Progress 07/01/18 to 02/28/19

Outputs
Target Audience:With this project, ISCA aims to strengthen and expand the commercial market for pheromone-based control of agricultural pests by introducing one of the first effective, commercially viable MD products for use in corn and other row crops, SPLAT FAW, for control of fall armyworm. Large-scale field crops, such as those targeted by SPLAT FAW, are expected to be a particularly important segment of the future global market for IPM practices relying on insect pheromones, especially as pest resistance to insecticidal Bacillus thuringiensis (Bt) proteins incorporated into transgenic crops as well as to conventional insecticides becomes more widespread. SPLAT FAW will be of significant interest to conventional and GM-crop farmers, as it will allow them to continue to use transgenic Bt crop strains despite evolving FAW resistance to those proteins. This would allow them to continue to benefit from all the advantages of these desirable bio-engineered traits, such as the control of other GM trait-susceptible moths and herbicide resistance. Use of SPLAT FAW could also reduce the need for corn, soy, and cotton farmers to keep refuge areas with non-GM crop strains, which will make it easier for farmers to grow Bt crops and alleviate refuge non-compliance issues. SPLAT FAW will also be of interest to organic farmers. Use of chemical pesticides and genetic modification to control pests in agriculture remains highly controversial in the US and abroad. Concerns about environmental damage and human health risks associated with overuse of chemical pesticides has spurred substantial growth in the organic food market, and ongoing controversy over GM crops has resulted in GMO bans in many countries and jurisdictions. SPLAT FAW will be an environmentally sustainable alternative for non-GMO farmers, who export their crops to such areas, with no environmental damage or downstream health risks to workers or consumers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?A major component of ISCA's Phase I and Phase II projects for the USDA SBIR program consists of establishing and maintaining informative and productive relationships with the various stakeholder groups affected by a particular insect management problem or opportunity, from agricultural producers and their customers, manufactures and distributors, to government officials and pest control advisors. ISCA is currently in talks with GM seed producers who are very interested in the further development of SPLAT FAW as a tool to protect crops, but more importantly, to protect and extend the commercial life of their GM products. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The objective of this project is to develop an effective SPLAT FAW formulation to disrupt the mating of FAW in corn and other row crops, thereby suppressing the pest's population in the treated area as well as the damage inflicted to the crop. Phase I studies on the development of SPLAT FAW have produced excellent results. ISCA completed successful trials on 60 ha of corn in Argentina, where FAW pressure is strong. SPLAT FAW-treated fields showed substantially less leaf damage and kernel loss from FAW larvae feeding than a control field of the same size. ISCA also formulated SPLAT FAW for tractor applications and tested the product applied in this way over 1,038 ha of corn under FAW pressure in Brazil, with similar results. SPLAT FAW formulations were registered for commercial use with US Environmental Protect Agency (EPA) in late 2018. Task 1. Formulate the optimal pheromone blend into SPLAT FAW. ISCA's SPLAT FAW was formulated as a flowable matrix with the active ingredient pheromone blend concentration carefully balanced betweenoptimal efficacy and affordability for the customers. Task 2. Determine the effect of SPLAT FAW on FAW populations in field conditions. Argentina field trial in non-Bt crops. The first field trial of ISCA's PI SPLAT FAW prototype was conducted near Tolloche, in the Salta Province of northeast Argentina, experiencing high FAW pest pressure. Sixty ha of non-Bt maize crops were assigned to receive treatment with a formulation of SPLAT FAW (0.5 kg/ha), while an additional 60 ha were designated as control. Both treatment and control fields were treated with conventional chemical insecticides at V5 and V7-V8 vegetative stages. SPLAT FAW was applied via specially designed dispensers attached to motorcycles to evenly distribute dollops of SPLAT FAW (2,500 ~200-mg point sources/ha) over the plot. Two applications of SPLAT FAW were made, the first at the V5 and the second at the V10 vegetative stage. The effect of this treatment was assessed on the basis of the number of adult male FAW captured in pheromone-baited traps, leaf damage to the crop, and kernel damage. The number of male moths captured in pheromone-baited delta traps was greatly reduced in the fields treated with SPLAT FAW, compared to those treated only with insecticide. At the VT (tassel) stage, the difference was particularly dramatic, with each trap averaging 6.8 moth captures per trap in fields treated with two applications of SPLAT FAW, compared to an average of 61.75 moths captured per trap in the control field. This indicates that nearly 90% of male moths were unable to find mates in the treated fields. In this kind of study, the traps emitting pheromones act as stand-ins for virgin females that emit the same pheromones to attract males. If the male moths can't find their way to the traps because of the abundance of synthetic pheromones emitted from SPLAT FAW dollops in the field, this indicates that they cannot find the female moths either, because of their inability to follow the pheromone plumes emitted from the delta traps. Damage to leaves was also reduced in SPLAT FAW-treated fields. At the V4 stage, before the first SPLAT treatment, the field assigned to receive that treatment had 34% of plants with levels of damage rated at Stage 3 or worse on the Davis scale, a visual assessment system designed to quantify this type of damage, compared to 28% in the control field. SPLAT treatments appeared to keep the leaf damage at the same level throughout the life of the plant (35% at Stage 3 or worse at V10), while the leaf damage more than doubled in the control field (57% at Stage 3 or worse at V10). Damage to corn ears was also reduced in crops treated with SPLAT FAW. At harvest time, sampled ears from SPLAT FAW-treated plots averaged 2.1 cm2 in damage, compared to 2.9 cm2 in the control plots. The yield losses from FAW pressure were reduced by a significant 38% by the SPLAT FAW treatment. Brazil field trial in Bt corn. ISCA conducted a second field trial of SPLAT FAW in February 2019, during which the formulation was applied by tractor in the state of Mato Grosso do Sul, Brazil. (ISCA's attractant for Noctuidae moth species made of plant volatiles) blended with the insecticide, methomyl; SPLAT FAW alone; and control (no treatment). The first treatments of SPLAT FAW only and SPLAT FAW + Noctovi + insecticide were applied by tractor during the seeding stage, a day after the corn plantings on Feb 21 and 22, 2019. Five hundred g of SPLAT FAW were applied per ha, with ~1,000 0.5-g dollops (each 15-20 mm in size) applied per ha. The second treatments were applied by during the V5 of the crop's growth, at the same rate. The performance of the SPLAT FAW mating disruption formulation was assessed using the same metrics as in the Argentina study. In addition, FAW larvae were counted on a weekly basis, with larvae classified according to life stages. Two applications of SPLAT FAW achieved a trap shutdown rate of 55-100% over the course of the trial. Male moth capture rate in SPLAT FAW-treated plots ranged from 0-0.42 moths per trap per day, compared to 0-1.42 moths per trap per day in plots treated with insecticides only. On all sampling dates on which leaf damage was reported with the exception of V2 and V7, lower rates of this damage were observed in SPLAT FAW-treated plots than in insecticide-treated plots. At four sampling time points--those occurring at the V4, V5, V6, and V7 growth stages of the Bt crop--rates of leaf damage exceeded the established threshold above which insecticide sprays are deemed necessary. In contrast, the damage threshold was exceeded six times on plots that did not receive treatment with SPLAT FAW (insecticide only), during V4, V5, V6, V7, V8, and V9 sampling dates. SPLAT FAW-treated plots showed lower rates of FAW-infested plants on four out of seven sampling dates on which evidence of infestation was found: V3, V4, V6, and V8. Percentage of infested plants ranged from 0-45% in crops treated with SPLAT FAW and insecticides, peaking at the V7 growth stage, versus 0-75% in crops treated with insecticides alone, peaking at V8. Lower total numbers of larvae, accumulated between V2 and V10, were detected in Bt crops treated with SPLAT FAW + insecticides than in the same crop treated only with insecticides. Final assessment of FAW-related damage to corn ears harvested from the Bt crop fields subjected to the two experimental treatments are planned for [target date]. These results will be submitted in an addendum to the PI final report. Task 3. Involve stakeholders in problem identification and implementation of results. In addition to continuing outreach and engagement with the row crop growers who will ultimately purchase and use SPLAT FAW (See response to later question re: Dissemination of project results to communities of interest.), ISCA has achieved a major advance toward successful commercialization of this MD product. In November 2018, ISCA obtained EPA registrations for its blend of semiochemical AIs and three SPLAT FAW formulations containing different concentrations of these AIs mixed with our SPLAT-slow release matrix, which is made of environmentally safe food grade materials, including wax, vegetable oils, water, and emulsifiers. The AIs are environmentally safe sex pheromone constituents (Z)-9-tetradecenyl acetate, (Z)-11-hexadecenyl acetate, and (Z)-7-dodecenyl acetate. Obtaining EPA registrations is a crucial, legally required milestone on the path toward commercialization, and the acquisition of separate registrations for the pheromone blend and three field formulations now gives ISCA great flexibility in meeting grower needs developing specific products for marketing.

Publications