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Background: Control of Zeugodacus cucurbitae, a serious agricultural pest worldwide, often includes or is dependent on the use of spinosad-based insecticides. This is especially the case in Hawaii, where GF-120, a protein bait containing spinosad as the active ingredient, has been in use as a key integrated pest management (IPM) tool against this Tephritid for the last two decades. Here, we report on resistance to spinosad [resistance ratios (RRs) and median lethal concentration (LC 50 )] in Hawaii's populations of Z. cucurbitae., Results: High resistance was found in populations from three farms on Oahu (RR = 102-303; LC 50  = 191-567 mg L -1 ) and in a population from Maui (RR = 8.50; LC 50  = 15.9 mg L -1 ). These will be problematic for control given that the most concentrated dilution ratio on the GF-120 label is 96 mg L -1 of spinosad (1 part GF-120 to 1.5 parts water). Background resistance in a naïve wild population from the Island of Hawaii (RR = 2.73; LC 50  = 5.1 mg L -1 ) was relatively low compared with a spinosad-susceptible laboratory colony (LC 50  = 1.87 mg L -1 ). Resistance in the three Oahu and one Maui populations declined over generations in the absence of spinosad but remained elevated in some cases. Moreover, melon flies collected from one of the Oahu farms 1 year after the cessation of spinosad use revealed high persistence of resistance., Conclusion: Compared with a 2008 survey of spinosad resistance, our findings indicate a 34-fold increase in resistance on one of the Oahu farms over 9 years. The evolution and persistence of high levels of resistance to spinosad in Z. cucurbitae in Hawaii highlights the need for alternative control tactics, particularly rotation of active ingredients. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Spinosad is a natural insecticide with desirable qualities, and it is widely used as an alternative to organophosphates for control of pests such as the melon fly, Bactrocera cucurbitae (Coquillett). To monitor the potential for development of resistance, information about the current levels of tolerance to spinosad in melon fly populations were established in this study. Spinosad tolerance bioassays were conducted using both topical applications and feeding methods on flies from field populations with extensive exposure to spinosad as well as from collections with little or no prior exposure. Increased levels of resistance were observed in flies from the field populations. Also, higher dosages were generally required to achieve specific levels of mortality using topical applications compared to the feeding method, but these levels were all lower than those used for many organophosphate-based food lures. Our information is important for maintaining effective programs for melon fly management using spinosad.

Determination of physiological state in insects is useful in furthering the understanding of how insect behavior changes with age. Central to this determination is the identification of characters that allow assessment of physiological age. While non-destructive measures produce the most desired outcomes, internal markers may be more diagnostic and reliable. In this study, key morphological characters during previtellogenesis through vitellogenesis and ovulation were assessed as markers to determine physiological states of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). Ovary length and width, ovarian index (length × width), and egg load of laboratory-reared B. dorsalis females recorded daily from eclosion up to 80 days old suggested significant differences in the ovarian index and egg load between females from each oogenesis stage. Parity status determined by the presence of follicular relics was found to provide high-accuracy classifications for B. dorsalis females. The presence of follicular relics with distinct morphological features provides a reliable identification tool to determine the physiological state of wild female oriental fruit fly. The potential applications of this technique to identify the physiological age of female fruit flies to study behavioral attributes in their natural habitat, and also the potential applications in relation to field control, are discussed.

The effectiveness of foliar applications of protein baits against pestiferous fruit flies (Tephritidae) can be adversely affected by a rapid loss of attractive volatile compounds and by rainfall due to the high water solubility of the baits. In a large coffee, Coffea arabica L., plantation in Hawaii with high and low populations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), the relative attractiveness of GF-120 NF Naturalyte Fruit Fly Bait as either a 40% (vol:vol) spray solution (= GF-120 NF) or as a formulated proprietary amorphous polymer matrix (= GF-120 APM) was compared. The GF-120 APM formulations contained either, 25, 50, or 75% of GF-120 NF (wt:wt). All baits were tested in association with visually attractive yellow bait stations as a way of standardizing the evaluations. With both high and low C. capitata populations, significantly more females were attracted to the fresh sprayed GF-120 NF than to any of the three fresh GF-120 APM formulations. The attractiveness of GF-120 sprayed decreased significantly after 1 wk, whereas 1-wk-old GF-120 APM formulations were as attractive as similar fresh formulations. GF-120 APM 75% aged for 3 wk outperformed similarly-aged sprayed GF-120 NF with comparatively high C. capitata populations. With low populations, both GF-120 APM 75% and GF-120 APM 50% aged for 2 wk outperformed the similarly aged sprayed GF-120 NF. Combined findings indicate that APM mixed with either 50 or 75% GF-120 applied to bait stations can be attractive to female C. capitata for up to 3 wk longer than the standard sprayed GF-120 NF.

Ammonia-releasing substances are known to play an important role in fruit fly (Diptera: Tephritidae) attraction to food sources, and this information has been exploited for the development of effective synthetic food-based lures and insecticidal baits. In field studies conducted in Hawaii, we examined the behavioural response of wild female oriental fruit fly (Bactrocera dorsalis (Hendel)), melon fly (B. cucurbitae (Coquillett)), and Mediterranean fruit fly (Ceratitis capitata (Wiedemann)) to spinosad-based GF-120 NF Naturalyte Fruit Fly Bait(©) formulated to contain either 0, 1 or 2% ammonium acetate. Use of visually-attractive yellow bait stations for bait application in the field allowed for proper comparisons among bait formulations. Field cage tests were also conducted to investigate, using a comparative behavioural approach, the effects of female age and protein starvation on the subsequent response of F1 generation B. cucurbitae and B. dorsalis to the same three bait formulations that were evaluated in the field. Our field results indicate a significant positive effect of the presence, regardless of amount, of AA in GF-120 for B. dorsalis and B. cucurbitae. For C. capitata, there was a significant positive linear relationship between the relative amounts of AA in bait and female response. GF-120 with no AA was significantly more attractive to female C. capitata, but not to female B. dorsalis or B. cucurbitae, than the control treatment. Our field cage results indicate that the effects of varying amounts of AA present in GF-120 can be modulated by the physiological stage of the female flies and that the response of female B. cucurbitae to GF-120 was consistently greater than that of B. dorsalis over the various ages and levels of protein starvation regimes evaluated. Results are discussed in light of their applications for effective fruit fly suppression.

Studies were conducted in Hawaii to measure attraction of male melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), to SPLAT-Cue-Lure (C-L) and SPLAT-Melo-Lure (M-L) (raspberry ketone formate). Direct field comparisons of SPLAT-C-L and SPLAT-M-L at low (5%) and high (20%) concentrations indicated few differences in attraction over a 15-wk period. Subsequently, only SPLAT-Spinosad-C-L (5%) was compared with Min-U-Gel C-L with naled (standard used in California) in weathering studies. Treatments were weathered for 1, 2, 4, and 8 wk in Riverside, CA, and shipped to Hawaii for attraction/toxicity tests under field and semifield conditions by using released males of controlled ages, and for feeding tests in the laboratory. In terms of attraction, SPLAT-Spinosad-C-L compared favorably to, or outperformed the current standard of Min-U-Gel-C-L with naled. In terms of toxicity, the cumulative 24-h mortality did not differ between the two insecticide-containing C-L treatments in field cage studies after 8 wk. However, in feeding studies in which individual males were exposed for 5 min to the different C-L treatments after 4 wk of weathering, SPLAT-Spinosad-C-L demonstrated reduced mortality compared with the Min-U-Gel-C-L with naled, suggesting reduced persistence of the spinosad material. Spinosad has low contact toxicity and when mixed with SPLAT and C-L offers a reduced risk alternative for control of B. cucurbitae and related C-L-responding species, without many of the negative effects to humans and nontargets of broad-spectrum contact poisons such as naled.

Methyl eugenol (ME) and cue-lure (C-L) traps with solid lure dispensers were deployed in areas with low and high populations of oriental fruit fly, Bactrocera dorsalis (Hendel), and melon fly, Bactrocera cucurbitae (Coquillett), respectively. In low-density areas, standard Jackson traps or Hawaii Fruit Fly Areawide Pest Management (AWPM) traps with FT Mallet ME wafers impregnated with dimethyl dichloro-vinyl phosphate (DDVP) or AWPM traps with Scentry ME cones and vapor tape performed equally as well as standard Jackson traps with liquid ME/C-L and naled. Standard Jackson traps or AWPM traps with FT Mallet C-L wafers impregnated with DDVP or AWPM traps with Scentry C-L plugs with vapor tape performed equally as well as standard Jackson traps with a lure-naled solution. In high density areas, captures with traps containing FT Mallet wafers (ME and C-L) outperformed AWPM traps with Scentry cones and plugs (ME and C-L) with DDVP insecticidal strips over a 6-mo period. Captures of B. dorsalis and B. cucurbitae with wafers containing both ME and raspberry ketone (FT Mallet MC) were equivalent to those containing separate lures. From a worker safety and convenience standpoint, FT Mallet ME and C-L wafers with DDVP or Scentry plugs, with or without DDVP vapor tape, are more convenient and safer to handle than standard liquid insecticide formulations used for monitoring and male annihilation programs in Hawaii, and for detections traps used on the U.S. mainland. Furthermore, the FT Mallet MC wafer might be used in a single trap in place of two separate traps for detection of both ME and C-L responding fruit flies.

The United States Department of Agriculture's Agricultural Research Service initiated an area-wide fruit fly management program in Hawaii in 2000. The first demonstration site was established in Kamuela, Hawaii, USA. This paper documents suppression of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in a 40 km2 area containing urban, rural and agricultural zones during a 6 year period. The suppression techniques included sanitation, GF-120 NF Naturalyte Fruit Fly Bait sprays, male annihilation, Biolure traps, and parasitoids against C. capitata and B. dorsalis. In addition, small numbers of sterile males were released against B. dorsalis. Substantial reductions in fruit infestation levels were achieved for both species (90.7 and 60.7% for C. capitata and B. dorsalis, respectively) throughout the treatment period. Fruit fly captures in the 40 km2 treatment area were significantly lower during the 6 year period than those recorded in three non-treated areas. The strategy of combining suppression techniques in an area-wide approach is discussed.

Bait stations represent an environmentally friendly attract-and-kill approach to fruit fly population suppression. Recently a novel, visually attractive, rain-fast bait station was developed in Hawaii for potential use against multiple species of pestiferous fruit flies. Here, we compared the efficacy of GF-120 NF Naturalyte Fruit Fly Bait applied either as foliar sprays or onto bait stations in reducing female oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), population density and level of fruit infestation in commercial papaya orchards in Hawaii. Trapping and infestation data were used as indicators of the effectiveness of the two bait application methods. For the first 10 weeks of the study, captures of female B. dorsalis in monitoring traps were significantly greater in control plots than in plots treated with foliar sprays or bait stations. Six weeks after the first bait spray, incidence of infestation (i.e. number of fruit with one or more B. dorsalis larvae) of quarter to half-ripe papaya fruit was reduced by 71.4% and 63.1% for plots with bait stations and foliar sprays, respectively, as compared to control plots. Twelve weeks after first spray, incidence of infestation was reduced by only 54.5% and 45.4% for plots with bait stations and foliar sprays, respectively, as compared to control plots. About 42% less GF-120 was used in orchard plots with bait stations compared to those subject to foliar sprays. The impact of field sanitation on the outcome is also discussed. The results indicate that bait stations can provide a simple, efficient, and economical method of applying insecticidal baits to control fruit flies and a safer alternative to foliar sprays.

The efficacy of GF-120 NF Naturalyte Fruit Fly Bait in combination with field sanitation was assessed as a control for female oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in papaya (Carica papaya L.) orchards in Hawaii. Three different bait spray regimes were evaluated: every row (high use of the bait), every fifth row (moderate use), and every 10th row (low use). Orchard plots in which no bait was applied served as controls. For five of the seven biweekly periods that followed the first bait spray, trapping data revealed significantly fewer female B. dorsalis captured in plots subject to high and moderate bait use than in control plots. Differences in incidence of infestation among treatments were detected only by the third (12 wk after first spray) fruit sampling with significantly fewer infested one-fourth to one-half ripe papaya fruit in plots subject to high and moderate bait use than in control plots. Parasitism rates by Fopius arisanus (Sonan) (Hymenoptera: Braconidae) were not negatively affected by bait application. Results indicate that foliar applications of GF-120 NF Naturalyte Fruit Fly Bait either to all rows (every other tree), or to every fifth row (every tree) in combination with good sanitation can effectively reduce infestation by B. dorsalis in papaya orchards in Hawaii.

Methyl eugenol (ME) and cue-lure (C-L) traps to detect tephritid flies on the U.S. mainland were tested with and without insecticides under Hawaiian weather conditions against small populations of oriental fruit fly, Bactrocera dorsalis (Hendel) and melon fly, Bactrocera cucurbitae (Coquillett), respectively. In comparative tests, standard Jackson traps with naled and the Hawaii fruit fly areawide pest management (AWPM) trap with 2,2-dichorovinyl dimethyl phosphate (DDVP) insecticidal strips outperformed traps without an insecticide. Addition of the reduced risk insecticide spinosad did not increase trap capture significantly compared with Jackson traps without an insecticide. Captures in AWPM traps with DDVP compared favorably with those for the Jackson trap with liquid naled (the Florida standard). In subsequent tests, captures with solid Farma Tech wafer dispensers with ME or C-L and DDVP placed inside Jackson and AWPM traps were equal to those for a Jackson trap with naled, currently used for detection of ME and C-L responding fruit flies in Florida. Farma Tech ME and C-L wafers with DDVP would be more convenient and safer to handle than current liquid insecticide formulations (e.g., naled) used for detection programs in Florida.

Attraction and feeding responses of oriental fruit fly, Bactrocera dorsalis (Hendel), and melon fly, Bactrocera cucurbitae (Coquillett), were determined for different protein baits. In separate choice attraction assays for each species, significantly more flies arrived at stations with bait than water, but no differences existed among baits of GF-120 Fruit Fly Bait, GF-120 NF Naturalyte Fruit Fly Bait, Provesta 621 autolyzed yeast extract, and Mazoferm E802. In comparison with B. dorsalis, B. cucurbitae had 2.8 times more responders and a 4.8 times better discrimination between baits and water. In a second attraction assay with only B. dorsalis, volume of bait was negatively correlated to numbers of flies alighting on the bait. Feeding assays for both species demonstrated that time spent feeding and duration on a leaf were both significantly affected by bait type. B. dorsalis fed the longest on Provesta 621, with significantly less feeding on the other baits, and with all baits resulting in more feeding than water. The longest feeding times for B. cucurbitae resulted with Mazoferm E802 and Provesta 621, and all baits except GF-120 NF resulted in eliciting a significantly longer feeding duration than water. In separate toxicology assays for each species, significantly higher mortality resulted from bait formulations containing spinosad compared with blank baits, but no differences existed between GF-120 and GF-120 NF formulations. The differences are discussed between the two Bactrocera species primarily in regard to bait preference, extent of response, and previous work on laboratory flies.

Six to nine populations of the diamondback moth, Plutella xylostella (L.), were collected annually from fields of crucifer vegetables in the United States and Mexico from 2001 to 2004 for baseline susceptibility tests and resistance monitoring to spinosad, indoxacarb, and emamectin benzoate. A discriminating concentration for resistance monitoring to indoxacarb and emamectin benzoate was determined based on baseline data in 2001 and was used in the diagnostic assay for each population in 2002-2004 together with a discriminating concentration for spinosad determined previously. Most populations were susceptible to all three insecticides, but a population from Hawaii in 2003 showed high levels of resistance to indoxacarb. Instances of resistance to spinosad occurred in Hawaii (2000), Georgia (2001), and California (2002) as a consequence of a few years of extensive applications in each region. The collaborative monitoring program between university and industry scientists we discuss in this article has provided useful information to both parties as well as growers who use the products. These studies provide a baseline for developing a more effective resistance management program for diamondback moth.

Fourteen populations of the diamondback moth, Plutella xylostella (L.), were collected from fields of crucifer vegetables in the United States, Mexico, and Thailand in 1999 and 2000 for susceptibility tests with spinosad. Most populations were susceptible to spinosad and similar to earlier baseline values, but populations from Thailand and Hawaii showed high levels of tolerance. A statewide survey in Hawaii in 2000 and 2001 indicated resistance problems on several islands. One colony collected in October 2000 from Pearl City, HI, was subjected to further selection pressure, using spinosad in the laboratory, and then was used as the resistant strain (Pearl-Sel) for other tests. Spray tests using the recommended field rates of spinosad on potted broccoli plants in the greenhouse confirmed that field control failures due to resistance were possible in the areas of these collections. Analysis of probit lines from F1 reciprocal crosses between the Pearl-Sel and S strain indicated that resistance to spinosad was inherited autosomally and was incompletely recessive. A direct test of monogenic inheritance based on the F1 x Pearl-Sel backcrosses suggested that resistance to spinosad was probably controlled by one locus. The synergists S,S,S-tributyl phosphorotrithioate and piperonyl butoxide did not enhance the toxicity of spinosad to the resistant colony, indicating metabolic mediated detoxification was probably not responsible for the spinosad resistance. Two field colonies in Hawaii that were resistant to spinosad were not cross-resistant to emamectin benzoate or indoxacarb. Resistance developed in Hawaii due to the continuous cultivation of crucifers in which as many as 50 applications of spinosad per year may have been made to a common population of P. xylostella in sequential plantings, although each grower might have used the labeled restrictions for resistance management. Resistance management strategies will need to address such cropping and pest management practices.