicides was higher (3 to 9%) at late bulking and tuber maturation when compared with early bulking crop growth stage. Specialty fungicide programs increased overall tuber yields by 4 and 9% over standard fungicides and nontreated control, respectively. Based on the results, more precise fungicide use recommendations and fungicide programs can be developed for early blight management.

A Quantitative Synthesis of the Efficacy and Profitability of Conventional and Biological Fungicides for Botrytis Fruit Rot Management on Strawberry in Florida.

Botrytis fruit rot (BFR) is a major disease that affects strawberry production in Florida and worldwide. BFR management relies on frequent fungicide applications. A meta-analysis was conducted on the outcomes from nine field trials to evaluate the efficacy and profitability of conventional and biological fungicides compared with a nontreated control (NTC). All trials were conducted in Florida between the 2005/06 and 2016/17 growing seasons. Fungicide treatments were applied weekly, and plots were harvested twice a week for yield and BFR incidence quantification. Treatments were grouped into four categories: NTC, multisite only (Thiram), Standard (captan alternated with fludioxonil + cyprodinil), and Bacillus. Following primary analyses, a random effects network meta-analytical model was fitted to estimate the mean yield and BFR incidence responses for each treatment group and to compare means between pairs of groups. The Thiram and the Standard treatment groups increased yield by 378.8 and 502.2 kg/ha/week, respectively, compared with the NTC. The yield difference between Bacillus and NTC was not statistically significant. Besides increasing yield, Thiram and Standard also reduced BFR incidence by approximately 10% compared with the NTC. The mean yield responses and among-study variability from the meta-analysis were used to estimate the probability of a given yield response in a new future trial. The Standard and Thiram treatment groups showed higher estimated probabilities of increasing yield and resulting in a profitable return on application investments than the Bacillus group of treatments. The results from this study provide growers with information that will aid their decision-making process regarding BFR management.

Performance and Profitability of Fungicides for Managing Soybean White Mold: A 10-Year Summary of Cooperative Trials.

White mold, caused by Sclerotinia sclerotiorum, is a yield-limiting disease of soybean in Brazil. Uniform fungicide trials have been conducted annually since 2009. Data from 74 cooperative field trials conducted over a 10-year period were assembled. We selected five fungicides applied two times around flowering: dimoxystrobin plus boscalid (DIMO+BOSC), carbendazim plus procymidone (CARB+PROC), fluazinam (FLUZ), fluopyram (FLUO), and procymidone (PROC). For comparison, thiophanate-methyl (TMET) applied four times was also included as a low-cost treatment. Network models were fitted to the log of white mold incidence (percentages) and log of sclerotia mass data (grams/hectare) and to the nontransformed yield data (kilograms/hectare) for each treatment, including the untreated check. Back-transformation of the meta-analytic estimates indicated that the lowest and highest mean (95% confidence interval [CI]) percent reductions in incidence and sclerotia mass were 54.2 (49.3 to 58.7) and 51.6% (43.7 to 58.3) for TMET and 83.8 (79.1 to 87.5) and 87% (81.9 to 91.6) for CARB+PROC, respectively. The overall mean (95% CI) yield responses ranged from 323 kg/ha (247.4 to 400.3) for TMET to 626 kg/ha (521.7 to 731.7) for DIMO+BOSC, but the variance was significantly reduced by a binary variable (30% threshold) describing disease incidence in the untreated check. On average, an increment of 352 kg/ha was estimated for trials where the incidence was >30% compared with the low-disease scenario. Hence, the probability of breaking even on fungicide costs for the high-disease scenario was >65% for the more effective, but more expensive fungicide (FLUZ) than TMET. For the low-disease scenario, profitability was less likely and depended more on variations in fungicide cost and soybean price.

Determination and validation of an aquatic Maximum Acceptable Concentration-Environmental Quality Standard (MAC-EQS) value for the agricultural fungicide azoxystrobin.

The main goal of the present study was to determine and validate an aquatic Maximum Acceptable Concentration-Environmental Quality Standard (MAC-EQS) value for the agricultural fungicide azoxystrobin (AZX). Assessment factors were applied to short-term toxicity data using the lowest EC50 and after the Species Sensitivity Distribution (SSD) method. Both ways of EQS generation were applied to a freshwater toxicity dataset for AZX based on available data, and to marine toxicity datasets for AZX and Ortiva® (a commercial formulation of AZX) obtained by the present study. A high interspecific variability in AZX sensitivity was observed in all datasets, being the copepoda Eudiaptomus graciloides (LC50,48h = 38 µg L-1) and the gastropod Gibbula umbilicalis (LC50,96h = 13 µg L-1) the most sensitive freshwater and marine species, respectively. MAC-EQS values derived using the lowest EC50 (≤0.38 µg L-1) were more protective than those derived using the SSD method (≤3.2 µg L-1). After comparing the MAC-EQS values estimated in the present study to the smallest AA-EQS available, which protect against the occurrence of prolonged exposure of AZX, the MAC-EQS values derived using the lowest EC50 were considered overprotective and a MAC-EQS of 1.8 µg L-1 was validated and recommended for AZX for the water column. This value was derived from marine toxicity data, which highlights the importance of testing marine organisms. Moreover, Ortiva affects the most sensitive marine species to a greater extent than AZX, and marine species are more sensitive than freshwater species to AZX. A risk characterization ratio higher than one allowed to conclude that AZX might pose a high risk to the aquatic environment. Also, in a wider conclusion, before new pesticides are approved, we suggest to improve the Tier 1 prospective Ecological Risk Assessment by increasing the number of short-term data, and apply the SSD approach, in order to ensure the safety of aquatic organisms.

Comparing effect levels of regulatory studies with endpoints derived in targeted anti-androgenic studies: example prochloraz.

Prochloraz is an imidazole fungicide, and its regulatory toxicological data package has been primarily generated in the 1990s. More recently, studies have been published demonstrating an interaction with hormone receptors/steroidogenesis and effects with an endocrine mode of action. In the present study, prochloraz has been investigated in a comprehensive in vivo study including relevant elements of current regulatory reproduction toxicity studies and additional mechanistic parameters. Prochloraz was administered per gavage in oil from GD 6 to PND 83 to pregnant and lactating Wistar rats and their respective offspring, at doses of 0.01 mg/kg bw/day (acceptable daily intake of prochloraz), 5 mg/kg bw/day [expected no-observed-effect-level (NOEL)] and 30 mg/kg bw/day. At 30 mg/kg bw/day maternal and offspring effects (decreased viability, lower number of live offspring) were seen including a delayed entry into male puberty (+1 day) accompanied by lower male offspring body weights, increased anogenital distance/index in females and transiently retained nipples in males at PND 12 (not seen at PND 20). The only finding at the "expected NOEL" was increased incidences of transiently retained nipples in males which are not considered adverse. No effects were seen in the low-dose group. There was no evidence for a non-monotonic dose-response curve or effects at low levels.

Enhanced Overwintering Survival of Stevia by QoI Fungicides Used for Management of Sclerotium rolfsii.

Stevia (Stevia rebaudiana) is a herbaceous perennial under evaluation as a new crop in the southeastern United States. Stem rot caused by Sclerotium rolfsii is common in stevia plantings in North Carolina, with symptoms including wilting, root and stem necrosis, and plant death. Fungicide efficacy trials for management of S. rolfsii were conducted over 2 years. Fungicides evaluated included azoxystrobin, flutolanil, and tebuconazole applied at three timings. Azoxystrobin applied to transplants 1 week prior to planting had the lowest area under the disease progress curve values across all trials. Fungicide plots were also used to evaluate overwintering of stevia. End-of-season stand counts were compared with spring emergence counts to quantify overwintering survival. In spring 2015, plots treated with azoxystrobin in 2014 had greater overwintering survival (78%) than other fungicide treatments (38%) and the control (38%). Similar results were obtained at two locations in spring 2016 in plots treated with azoxystrobin or pyraclostrobin in 2015. Successful overwintering of stevia directly impacts the profitability of second- and third-year harvests and enhances the likelihood of long-term establishment of stevia as a viable crop. Future studies will be directed at elucidating the mechanism of the enhanced overwintering survival of plants treated with quinone outside inhibitor fungicides.

A predictive data-driven framework for endocrine prioritization: a triazole fungicide case study.

The US Environmental Protection Agency Endocrine Disruptor Screening Program (EDSP) is a tiered screening approach to determine the potential for a chemical to interact with estrogen, androgen, or thyroid hormone systems and/or perturb steroidogenesis. Use of high-throughput screening (HTS) to predict hazard and exposure is shifting the EDSP approach to (1) prioritization of chemicals for further screening; and (2) targeted use of EDSP Tier 1 assays to inform specific data needs. In this work, toxicology data for three triazole fungicides (triadimefon, propiconazole, and myclobutanil) were evaluated, including HTS results, EDSP Tier 1 screening (and other scientifically relevant information), and EPA guideline mammalian toxicology study data. The endocrine-related bioactivity predictions from HTS and information that satisfied the EDSP Tier 1 requirements were qualitatively concordant. Current limitations in the available HTS battery for thyroid and steroidogenesis pathways were mitigated by inclusion of guideline toxicology studies in this analysis. Similar margins (3-5 orders of magnitude) were observed between HTS-predicted human bioactivity and exposure values and between in vivo mammalian bioactivity and EPA chronic human exposure estimates for these products' registered uses. Combined HTS hazard and human exposure predictions suggest low priority for higher-tiered endocrine testing of these triazoles. Comparison with the mammalian toxicology database indicated that this HTS-based prioritization would have been protective for any potential in vivo effects that form the basis of current risk assessment for these chemicals. This example demonstrates an effective, human health protective roadmap for EDSP evaluation of pesticide active ingredients via prioritization using HTS and guideline toxicology information.

Agricultural Applications for Antimicrobials. A Danger to Human Health: An Official Position Statement of the Society of Infectious Diseases Pharmacists.

The use of antibiotics in agriculture, particularly in food-producing animals, is pervasive and represents the overwhelming majority of antibiotic use worldwide. The link between antibiotic use in animals and antibiotic resistance in humans is unequivocal. Transmission can occur by ingesting undercooked meats harboring resistant bacteria, by direct contact of animals by animal handlers, and by various other means. Antibiotics used in aquaculture and antifungals used in horticulture are also an evolving threat to human health. Regulations aimed at decreasing the amount of antibiotics used in food production to limit the development of antibiotic resistance have recently been implemented. However, further action is needed to minimize antibiotic use in agriculture. This article describes the extent of this current problem and serves as the official position of the Society of Infectious Diseases Pharmacists on this urgent threat to human health.

Synthetic approaches to the 2010-2014 new agrochemicals.

In this review, the synthesis of 30 agrochemicals that received an international standardization organization (ISO) name during the last five years (January 2010 to December 2014) is described. The aim is to showcase the range and scope of chemistries used to discover or produce the latest active ingredients addressing the crop protection industry's needs.

Lethal and behavioral effects of pesticides on the insect predator Macrolophus pygmaeus.

Macrolophus pygmaeus (Hemiptera: Miridae) is a common generalist predator in Mediterranean agro-ecosystems. We evaluated the lethal effects of six insecticides and a fungicide on M. pygmaeus nymphs exposed to the pesticides through three routes of exposure: direct, residual and oral. Chlorantraniliprole and emamectin-benzoate caused less than 25% mortality to M. pygmaeus and were classified as harmless according to the International Organization for Biological Control rating scheme. In contrast, thiacloprid and metaflumizone caused 100% and 80% mortality, respectively, and were classified as harmful. Indoxacarb and spinosad resulted in close to 30% mortality to the predator, and were classified as slightly harmful, while the fungicide copper hydroxide caused 58% mortality and was rated as moderately harmful. Chlorantraniliprole and thiacloprid were selected for further sublethal testing by exposing M. pygmaeus to two routes of pesticide intake: pesticide residues and feeding on sprayed food. Thiacloprid led to an increase in resting and preening time of the predator, and a decrease in plant feeding. Chlorantraniliprole resulted in a decrease in plant feeding, but no other behaviors were affected. In addition, thiacloprid significantly reduced the predation rate of M. pygmaeus, whereas chlorantraniliprole had no significant effect on predation rate. The results of the study suggest that thiacloprid is not compatible with M. pygmaeus, while further research needs to be carried out for metaflumizone and copper hydroxide. All other products seem to be relatively compatible with M. pygmaeus, though studies on their sublethal effects will shed more light into their safety.

A quantitative solid-state Raman spectroscopic method for control of fungicides.

A new analytical procedure using solid-state Raman spectroscopy within the THz-region for the quantitative determination of mixtures of different conformations of trifloxystrobin (EE, EZ, ZE and ZZ), tebuconazole (1), and propiconazole (2) as an effective method for the fungicide product quality monitoring programmes and control has been developed and validated. The obtained quantities were controlled independently by the validated hybrid HPLC electrospray ionization (ESI) tandem mass spectrometric (MS) and matrix-assisted laser desorption/ionization (MALDI) MS methods in the condensed phase. The quantitative dependences were obtained on the twenty binary mixtures of the analytes and were further tested on the three trade fungicide products, containing mixtures of trifloxystrobin-tebuconazole and trifloxystrobin-propiconazole, as an emissive concentrate or water soluble granules of the active ingredients. The present methods provided sufficient sensitivity as reflected by the metrologic quantities, evaluating the concentration limit of detection (LOD) and quantification (LOQ), linear limit (LL), measurement accuracy and precision, true quantity value, trueness of measurement and more.

Debilitation in conidia of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae and implication with respect to viability determinations and mycopesticide quality assessments.

Germination of Beauveria bassiana (Bb) and Metarhizium anisopliae (Ma) conidia determined from a fast-rehydration (FR) protocol were compared to those obtained when dry conidia were subjected to slow rehydration (SR) by holding under high humidity conditions prior to aqueous suspension. Differences in viability estimates obtained using the FR vs. SR protocols increased markedly after conidia were exposed to various stress factors in storage (high a(w), temperature, and O(2) concentrations), with the SR protocol producing higher estimates of viability in all cases. After Bb conidia were stored under moist conditions for 21 days at 25 degrees C, the SR estimate of viability was >21% greater than the FR estimate. In jars flushed with different O(2) concentrations and stored at 50 degrees C for 34 days, proportional differences between protocols varied, depending on water activity, from 18-44% in jars flushed with 0% O(2) (100% N(2)) to as high as 63-93% when treated with 21-22% O(2). For conidia stored over a broad range of moderate to high temperatures in the absence of O(2), SR-FR differences were

Biomonitoring equivalents for hexachlorobenzene.

Recent efforts worldwide have resulted in a growing database of measured concentrations of chemicals in blood and urine samples taken from the general population. However, few tools exist to assist in the interpretation of the measured values in a health risk context. Biomonitoring equivalents (BEs) are defined as the concentration or range of concentrations of a chemical or its metabolite in a biological medium (blood, urine, or other medium) that is consistent with an existing health-based exposure guideline. This study reviews available health-based exposure guidance values for hexachlorobenzene (HCB) from Health Canada, the United States Environmental Protection Agency (US EPA), the US Agency for Toxic Substances and Disease Registry (ATSDR) and World Health Organization (WHO). HCB liver tissue concentrations in chronic rodent bioassays and information on human elimination rates and tissue distribution of HCB were extrapolated to estimate serum lipid-adjusted HCB concentrations that are consistent with the exposure guidance values for HCB. Estimated serum lipid-adjusted HCB concentrations ranging from 16 to 250 ng/g lipid were consistent with non-cancer-based exposure guidance values from various agencies. Concentrations associated with cancer risk-specific doses at target risk levels of interest were also estimated. These BE values may be used as screening tools for evaluation of population biomonitoring data for HCB in a risk assessment context and can assist in prioritization of the potential need for additional risk assessment efforts for HCB relative to other chemicals.

Toxicity of copper/dimethomorph combination for Lemna minor and depuration of the fungicides by aquatic plant.

Runoff-water of Champagne vineyards is often collected in decantation ponds where pesticides may be eliminated before the water is transferred to rivers. In order to accelerate this process, Lemna minor was tested for its phytoremediation capacity. In the ponds several pesticides coexist. Therefore, the cross-influence of copper and dimethomorph, two pesticides frequently used on grape to control fungal diseases, was studied on toxicity and their removal by L. minor. The toxicity of copper and dimethomorph alone and in combination, was assessed by growth inhibition of L. minor cultures after 96 h and 168 h. Copper had a severe impact on growth (maximum inhibition: 90% at 1000 microg/L) while dimethomorph (as pure ingredient DMM(P) or formulated as Forum DMM(F)) did not (inhibition < 45% at 1000 microg/L) after 168 h of treatment. When both chemicals were combined, a synergistic effect was detected after 96 h of exposure to copper and DMM(F). However, this interaction tended towards additivity after 168 h. Additivity was also observed when DMM(P) replaced DMM(F) in the mixture of copper and dimethomorph at 96h00 and 168h00. The removal of copper depended on DMM(F) concentration. For example, with 250 microg/L of dimethomorph as Forum, removal of copper increased from 36 to 41%. Depuration of DMM(F) by L. minor varied between 10 and 40% after 96 h. This depuration decreased in the presence of copper possibly due to the metal toxicity.

EU regulatory aspects of resistance risk assessment.

The EPPO Standard PP 1/213(1) on resistance risk analysis is the basis for risk assessment within the authorisation process in Germany. Data for resistance risk analysis have to be provided by the applicant and risk-mitigation strategies prepared if necessary. The extent of data requirements and the intensity of the evaluation process depend mainly on the type of target organism, crop, mode of action of the active substance and the resistance history of the organism in combination with products of similar mode of action as the one being applied for. Because resistance risk is a very complicated issue during the authorisation of plant-protection products, intensive co-operation between applicants and regulators is essential prior to submission of the dossier, with due consideration of the amount of data necessary and, where required, the proposed resistance-management strategy.

What impact will EPPO's new resistance risk assessment guideline have on selection pressure in the European Union?

This paper examines practical and regulatory implications, including both costs and benefits, resulting from implementation of EPPO's new resistance risk analysis guideline. Crop-protection companies operating in Europe are preparing risk analyses and management strategies, and are monitoring for resistance. However, this is a complex and expensive process involving technical, commercial and regulatory functions. The actual cost of filling these data gaps is consequently greater than many would have anticipated. The agrochemical industry has a vested interest in managing resistance and is committed to maintaining the essential contribution of crop-protection products to sustainable agriculture. However, with increasing regulatory costs, it is important that regulatory authorities concentrate requirements for resistance risk analyses, management strategies and monitoring where these are most needed. Should these requirements be applied indiscriminately, crop-protection companies will have to consider whether or not it is economically justifiable to make these investments, or whether to remove less profitable uses from product labels. In such situations, minor crops stand to be disproportionately effected. One year after publication, it is too early to measure the guideline's impact on selection pressure. It is already clear that the guideline represents an unprecedented step forward in the regulation and harmonisation of resistance management. There are, however, valid arguments against regulation of resistance management. One of the most important criticisms that is levelled against the latter approach is that, with the best motivation and with appropriate use restrictions on labels, these actions alone do not solve the problem. It is critical that all stakeholders in the crop protection process are made aware of the importance of resistance management and of complying with prescribed strategies.

Web alert. Environmental biotechnology. Regulatory affairs.

A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Biotechnology.