Performing Department
ApexBait Technologies
Non Technical Summary
The goal of this USDA NIFA SBIR Phase I project is to develop Apex-FLB (Fly Liquid Bait), an inexpensive and highly effective liquid bait matrix, to attract and kill blood-feeding stable fliesin livestock production systems while remaining safe for non-target organisms. The stable fly is one of the most damaging pests, afflicting livestock animals in the USA and around the world. The USA, as the largest producer of beef (>12 million tons/yr.) and cow milk (>99 million tons/yr.) in the world, suffers an estimated annual loss of $402 million in dairy and $2.01 billion in the beef industries in the absence of an effective stable fly control solution. In addition, exotic zoo animals, wild animals, and humans also suffer from stable fly bites, extending the scope of the problem and the market opportunity for an effective stable fly control solution. Current methods employed for stable fly management are limited, primarily focused on sanitation, insecticide treatment, and trap deployment. The development of resistance against commonly used chemical insecticides, environmental concerns, and failure of vacuum fly traps in controlling stable flies has created an urgent need for novel, effective, and environmentally safe stable fly control technologies. Apex Bait Technologies, Inc. will develop a novel blood substitute liquid bait (Apex-FLB) with unique attractants, phagostimulants, and a chemical insecticide that will ensure the attraction and direct consumption of toxicants by stable flies, causing complete mortality and avoiding resistance development. The objective of this Phase I project is to formulate and optimize the efficiency of the liquid bait in semi-field and field trials. In Phase I research we will gather extensive data on the effectiveness of Apex-FLB formulation in stable fly control under diverse environments. The development of Apex-FLB attract and kill liquid bait matrix will build on recent breakthroughs achieved by Apex Bait Technologies in the development of a blood substitute liquid bait matrix and on decade-long experience in the development and commercialization of bait matrices for urban and veterinary pests.
Animal Health Component
60%
Research Effort Categories
Basic
25%
Applied
60%
Developmental
15%
Goals / Objectives
The goal of this USDA NIFA SBIR Phase I project is to develop Apex-FLB (Fly Liquid Bait), an inexpensive and highly effective liquid bait matrix, to attract and kill stable flies (Stomoxys calcitrans) in livestock production systems while remaining safe for non-target organisms.The objectives essential to accomplish the above-mentioned goalare-Objective 1. Optimize the addition of stable fly attractant and phagostimulants in liquid bait base (0-3 months).Objective 2. Optimize the liquid bait for the delivery of active ingredients to stable flies (2-4 months).Objective 3: Determine the feasibility of liquid bait for the control of stable flies in semi-field and field trials (4-8 months)
Project Methods
Task 1: Testing the effects of chemical attractant incorporation in bait matrix base on the stable fly attraction.Methods: Behavioral assays will be conducted to test whether the incorporation of chemical attractants (1-hexanol and 2-phenyl ethanol) in the Apex-FLB base enhances the attraction of stable flies to the Apex-FLB base. The attraction of stable flies will be measured in terms of the numbers of stable flies feeding on the Apex-FLB base. In brief, A known number of stable flies (50-200) of mixed age and sex will be released into the experimental arena (2ft x 2ft x 2ft cage for a short distance and 10ft x 7ft x 6ft tent arena for long-distance attraction) and will be allowed to acclimate before the start of choice assays. After one day of acclimatization, stable flies will be given a choice of pre-weighed bait base (Apex-FLB base) with and without the attractant (5μg). The stable fly attraction will be measured by counting the number of stable flies caught in the funnel trap with bait base and attractant incorporated bait base within the first 30 minutes. We will also measure the feeding rate by dividing the amount of bait consumed (initial weight of bait - final weight of bait) by the number of stable flies in the funnel trap. This will be done to evaluate the effect of attractant incorporation on the palatability of bait. The collected data will be analyzed using a paired t-test or Wilcoxon signed-rank test for determining the significant effect. Similar behavioral bioassays will be used to determine the optimal concentrations of the1-hexanol and 2-phenyl ethanol that produces the highest attraction of stable flies when mixed with the bait base. Follow up tests will be performed to test if combining the 1-hexanol and 2-phenyl lead to a synergistic effect on stable fly attraction. We anticipate that attractant incorporation in the Apex-FLB base will improve the stable fly trapping >50% compared to the base alone. Once we finalize the optimal concentration of chemical attractants that generate the highest stable fly attraction, we will use it for the second task of this objective.Task 2: Testing the effects of phagostimulant addition in bait matrices on the bait acceptance by stable flies.Methods: Feeding bioassays will be conducted to evaluate the effect of phagostimulant addition to the attractant-incorporated Apex-FLB base on stable fly feeding. In brief, A known number of stable flies (100) of mixed sex and age will be given a choice of pre-weighed, attractant incorporated Apex-FLB base with and without phagostimulant in individual funnel trap in a 2ft x 2ft x 2ft cage. After an hour of feeding, we will measure the total bait consumption (initial weight of bait -final weight of bait) and feeding rate (bait consumption/number of stable flies in the funnel trap). The effect of phagostimulant on stable fly feeding will be determined by analyzing total consumption and feeding rate using standard statistical tests (paired t-test). We anticipate that phagostimulant addition will improve the stable fly feeding by >50%.Task 3:Optimize the liquid bait for the delivery of active ingredients to stable flies.Various concentrations of boric acid (1%, 2.5%, and 5%) will be added to attractant and phagostimulant incorporated liquid bait obtained from objective 1. Stable flies of mixed age and sex will be allowed to feed on chemical insecticide laced bait in a similar bioassay as described above for phagostimulant bioassays (task-2 methods). The efficacy of boric acid in liquid bait will be determined by analyzing mortality over 1-3days using standard statistical tests (Mantel-Cox Log-rank test). Our results from the preliminary experiment showed that a 1% concentration of boric acid in blood caused >75% mortality in stable flies after one day of feeding (Figure 2). Therefore, we anticipate boric acid in bait matrices to elicit >75% mortality within a day and >95% mortality at low, medium, and high concentrations in 3 days.Task 4: Determine the feasibility of liquid bait for the control of stable flies in semi-field and field trials.For semi-field and field tests, the prototype liquid bait formulations will be loaded in bait stations consisting of a 5ml conical plastic vial which will be plugged with a 3 cm segment of cotton wick. The semi-field trials will be conducted at our company headquarters in Santa Clara, CA in a large indoor arena of 10ft x 7ft x 6ft. A batch of ~500 stable flies of mixed sex and age will be released into the testing arena and allowed to acclimate for 24 hours before the start of the experiment. After a day of acclimatization, the stable flies will be exposed to one of the three prototype liquid bait formulations deployed using the bait station. We predict that our prototype liquid bait will result in >95% mortality of stable flies within 3 days and the complete collapse of the stable fly population over a week. The comparative efficacy of prototype liquid bait formulations will be determined by analyzing stable fly mortality over 1-7 days using the Mantel-Cox Log-rank test.To verify if the results of semi-field trials translate in field trials, the prototype liquid bait formulations that perform better in semi-field trials will be tested in field trials. For field trials, we have established a collaboration with Dr. Junwei J Zhu, USDA-ARS, and adjunct professor at the University of Nebraska-Lincoln. The prototype liquid baits will be sent to Dr. Zhu for field trials at the dairy and beef cattle farms and feedlot area in a Nebraska cattle research farm. The field trials will be performed over three-four months. Our team will also compare the efficacy of our prototype liquid bait with currently available commercial products such as Olsen and Nzitraps in the feedlot in a Nebraska cattle research farm and expects to achieve at least 30% and optimally >50% higher control of stable fly relative to other market products.