Anastrepha Suspensa Research Articles

Insecticidal effect of new synthesized chalcone derivatives on Caribbean fruit fly, Anastrepha suspensa.

In this present study, new chalcone derivatives were synthesized from 4-aminoacetophenone, which were confirmed by spectroscopic methods. The toxic risks of chalcones to humans and the environment were investigated by a web-based platform called ADMETlab. With this program, the possible toxic effects of the compounds on liver, respiratory system, and eyes were evaluated. For the topical insecticidal activity, adult female Caribbean fruit fly, Anastrepha suspensa, was targeted. Results of the toxicity tests showed that chalcone derivatives are effective against female A.suspensa. Among the synthesized chalcones, 1-(4-cinnamoylphenyl)-3-(p-tolyl)urea (2) exhibited the greatest insecticidal activity, resulting in 73 % mortality at 100 µg/fly after 24 h, whereas other derivatives showed less than 30 % mortality. Our results demonstrate that insecticidal activity may be modulated by the presence of a certain phenyl ring in the structure of derivative 2 and, therefore, has potential for design of efficient chemicals for tephritid fruit fly management.

Sieving Fruit Pulp to Detect Immature Tephritid Fruit Flies in the Field.

Fruit flies of the Tephritidae family are among the most destructive and invasive agricultural pests in the world. Many countries undertake expensive eradication programs to eliminate incipient populations. During eradication programs, a concerted effort is made to detect larvae, as this strongly indicates a breeding population and helps establish the spatial extent of the infestation. The detection of immature life stages triggers additional control and regulatory actions to contain and prevent any further spread of the pest. Traditionally, larval detection is accomplished by cutting individual host fruits and examining them visually. This method is labor intensive, as only a limited number of fruit can be processed, and the probability of missing a larva is high. An extraction technique that combines i) mushing host fruit in a plastic bag, ii) straining pulp through a series of sieves, iii) placing retained pulp in a brown sugar water solution, and iv) collecting larvae that float to the surface was tested. The method was evaluated in Florida with field-collected guava naturally infested by Anastrepha suspensa. To mimic low populations more representative of a fruit fly eradication program, mangos and papaya in Hawaii were infested with a known, low number of Bactrocera dorsalis larvae. The applicability of the method was tested in the field on guava naturally infested by B. dorsalis to evaluate the method under conditions experienced by workers during an emergency fruit fly program. In both field and laboratory trials, mushing and sieving the pulp was more efficient (required less time) and more sensitive (more larvae found) than cutting fruit. Floating the pulp in brown sugar water solution helped detect earlier instar larvae. Mushing and sieving fruit pulp of important tephritid hosts may increase the probability of detecting larvae during emergency programs.

Risk analysis for Anastrepha suspensa (Diptera: Tephritidae) and potential areas for its biological control with Diachasmimorpha longicaudata (Hymenoptera: Braconidae) in the Americas

The Caribbean fruit fly Anastrepha suspensa (Diptera: Tephritidae) is a polyphagous pest causing economic losses in Central America, the Caribbean and South Florida. The parasitoid wasp Diachasmimorpha longicaudata (Hymenoptera: Braconidae) is the main parasitoid of A. suspensa in biological control programs. In this study, by modeling with CLIMEX software, climatically suitable areas were projected according to historical climate data. Areas with overlapping optimal climatic suitability for the joint establishment of the pest and parasitoid were mapped, indicating large areas with host presence in North, Central, and South America, with cold stress being the main climatic factor limiting distribution for both species. Tropical regions have the most potential for invasion, with optimal suitability in many areas. Through the projected distributions, this study can target quarantine strategies in areas most susceptible to invasion and establishment of the pest in each country. In addition, classical biological control with the parasitoid in areas with climatic suitability is also recommended.

Climate suitability modeling for Anastrepha suspensa (Diptera: Tephritidae): current and future invasion risk analysis.

The Caribbean fruit fly, Anastrepha suspensa (Lower, 1862) (Diptera: Tephritidae), is a pest of significant economic importance in Central America and Florida (USA). This study was carried out to examine the influence of climate change on the space-time distribution of A. suspensa on temporal and spatial scales. The CLIMEX software was used to model the current distribution and for climate change. The future distribution was performed using two global climate models (GCMs), CSIRO-Mk3.0 (CS) and MIROC-H (MR), under the emission scenarios (SRES) A2 and A1B for the years 2050, 2080, and 2100. The results indicate a low potential for global distribution of A. suspensa in all scenarios studied. However, tropical areas were identified with high climatic suitability for A. suspensa in South America, Central America, Africa, and Oceania until the end of the century. Projections of areas with climatic suitability for A. suspensa can provide helpful information to develop preventive strategies of phytosanitary management avoiding economic impacts with the introduction of the species.

Global potential distribution, climate dynamics, and essential climate variables for Anastrepha suspensa (Diptera: Tephritidae) using the CLIMEX model

Global potential distribution, climate dynamics, and essential climate variables for Anastrepha suspensa (Diptera: Tephritidae) using the CLIMEX model

Synthesis of Phthalimide Derivatives and Their Insecticidal Activity against Caribbean Fruit Fly, Anastrepha suspensa (Loew).

In this study, thirteen phthalimide derivatives were designed and synthesized. All synthesized compounds were evaluated to determine their potential for inhibitory activities against females of the Caribbean fruit fly, Anastrepha suspensa (Loew) (Diptera: Tephritidae). These efforts led to the discovery of three compounds 4a, 4c, and 4d with potent insecticidal activity (LD50 range from 0.70 to 1.91 μg/fly). Among these compounds, 4a exhibited the highest inhibitory potency with 0.70 μg/fly. In addition, in silico models indicated that compound 4a is less toxic than phthalimide and other precursors. Therefore, our results suggest that 4a has strong potential as a candidate component for developing a novel environmentally friendly insecticide for control of pest fruit flies.

Attraction and Longevity of 2- and 3-Component Food Cone Lures for the Caribbean Fruit Fly, Anastrepha suspensa (Diptera: Tephritidae)

The Caribbean fruit fly, Anastrepha suspensa (Loew) (Diptera: Tephritidae), is a quarantine pest of Citrus spp. and a production pest of guava and other specialty fruits in Florida. Effective monitoring lures and traps are needed for early pest detection and timely initiation of control measures. As part of a continued effort to identify attractive synthetic lures for the Caribbean fruit fly, we conducted field tests in Homestead, Florida to compare the efficacy and longevity of commercial 2- and 3-component cone lures (2C [ammonium acetate and putrescine], 3C [ammonium acetate, putrescine, and trimethylamine]), the current standards used by regulatory agencies, versus the traditional liquid protein bait consisting of hydrolyzed torula yeast and borax as a positive control. Additional lures were also field-aged and periodically brought into the laboratory to quantify residual chemical contents. Traps baited with the torula yeast-borax mixture captured the highest mean number of A. suspensa, and traps baited with the commercial 2C lures captured more flies than the 3C lures. Traps baited with torula yeast-borax also captured the highest number of nontarget Diptera. Captures with all three treatments were significantly biased toward females. Attractiveness of the 2C lure began to drop after 6-8 wk, and the 3C lure after 5-6 wk. Overall, these data suggest that the 2C cone lure is more attractive to A. suspensa than the 3C cone lure under field conditions in south Florida, and that the 2C lures are attractive for up to 8 wk.

Captures of Oriental Fruit Flies and Melon Flies (Diptera: Tephritidae) in Traps Baited With Torula Yeast Borax Solution Or 2- Or 3-Component Synthetic Food Cones in Hawaii

Food-based traps are an integral component of detection systems for invasive fruit fly (Diptera: Tephritidae) species which pose a serious threat to many agricultural crops. A commonly used bait is torula yeast borax solution; however, it is attractive for relatively short intervals (1–2 wk), necessitating frequent bait replacement. A dry, synthetic food bait that incorporates several components into a single matrix (or food cone) has been developed and appears to be effective for as long as 4 to 10 wk in field trials with the Mediterranean fruit fly (Ceratitis capitata [Wiedemann]) and the Caribbean fruit fly (Anastrepha suspensa [Loew]) (both Diptera: Tephritidae). Based on these results, food cones are used now in several large-scale fruit fly detection programs even though their attractiveness to other economically important tephritids, most notably Bactrocera and Zeugodacus species, has not been well studied. The goal of this study was to compare the captures of oriental fruit fly (Bactrocera dorsalis [Hendel]) and melon fly (Zeugodacus cucurbitae [Coquillett]) (both Diptera: Tephritidae) in traps baited with torula yeast borax solution with captures in traps baited with 3-component (ammonium acetate, putrescine, and trimethylamine) or 2-component (ammonium acetate and putrescine) food cones. Data from wild and released flies of both species showed that captures were significantly higher in traps baited with the torula yeast borax than those with either type of synthetic formulation. Implications of this finding for trapping programs are discussed.

Anastrepha suspensa (Caribbean fruit fly)

Anastrepha suspensa (Caribbean fruit fly)

Chemical Composition of Essential Oils from Leaves and Fruits of Juniperus foetidissima and Their Attractancy and Toxicity to Two Economically Important Tephritid Fruit Fly Species, Ceratitis capitata and Anastrepha suspensa.

The present study analyzed the chemical composition of Juniperus foetidissima Willd. essential oils (EOs) and evaluated their attractancy and toxicity to two agriculturally important tephritid fruit flies. The composition of hydrodistilled EOs obtained from leaves (JFLEO) and fruits (JFFEO) of J. foetidissima was analyzed by GC–FID and GC–MS. The main compounds were α-pinene (45%) and cedrol (18%) in the JFLEO and α-pinene (42%), α-thujone (12%), and β-thujone (25%) in the JFFEO. In behavioral bioassays of the male Mediterranean fruit fly, Ceratitis capitata (Wiedemann), both JFLEO and JFFEO showed strong attraction comparable to that observed with two positive controls, Melaleuca alternifolia and Tetradenia riparia EOs. In topical bioassays of the female Caribbean fruit fly, Anastrepha suspensa (Loew), the toxicity of JFFEO was two-fold higher than that of JFLEO, with the LD50 values being 10.46 and 22.07 µg/µL, respectively. This could be due to differences in chemical components between JFLEO and JFFEO. The JFFEO was dominated by 48% monoterpene hydrocarbons (MH) and 46% oxygenated monoterpenes (OM), while JFLEO consisted of 57% MH, 18% OM, and 20% oxygenated sesquiterpenes (OS). This is the first study to evaluate the attractancy and toxicity of J. foetidissima EOs to tephritid fruit flies. Our results indicate that JFFEO has the potential for application to the management of pest tephritid species, and further investigation is warranted.

Mitochondrial superoxide dismutase overexpression and low oxygen conditioning hormesis improve the performance of irradiated sterile males

The Sterile Insect Technique (SIT) is a successful autocidal control method that uses ionizing radiation to sterilize insects. However, irradiation in normal atmospheric conditions can be damaging for males, because irradiation generates substantial biological oxidative stress that, combined with domestication and mass-rearing conditions, may reduce sterile male sexual competitiveness and quality. In this study, biological oxidative stress and antioxidant capacity were experimentally manipulated in Anastrepha suspensa using a combination of low-oxygen conditions and transgenic overexpression of mitochondrial superoxide dismutase (SOD2) to evaluate their role in the sexual behavior and quality of irradiated males. Our results showed that SOD2 overexpression enhances irradiated insect quality and improves male competitiveness in leks. However, the improvements in mating performance were modest, as normoxia-irradiated SOD2 males exhibited only a 22% improvement in mating success compared to normoxia-irradiated wild type males. Additionally, SOD2 overexpression did not synergistically improve the mating success of males irradiated in either hypoxia or severe hypoxia. Short-term hypoxic and severe-hypoxic conditioning hormesis, per se, increased antioxidant capacity and enhanced sexual competitiveness of irradiated males relative to non-irradiated males in leks. Our study provides valuable new information that antioxidant enzymes, particularly SOD2, have potential to improve the quality and lekking performance of sterile males used in SIT programs.

Optimizing Torula Bait for Anastrepha suspensa (Diptera: Tephritidae) Trapping in the Dominican Republic

Torula yeast is the most common bait used by growers and agriculture professionals for trapping of tephritid flies in the Dominican Republic. However, the efficiency of the bait is influenced by weather conditions, aging, and contamination with undesirable microorganisms. Thus, additives such as benzalkonium chloride, a quaternary ammonium compound, have been used together with torula yeast as a bait stabilizer. This study evaluated the effect of the addition of benzalkonium chloride to torula yeast bait, and time of renewal in guava orchards for trapping of Caribbean fruit flies (Anastrepha suspensa Loew; Diptera: Tephritidae). A field study was conducted in 2 consecutive 8-wk periods between Oct 2019 and Feb 2020. Six treatments were evaluated based on the type of bait (torula yeast or torula yeast + benzalkonium chloride) and renewal frequency (weekly, biweekly, or without renewal). Treatments were arranged in a randomized complete block design with 4 replications. Data indicated that torula yeast was attractive to 85.2% and 80.2% more males and females of Caribbean fruit flies compared to torula yeast + benzalkonium chloride, respectively. Similarly, traps without renewal attracted an average of 49.8% more females than traps renewed weekly or biweekly, regardless of the bait type. Analysis of both baits showed a rapid decrease in pH of the torula yeast. The addition of benzalkonium chloride may have affected the microbial activity in the solution, leading to reduced decomposition of torula yeast + benzalkonium chloride and, therefore, reduced captures.

The hAT-family transposable element, hopper, from Bactrocera dorsalis is a functional vector for insect germline transformation

BackgroundThe hopper hAT-family transposable element isolated from the Oriental fruit fly, Bactrocera dorsalis, is distantly related to both the Drosophila hobo element and the Activator element from maize. The original 3120 bp hopperBd-Kah element isolated from the Kahuku wild-type strain was highly degenerate and appeared to have a mutated transposase and terminal sequences, while a second 3131 bp element, hopperBd-we, isolated from a white eye mutant strain had an intact transposase reading frame and terminal sequences consistent with function.ResultsThe hopperBd-we element was tested for function by its ability to mediate germline transformation in two dipteran species other than B. dorsalis. This was achieved by creating a binary vector/helper transformation system by linking the hopperBd-we transposase reading frame to a D. melanogaster hsp70 promoter for a heat-inducible transposase helper plasmid, and creating vectors marked with the D. melanogaster mini-white+ or polyubiquitin-regulated DsRed fluorescent protein markers.ConclusionsBoth vectors were successfully used to transform D. melanogaster, and the DsRed vector was also used to transform the Caribbean fruit fly, Anastrepha suspensa, indicating a wide range of hopper function in dipteran species and, potentially, non-dipteran species. This vector provides a new tool for insect genetic modification for both functional genomic analysis and the control of insect populations.

PVC Formulation of Anastrepha suspensa Pheromones Suitable for Field Studies

Abstract Tephritid flies threaten the production of fruits around the world. In the Americas, populations of the genus Anastrepha are monitored with trapping networks as part of pest management programs. Here, we report the formulation of male Anastrepha suspensa (Loew) pheromones, (±)-anastrephin and (±)-epianastrephin, into a poly(vinyl chloride) (PVC) polymer-based lure ready for trap deployment. The PVC polymer disks (100 mg) contain 10% by weight of (±)-epianastrephin and (±)-anastrephin in a naturally occurring 7:3 diastereomeric ratio, respectively. Emission of the pheromones from the disks into an airstream was evaluated as a function of abiotic environmental parameters, absolute humidity, and temperature. Kinetic data supports a diffusion-controlled mechanism of release from the matrix with first-order rate constants that decreased about 10-fold as the temperature was lowered from 30 to 15 °C. As such, the emission of volatile pheromones from the disks is suitable to last for several weeks in the field. This kinetic approach, which can be easily extended to the diffusion-controlled release of other attractants from polymeric matrices, yields laboratory predictions of the potential for environmental loss prior to conducting field bioassays.

Protein Stores Regulate When Reproductive Displays Begin in the Male Caribbean Fruit Fly.

Many animals exhibit reproductive behavior that requires expenditure of valuable nutrients. In males of many species, competitive energetically demanding displays and the development of sexual ornaments require prior accumulation of nutrient stores. Males must coordinate nutrient stores with ornament development and reproductive displays or they risk depleting their resources mid-development or mid-display, reducing their chance of mating. Males may use nutrient stores to regulate their reproductive behavior. Amino acid reserves may be important for reproduction, but the roles of amino acid stores in initiating maturation and reproductive behavior are less studied than fat stores. Insects store amino acids as hexamerin storage proteins. Many fly species use a specific hexamerin, larval serum protein 2 (LSP-2), as both a juvenile storage medium and to store protein consumed after adult eclosion. Protein stored as LSP-2 has previously been suggested to regulate reproduction in females, but no role has been proposed for LSP-2 in regulating male maturation. We use males of the Caribbean fruit fly, Anastrepha suspensa, a species with nutrient-intensive male sexual displays to test whether LSP-2 stores regulate male reproductive displays. We fed adult A. suspensa males a diet with or without protein, then assayed these males for lsp-2 transcript abundance via qRT-PCR, LSP-2 protein abundance via Western blot, and reproductive display behavior via observation. We found that adult males with ad libitum dietary protein had greater lsp-2 transcript and protein abundance, earlier sexual display behavior, and were more likely to exhibit sexual display behavior than protein-deprived adult males. We show that lsp-2 knockdown via RNAi decreases the proportion of males exhibiting reproductive displays, particularly early in the onset of reproductive behavior. Our results suggest circulating LSP-2 protein stores regulate reproductive behavior in A. suspensa males, consistent with protein stores modulating reproduction in males with expensive reproductive strategies. Our results are consistent with hexamerin storage proteins performing dual roles of protein storage and protein signaling. Our work also has substantial practical applications because tephritid flies are a pest group and the timing and expression of male reproductive displays in this group are important for control efforts using the sterile insect technique.

Bicistronic expression and differential localization of proteins in insect cells and Drosophila suzukii using picornaviral 2A peptides

Polycistronic expression systems in insects can be used for applications such as recombinant protein production in cells, enhanced transgenesis methods, and the development of novel pest-control strategies based on the sterile insect technique (SIT). Here we tested the performance of four picornaviral 2A self-cleaving peptides (TaV-2A, DrosCV-2A, FMDV 2A1/31 and FMDV 2A1/32) for the co-expression and differential subcellular targeting of two fluorescent marker proteins in cell lines (Anastrepha suspensa AsE01 and Drosophila melanogaster S2 cells). We found that all four 2A peptides showed comparable activity in cell lines, leading to the production of independent upstream and downstream proteins that were directed to the nucleus or membrane by a C-terminal nuclear localization signal (NLS) on the upstream protein and a poly-lysine/CAAX membrane anchor on the downstream protein. TaV-2A and DrosCV-2A were inserted into piggyBac constructs to create transgenic D. suzukii strains, confirming efficient ribosomal skipping in vivo. Interestingly, we found that the EGFP-CAAX protein was distributed homogeneously in the membrane whereas the DsRed-CAAX protein formed clumps and aggregates that induced extensive membrane blebbing. Accordingly, only flies expressing the DsRed-NLS and EGFP-CAAX proteins could be bred to homozygosity whereas expression of EGFP-NLS and DsRed-CAAX was lethal in the homozygous state. Our results therefore demonstrate that the 2A constructs and two novel targeting motifs are functional in D. suzukii, and that the combination of EGFP-NLS and DsRed-CAAX shows dosage-dependent lethality. These molecular elements could be further used to improve expression systems in insects and generate novel pest control strains.

Overexpression of an antioxidant enzyme improves male mating performance after stress in a lek-mating fruit fly

In many species, courtship displays are reliable signals of male quality, and current hypotheses suggest that these displays allow females to choose males with high cellular function. Environmental stressors generate excess reactive oxygen species (ROS) that impair cellular function, and thus antioxidant pathways that remove ROS are probably critical for preserving complex sexual behaviours. Here, we test the hypothesis that enhanced antioxidant activity in mitochondria preserves mating performance following oxidative stress. Using a transgenic approach, we directly manipulated mitochondrial antioxidant activity in the Caribbean fruit fly, Anastrepha suspensa, a lek-mating species with elaborate sexual displays and intense sexual selection that is also a model for sterile insect technique programmes. We generated seven transgenic lines that overexpress mitochondrial superoxide dismutase (MnSOD). Radiation is a severe oxidative stressor used to induce sterility for sterile insect programmes. After radiation treatment, two lines with intermediate MnSOD overexpression showed enhanced mating performance relative to wild-type males. These improvements in mating corresponded with reduced oxidative damage to lipids, demonstrating that MnSOD overexpression protects flies from oxidative stress at the cellular level. For lines with improved mating performance, overexpression also preserved locomotor activity, as indicated by a laboratory climbing assay. Our results show a clear link between oxidative stress, antioxidant capacity and male performance. Our work has implications for fundamentally understanding the role of antioxidants in sexual selection, and shows promise for using transgenic approaches to enhance the field performance of insects released for area-wide pest management strategies and improving performance of biological control agents in general.

Caribbean Fruit Fly Management in Florida Peaches

The Caribbean fruit fly, Anastrepha suspensa, is a member of the fly family Tephritidae, which contains some of the most destructive fruit pests in the world. This 4-page fact sheet written by Amanda Coleen Hodges and Cory Penca and published by the UF/IFAS Department of Entomology and Nematology provides information about the biology and management of the Caribbean fruit fly. http://edis.ifas.ufl.edu/in1242

CRISPR/Cas9-mediated gene editing in an exogenous transgene and an endogenous sex determination gene in the Caribbean fruit fly, Anastrepha suspensa

CRISPR/Cas9-mediated gene-editing, using injected Cas9 protein, was achieved in the Caribbean fruit fly, Anastrepha suspensa, by initially targeting an exogenous transgene, polyubiquitin-regulated EGFP (PUb-EGFP), for heritable non-homologous end-joining (NHEJ) knock-outs using an individual sgRNA. Multiple deletion mutations, ranging from two to five nts proximal to the target site, were identified phenotypically by the loss of green fluorescence in transgenic flies that were also marked with PUb-DsRed. This represented a relatively high efficiency rate of 29% for germ-line mutations. Similar conditions were then used to target an endogenous sex-determination gene, As-transformer-2 (Astra-2), using two sgRNAs that targeted independent exon sequences 671 bp apart. Somatic mutations were identified phenotypically in G0 adult flies at a frequency of 81% based upon intersexual genital morphology, expected to occur only in XX females since Astra-2 knock-outs by dsRNA do not have a phenotypic effect in XY males. Consistent with this expectation, twelve types of short indels, ranging from −15 nts to +5 nts, were identified proximal to the 5′ sgRNA-1 target site in intersexual adults. However, the 3′ sgRNA-2 target was only associated with a single 774 bp deletion extending from the sgRNA-1 target site to 100 bp downstream of the sgRNA-2 target. This is encouraging for the eventual use of dual target sites for homology-directed repair (HDR) insertions, but suggests that the sgRNA-2 target site tested may not be optimal for Astra-2 HDR modification.

Responses of Anastrepha suspensa, Diachasmimorpha longicaudata, and Sensitivity of Guava Production to Heterorhabditis bacteriophora in Fruit Fly Integrated Pest Management.

Caribbean fruit fly, also known as Caribfly or Anastrepha suspensa , is a major tephritid pest of guavas. A virulent entomopathogenic nematode (EPN) species was investigated to suppress the fruit-to-soil stages of Caribflies, which are also attacked by the koinobiont parasitoid Diachasmimorpha longicaudata in south Florida. The main objective was to develop a feasible and cost-effective EPN-application method for integrated pest management (IPM) of Caribfly to improve guava production. Naturally infested guavas were treated with increasing Heterorhabditis bacteriophora infective juvenile (IJ) concentration or rate (0, 25, 50, …, 1,600 IJs cm -2 ) in field trials to measure the optimum IJ rate and then examine sensitivity of producing guavas to inclusion of Heterorhabditis bacteriophora in Caribfly IPM plans. Relative survival of Caribfly in treatments significantly decreased with increasing IJ rate from 0 to 100 IJs cm -2 . Similarly, probability of observing large numbers of parasitoid wasps ( Diachasmimorpha longicaudata ) in EPN treatments significantly declined with increasing IJ rate (0-100 IJs cm -2 ), even though the non-target effects of Heterorhabditis bacteriophora on relative survival of Diachasmimorpha longicaudata could not be determined because of few emerging parasitoid wasps. Optimum suppression (⩾ 60%) of Caribfly was consistently achieved at 100 IJs cm -2 or 17,500 IJs fruit -1 . Profitability analysis showed that Heterorhabditis bacteriophora can be included in Caribfly IPM tactics to produce guavas. Costs of EPNs in Caribfly IPM are minimized if Heterorhabditis bacteriophora is strategically applied by spot treatment of fruit. Repayment of costs of EPN-augmentation by spot treatments appears achievable by recovering 5.71% of the annual yield losses (⩾1,963 kg ha -1 ≈ US$ 8,650 ha -1 ), which are largely due to Caribfly infestation. Hectare-wide EPN-augmentation (or broadcasting) method requires more fruit recovery than the total annual yield losses to repay its high costs. Profitability of guava production in south Florida will not be very sensitive to marginal costs of the spot treatment method, when compared to the field-wide broadcasting of Heterorhabditis bacteriophora .