Effects of a Bulking Agent on the Protein:Carbohydrate Ratio, Bioconversion, and Cost-effectiveness of a Larval Diet for Anastrepha ludens (Diptera: Tephritidae).

Although the bulking agent is categorized as 'inert', it could modify protein bioavailability and nutritional quality. In this study, the main goal was to determine if the bulking agent modified the protein:carbohydrate (P:C) ratio and bioconversion from diet biomass to larval biomass of Anastrepha ludens. The diet was altered only by modifying the type of bulking agent (corncob powder, coconut fiber, carrot fiber, oatmeal) added without changing the composition and concentration of the other components in the formulation. This allowed reclassification of the food matrices according to P:C ratios of 1:30, 1:35, 1:64, and 1:93. A food matrix with a high P:C ratio promoted a high protein and carbohydrate content in the larval hemolymph and immediately influenced the life-history traits of the larva or delayed them in the adult. The present study indicated a positive relationship between the P:G+T (glucose+trehalose) ratio in the larval hemolymph and the P:C ratio in the larval diet. Our results highlight the importance of including the optimum and real P:C ratio in whole fresh larval diets, since considering only the theoretical concentration of the formulation is not enough to understand the variation in key life-history traits. In addition, the bioconversion index should be included as an indicator of the efficacy of larval diets for mass rearing insects. A diet with high cost-effectiveness should be evaluated by taking into account flying flies as the end product of the mass rearing process to enhance operational SIT programs.

Effect of larval nutrition on the hemolymph protein composition during metamorphosis of Anastrepha obliqua.

Few studies have focused on how nutrition affects the bioavailability and investment of protein during the metamorphosis of tephritids. Our study allowed us to observe how the type and particle size of the bulking agent affected the protein composition in the hemolymph of the larva and adult of Anastrepha obliqua. Results indicated that, true protein bioavailability and protein profile was greatly modified by the bulking agent and its particle size. The physical structure of the food matrix affected the content of crude fiber (F), crude protein (P), F/P ratio, non-protein nitrogen, ammoniacal nitrogen, and α-amylase and trypsin inhibitors. Results from SDS-PAGE revealed 45 fractions with well-defined bands ranging from ~28 to ~401 kDa in larvae and adults, we found the main differences between the samples from different food matrices within the 75-100 kDa range. Hemolymph of adults from the coarse coconut fiber food matrix treatment showed a single band with a molecular weight close to 250 kDa, probably associated with a storage protein such as lipophorins. The food matrix with a coarse bulking agent had a high concentration of ammoniacal nitrogen, suggesting high microbial activity. In conclusion, the particle size of the bulking agent of the food matrix changes the bioavailability of protein in hemolymph in the adult regardless of the total concentration of protein. Also, when the particle size of the bulking agent favored the F/P ratio, higher larval density resulted in higher individual larval weight, larval yield, and adult emergence.

Big particles, best nutrition? Absorption and excretion of protein by Anastrepha obliqua larvae (Diptera: Tephritidae).

For purposes of mass-rearing fruit flies, nutrient intake through artificial diets is a challenge, artificial food content and processing should promote sufficient absorption and availability to ensure fly fitness. Bulking agents play an essential role in creating a quality diet, but its physical characteristics, such as particle size, may establish a better microenvironment for feeding and development. Currently, there is a lack of information about protein metabolism in mass-reared fruit flies. Therefore, we evaluated whether the particle size of the bulking agent affects the absorption and excretion of the proteins, as well as their effect on the life-history traits of Anastrepha obliqua. We determined the protein content of hemolymph and feces, as well as the presence of nitrogen end-products as indicators of their level of absorption in a diet elaborated with coarse and fine corn cob particles as a bulking agent. The bromatological composition showed that coarse particles increased the bioavailability and content of crude, digestible, and soluble protein for the diet and hemolymph protein of larvae alike. We found an inverse relationship between the protein content of the hemolymph and feces of the larvae. Ammonium was determined to be a product of the catabolism of proteins. Also, A. obliqua improved its development (yield and pupal weight) and fitness (adult emergence and flight ability) when larvae were reared on a coarse particle diet. In conclusion, a diet elaborated with a coarse bulking agent features increased protein bioavailability and nutritional quality, which, in turn, increases the life-history traits of A. obliqua.

Attractiveness and Sexual Competitiveness of Anastrepha obliqua Males (Diptera: Tephritidae) Fed on a Diet Enriched With Providencia rettgeri.

The West Indian fruit fly, Anastrepha obliqua (Macquart), is the second most important tephritid fruit fly in Mexico, infesting mango, hog plum and guava fruits. To control this pest, the Mexican government has implemented the use of the sterile insect technique (SIT), which involves the mass production, sterilization and release of flies. However, the A. obliqua laboratory males used in SIT are selected to a lesser extent by the wild females during competitiveness tests. The objective of this study was to compare the effects of males fed on fruit fly food enriched with Providencia rettgeri to those in males fed on food alone, assessing male mating competitiveness, capture of females using traps baited with males fed with the enriched diet and sex pheromone components. The results indicated that males fed with the diet enriched with P. rettgeri had increased mating competitiveness and captured more females in the field cage tests. However, no difference was observed in the proportion of volatile sex pheromone components identified during the calling of A. obliqua males. The results suggest the value of incorporating bacteria into the mass rearing technique of A. obliqua adults in order to improve the sexual competitiveness of males from the laboratory compared to wild males.

Mass Rearing, Quality Parameters, and Bioconversion in Drosophila suzukii (Diptera: Drosophilidae) for Sterile Insect Technique Purposes.

The spotted wing drosophila, Drosophila suzukii Matsumura, has emerged as a major invasive insect pest of small and stone fruits in both the Americas and Europe in the last decade. Females oviposit in ripening fruit, and significant economic losses can occur. Control measures are mainly associated with the use of pesticides, but the sterile insect technique (SIT), an ecologically friendly pest-specific method, could be used against this species. The objective of this study was to estimate the mass rearing, quality control parameters, and bioconversion using four artificial larval diets and their economic aspects oriented to the SIT application. Diets were based on the combination of coconut fiber, corncob powder, Brewer's and Torula yeast and were used as oviposition substrate and larval development. We found that a life cycle is completed in 10.19 ± 0.35 d and that adults live an average of 33.67 ± 0.76 d. The highest number of pupae per gram of diet and the maximum bioconversion (6%) were associated with flies developed in the coconut fiber + Brewer's yeast diet. Under our conditions, the establishment of D. suzukii required at least four generations. The use of 30 × 40 × 30 cm Plexiglas cages, each loaded with 5,000 adults and stocked with 500 g of coconut fiber and Brewer's yeast diet distributed in 15 × 5 × 10 cm plastic trays with a diet layer 3-cm thick, allows a minimum production of 84,000 pupae of D. suzukii per day.

A Novel, Low-Cost Coconut Fiber Larval Diet for Mass Rearing Anastrepha (Diptera: Tephritidae).

Mass rearing fruit flies of the genus Anastrepha for the sterile insect technique involves the use of artificial larval diets that commonly contain corn cob powder as a bulking agent. Corn cob powder varies in quality, and larval diets that contain it can be reaching at high temperatures (>34°C), which subsequently has a negative effect on pupal weight and adult survival. In addition, corn cob powder is susceptible to contamination by mycotoxins, which can inhibit larval development and cause high larval mortality. The objective of this work was to develop a low-cost artificial larval diet for mass rearing Anastrepha spp. fruit flies using coconut fiber, a nonconventional bulking agent, and to evaluate its effects on the quality parameters for A. ludens wild-type, A. ludens Tap-7 GS-strain, A. obliqua, A. serpentina, and A. striata. The coconut fiber diet resulted in an increase in the larval and pupal weight of A. ludens Tap-7 GS-strain and A. obliqua, an increase in the larval weight of A. ludens wild-type, an increase in the flight ability of A. obliqua and A. serpentina and an increase in adult eclosion in A. ludens Tap-7 GS-strain. The coconut fiber diet resulted in increased production and quality of the mass-reared flies, reduced the cost of the diet by 15 and 20% for A. ludens and A. obliqua, respectively, and led to further cost savings through reduced labor processes.

Generic Irradiation and Hot Water Phytosanitary Treatments for Mango Fruits cv. 'Ataulfo' niño Infested by Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae).

The mango fruit cv. 'Ataulfo' niño is an underdeveloped fruit that has a split on the back and a pronounced peak, and among the current total supply of commercialized mangoes cv. 'Ataulfo', approximately 2% are classified as 'Ataulfo' niño, which are strongly infested by Anastrepha ludens (Loew) (Diptera: Tephritidae) and Anastrepha obliqua Mcquart. The objective of this study was to determine the tolerance to 150-Gy generic irradiation for fruit flies of the Anastrepha genus in comparison to the development of a hot water treatment (HWT) as phytosanitary treatments for mango fruits cv. 'Ataulfo' niño infested by A. ludens and A. obliqua. The results indicated that both treatments were effective; 150-Gy irradiation and HWT at 46.3-47°C for 51 min did not result in significant effects on the external and internal color, total sugar content, firmness, pH, or weight. The sensorial quality described by the appearance, flavor, color, and odor did not show any significant differences between treatments. For both A. ludens and A. obliqua, the third larval instar was the most thermotolerant. The efficacy test was conducted with an immersion time of 47 min. A. ludens did not survive, but for A. obliqua, three larvae out of a total of 6,890 did survive and pupate. Consequently, the confirmatory test consisted of submerging mangoes infested with third-instar A. ludens and A. obliqua in water at 46.3-47°C for 51 min. In total, 67,392 A. ludens and 22,086 A. obliqua larvae were treated, and no surviving larvae were observed.

Influence of Pupation Substrate on Mass Production and Fitness of Adult Anastrepha obliqua Macquart (Diptera: Tephritidae) for Sterile Insect Technique Application.

Tephritid mass-rearing systems require an artificial substrate for pupation. Pupation substrate characteristics influence the quality of insects produced. Coconut fiber, as an alternative to the conventional pupation substrate vermiculite, was evaluated for Anastrepha obliqua Macquart (Diptera: Tephritidae) pupation behavior (pupation patterns, distribution, respiration rate, and pupal weight) and adult fitness (adult eclosion time, flight ability, and male mating competitiveness). Pupation percentage at 24 h, pupal weight, and flight ability were not significantly affected by substrate type. Adult eclosion levels of 50% were reached at 29.7 and 41.6 h for coconut fiber and vermiculite, respectively. Pupae distribution patterns differed between substrates because the larval aggregation level was reduced during the pupation process in coconut fiber. The pupae aggregation was three times greater in vermiculite than in coconut fiber. A higher respiratory rate in the last days of pupation and adult eclosion were recorded in the insects maintained in coconut fiber. Coconut fiber suitability as a pupation substrate for quality mass production of pupae and its implications for sterile insect technique are discussed.

The Effects of a Modified Hot Water Treatment on Anastrepha ludens (Diptera: Tephritidae)-Infested Mango.

The Mexican fruit fly, Anastrepha ludens (Loew), is a quarantine pest in mango (Mangifera indica L.) that can be controlled by using a hot water treatment (HWT). This treatment is normally followed by a 30-min hydrocooling (HYC) process that reduces the negative effects that the treatment has on fruit quality. However, if hot water-treated fruits are immediately immersed in water at 21 °C, the survival rate of third-instar A. ludens may be increased. The current approved treatment protocol states that if HYC is used, then treated fruit should undergo an additional 10-min HWT or on platform for 30 min before HYC. We aimed to determine the efficacy of HWT without an additional 10-min treatment before being subjected to HYC, while taking into consideration that the most important conditions are the temperature of the fruit core throughout treatment and the type of infestation, either oviposition or inoculation. Two experimental tests were conducted. Our first aim was to determine the effectiveness of HWT followed by HYC using three varieties and different size classes of mangoes ('Ataulfo' 200-375 and 401-570 g; 'Tommy Atkins' 401-500 and 501-700 g; 'Kent' 401-500 g). The four treatment combinations used to test HWT and immediate HYC at 21 °C were 1) HWT, 2) HWT/HYC, 3) HWT + 10 min/HYC, and 4) HWT/30 min on platform/HYC; an independent experiment was used for each variety. The second aim was to validate the HWT/HYC combination by performing confirmatory tests in commercial packing houses. The results showed that as long as the mango core temperature reached 45 °C during the HWT, it was not necessary to add the 10-min treatment to the HWT before HYC at 21 °C was applied. To ensure that the larvae are subjected to the HWT treatment for sufficient time to be lethal, the temperature of the fruit core throughout the treatment must be recorded.