Detection of Ochyromera ligustri (Coleoptera: Curculionidae: Curculioninae) in Ligustrum spp. (Oleaceae) Using Newly Developed PCR Primers.

Ligustrum spp. (Oleaceae) have become invasive species in the US and negatively affect native plant diversity and richness in forests. Ochyromera ligustri (Coleoptera: Curculionidae) is considered a potential biological control agent in the US because adults feed on the foliage and larvae are seed-feeders of Ligustrum spp. To discover the relationships between O. ligustri and Ligustrum spp., fruit dissections or rearing and field observations are required. In the current research project, novel PCR primers were developed to rapidly detect the DNA of O. ligustri in molecular analyses without rearing and observation. The developed PCR primers worked even with 0.01 ng of DNA and did not amplify the DNA of the other five curculionid species tested. When the novel primers were tested with three Ligustrum spp. species common in the southeastern US, the DNA of O. ligustri was detected from all three species. We expect that the novel primers will be utilized to find out the presence and impact of O. ligustri on Ligustrum spp rapidly and accurately.

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.

Coconut oil derived five-component synthetic oviposition deterrent for oriental fruit fly, Bactrocera dorsalis.

BACKGROUND: Bactrocera dorsalis, oriental fruit fly (OFF), is one of the most destructive agricultural pests. Although bait sprays can effectively control OFF, resistance development has been a concern. We evaluated the oviposition deterrent activity of coconut free fatty acids (CFFA), a mixture of eight coconut oil-derived fatty acids known to repel hematophagous insects and deter their feeding and oviposition, against OFF females. RESULTS: In laboratory 72-h two-choice assays using guava-juice infused-agar as an oviposition substrate, CFFA deterred OFF oviposition in a dose-dependent manner with the greatest reduction of 87% at 20 mg dose compared to the control. When the eight CFFA components were tested individually, four compounds (caprylic, capric, oleic, and linoleic acids) significantly reduced OFF oviposition ('negative-compounds'), two (lauric and myristic acids) had no effect ('neutral-compounds'), and two (palmitic and stearic acids) stimulated OFF oviposition ('positive-compounds'). In two-choice tests, the 'negative-compounds' blend failed to elicit the same level of oviposition reduction as CFFA at equivalent concentrations found in CFFA. Adding the two 'neutral-compounds' recovered the oviposition deterrence similar to CFFA. Subsequent subtraction tests showed that four 'negative-compounds' plus lauric acid was as effective as CFFA in reducing OFF oviposition in guava-juice agar. This five-component key-deterrent blend also reduced OFF oviposition by 95 and 72% on papaya and tomato fruit, respectively. CONCLUSION: CFFA acts as an oviposition deterrent for OFF. Given that CFFA compounds are generally regarded as safe for humans and the environment, CFFA and its bioactive components have potential use in behavioral control strategies against OFF. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Sesame as an Alternative Host Plant to Establish and Retain Predatory Mirids in Open-Field Tomatoes.

The silverleaf whitefly (Bemisia tabaci) and the South America tomato pinworm (Tuta absoluta) are two of the most destructive pests of tomato. Open-field tomato production frequently relies on chemical treatments, which has been shown to lead to pesticide resistance. The integration of biological control using predatory mirid bugs is an effective alternative method for managing these pests. However, methods to establish and maintain populations of zoophytophagous mirids are not adequately described. We explored the potential use of two mirids naturally occurring in Florida, Nesidiocoris tenuis and Macrolophus praeclarus. We conducted 6 field experiments over 4 consecutive years to develop a strategy to maintain the mirids. Pre-plant inoculation of tomato plants did not lead to their establishment, likely due to the low prevalence of prey. We explored the use of sesame (Sesamum indicum) to retain the mirids. Intercropping sesame maintained the populations of N. tenuis throughout the duration of the crop. Macrolophus praeclarus never established in any of the open-field experiments. Nesidiocoris tenuis damage was minimal (<1 necrotic ring/plant) and mirid damage was reduced in the presence of sesame. Our results show that intercropping sesame may provide a means to utilize mirids to manage B. tabaci, an established pest, and provide options to tomato growers should T. absoluta invade USA.

Fermenting Bread Dough as a Cheap, Effective, Nontoxic, and Generic Attractant for Pest Snails and Slugs.

Invasive slugs and snails are among the most damaging pests of agriculture in temperate and tropical regions of the world. Control options, however, are limited and there is a heavy reliance on chemical molluscicides of variable efficacy. There is an ongoing need to improve management methods. Here, we show that a simple fermenting bread dough formulation (flour, water, and yeast) was effective in attracting pest mollusk species in laboratory tests, and in multiple replicated field trials in Hawaii, Oregon, and Montana. The dough attracted substantially more terrestrial pest gastropods, including invasive species of major economic importance such as Cornu aspersum, Deroceras reticulatum, Ambigolimax valentianus, Xerolenta obvia, Lissachatina fulica, and Parmarion martensi, than water controls. The dough remained attractive for at least 8 days and was significantly more attractive than a widely used metaldehyde-based bait, Deadline® M-Ps™. Thus, fermenting bread dough represents a nontoxic, generic, and effective tool to aid in managing pest gastropod infestations, either using baited traps or in attract-and-kill approaches. Given its simplicity, low cost, and the ready availability of its ingredients, the dough also has potential to be used in developing countries where access to commercial molluscicide baits is limited by cost.

Biological traits of the predatory mirid Macrolophus praeclarus, a candidate biocontrol agent for the Neotropical region.

The predatory mirid Macrolophus praeclarus is widely distributed throughout the Americas, and is reported to prey upon several horticultural pest species. However, little is known about its biology, thermal requirements, crop odour preferences, phytophagy, and capability to induce defensive responses in plants. When five temperatures studied (20, 25, 30, 33 and 35°C) were tested and Ephestia kuehniella was used as prey, the developmental time from egg to adult on tomato, was longest at 20°C (56.3 d) and shortest at 33°C (22.7 d). The ability of nymphs to develop to adults decreased as the temperature increased, with the highest number of nymphs reaching the adult stage at 20°C (78.0%) and lowest at 35°C (0%). The lower and upper developmental thresholds were estimated at 11.2° and 35.3°C, respectively. The maximum developmental rate occurred at 31.7°C and the thermal constant was 454.0 ± 8.1 degree days. The highest predation rate of E. kuehniella eggs was obtained at 30°C. In Y-tube olfactory choice tests, M. praeclarus selected tomato, sweet pepper and eggplant odours more frequently than no plant control treatment. Macrolophus praeclarus feeding did not damage tomato plants compared to another zoophytophagous mirid, Nesidiocoris tenuis, which caused necrotic rings. The phytophagy of M. praeclarus induced defensive responses in tomato plants through the upregulation of the jasmonic acid metabolic pathway. The implications of the findings for using M. praeclarus in tomato biological control programmes in the Americas are discussed.

Biological Control Potential and Drawbacks of Three Zoophytophagous Mirid Predators against Bemisia tabaci in the United States.

Miridae (Hemiptera) of the tribe Dicyphini are important zoophytophagous predators use to control pest arthropods in vegetable crops. However, the risk that their herbivory may cause economic damage could hinder their application as useful biocontrol agents and may limit the likelihood they would meet regulatory requirements for importation. We conducted field cage studies to assess the predation capacity and tomato plant damage of three mirid species established in south USA, a known biocontrol agent (Nesidiocoris tenuis), and two native species (Macrolophus praeclarus and Engytatus modestus). All three species significantly reduced the number of whiteflies (Bemisia tabaci) on tomato plants compared to tomato plants without mirids. More damage, evaluated as the number of necrotic rings, was observed on tomato plants with E. modestus and N. tenuis compared to M. praeclarus. In our experiments that included sesame plants (Sesamum indicum) with tomato plants, mirid numbers increased despite a low number of prey, thus showing a benefit of the plant-feeding habit of these predators. USA's established mirids may therefore prove to be immediately available biological agents for the management of present and future tomato pests.

A new synthetic lure for management of the invasive giant African snail, Lissachatina fulica.

Synthetic chemical lures mimicking pheromones or food attractants are essential tools in eradication programs for invasive species. However, their uses in programs aiming to control or eradicate terrestrial gastropods are largely unexplored. The goal of this study was to find a synthetic attractant that could aid in the eradication or management of the giant African snail (Lissachatina fulica). Field studies in Hawaii showed that a commercial papaya-flavored oil attracted snails. Analysis of the odor profile of the oil identified a total of 22 chemicals, which comprised > 98% of the volatile compounds emitted by the oil. A synthetic blend was reconstructed that mirrored the release rates of the papaya oil odors. In laboratory and field bioassays, the reconstructed blend, applied to cotton wicks as water and canola oil or water and mineral emulsions, attracted more snails than the water and oil emulsion control wicks. Field studies in Hawaii and Florida showed that the reconstructed blend in an oil emulsion was not attractive to non-target species such as butterflies or bees. The snails were attracted from distances > 1 m and entered traps baited with the attractant emulsion. When tested in the South Florida giant African snail eradication program, direct ground application of the reconstructed papaya-flavored oil emulsion increased the number of snails killed by over 87% compared to water emulsion controls. Integrating tactics using the synthetic papaya oil attractant into control measures should increase the effectiveness of eradication and management programs.

Designing a trapping strategy to aid Giant African Snail (Lissachatina fulica) eradication programs.

In pest eradication programs, traps can directly reduce pest populations; however, their application to gastropod programs remains relatively unexplored. The South Florida Giant African Snail, Liassachatina fulica (Pulmonata: Achatinidae), eradication program allowed a realistic evaluation of their utility. Field studies were conducted to determine the best bait, barrier and trap for use during the eradication program. Immature and adult snails were attracted to banana fruit and a commercially produced bait but only the commercially produced bait did not attract non-target and pest mammals. Four commercially produced traps and 4 barriers were field evaluated for snail retention efficacy. Snails escaped all traps and trap/barrier combinations but the rate of escape ranged from 10-100% after 24 hrs. Laboratory studies confirmed that snails can survive crossing a 5 cm barrier of copper tape, salt, insect stickem or antifouling paint. In the laboratory study snails did not cross copper sulfate but they crossed the barrier in the field. Adding salt to traps as a means to retain snails reduced the number of snails trapped. Laboratory studies confirmed that dry salt decreased the number of snails entering traps and snails did not enter traps when the salt was dissolved in water. Two trap types and the commercial bait were selected for a large-scale program test. For three months, trapping along with hand collection and pesticide application were conducted on 114 properties in five locations. Traps caught snails when surveys and regular pesticide applications on the same properties did not detect them. On 21 occasions snails were only found in traps, and both immature and adult snails were caught. This study showed that traps could be effectively deployed in an eradication program and they could capture snails that may have escaped other control measures.

Reproductive Ecology of the Giant African Snail in South Florida: Implications for Eradication Programs.

Giant African snail (Achatina fulica (Bowdich, 1822)), an important invasive snail, was recently found in South Florida, USA. An extensive eradication effort was initiated consisting of pesticide applications, debris removal and hand collections. We studied the reproduction capacity and population dynamics of snails collected from 22 populations for two years to help evaluate the likely success of the eradication program. A total of 23,890 snails, ranging from 25-131 mm, were measured, dissected and the number of eggs in each snail counted. Gravid snails ranged from 48-128 mm. Only 5% of snails had eggs, which were found year round. As the snails increased in size, they were more likely to include reproducing individuals. However, the percentage of gravid snails peaked when snails were approximately 90 mm. Although more prevalent, small (<65 mm) adults contributed fewer eggs to the population than the larger snails. We evaluated the effect of control measures on six populations having >1000 adult snails and used data from the two largest populations to investigate how environmental factors (temperature, humidity, and rainfall) interacted with population dynamics and control measures. More snails were collected in weeks with high humidity and more gravid snails were collected when the temperature was higher. The addition of metaldehyde pesticides had the greatest impact on population dynamics by reducing snail numbers. In populations with fewer snails, their numbers were already declining before the use of metaldehyde, although the new treatment accelerated the process. As a consequence of the eradication program, egg-producing snails were no longer collected from most populations by the end of the study. The aggressive and persistent control efforts apparently lead to reduced populations of egg producing snails, eventually resulting in local extinctions of this important pest.

Efficiency of Trapping Systems for Detecting Tuta absoluta (Lepidoptera: Gelechiidae).

Tuta absoluta Meyrick (Lepidoptera: Gelechiidae), a pest of tomato, was recently detected in Panama in Central America and now threatens to expand into the important tomato production areas of Mexico and the United States. Moths caught in T. absoluta pheromone-baited traps must be removed and dissected to confirm the species present before containment and mitigation strategies are put in place. Timely processing of traps can be hindered by the presence of numerous similar nontarget moths that cannot be easily prescreened. Trapping systems using dry bucket traps or Delta traps with either hot melt pressure sensitive adhesives (HMPSA) or cool melt adhesives were evaluated for their effectiveness in trapping T. absoluta and for their ease in allowing identification of nontarget moths. Delta traps in Panama with HMPSA and cool melt adhesives both trapped T. absoluta with equal efficacy. In Florida, nontarget moths were easier to prescreen from bucket traps and HMPSA inserts. Importantly, moths found in bucket traps as well as on cool melt adhesive inserts were of a lower quality than those on HMPSA inserts, making identification more difficult. Studies conducted in Florida and Panama tomato and potato fields showed that commercially produced pheromones containing only the main pheromone component ((3E, 8Z, 11Z)-tetradecatrien-1-yl acetate) or containing both the main and minor pheromone component ((3E, 8Z)-tetradecadien-1-yl) attracted nontarget moths. Survey programs, particularly large-scale ones, should consider the application of alternative trapping systems or new adhesives available in order to facilitate the visual prescreening of nontarget moths.

Host plant range of Raoiella indica (Acari: Tenuipalpidae) in areas of invasion of the New World.

Raoiella indica has spread rapidly through the Neotropical region where the mite damages economically and ecologically important plants. Three studies were conducted to determine the host plant range of R. indica, using the presence of colonies containing all life stages as an indicator of reproductive suitability. Periodic surveys at the Fairchild Tropical Botanic Garden (Miami Dade County, FL, USA) and the Royal Botanical Gardens (Port of Spain, Trinidad and Tobago) identified 27 new reproductive host plants. The reproductive suitability of two dicotyledonous species and three native Florida palm species was examined. An updated list of reproductive host plants of R. indica is presented. All reported reproductive hosts (91 plant species) of R. indica are monocots from the orders Arecales (Arecaceae), Zingiberales (Heliconiaceae, Musaceae, Strelitziaceae, Zingiberaceae) and Pandanales (Pandanaceae). Most are palms of the family Arecaceae that originated in areas of the Eastern Hemisphere; about one fourth of the reported hosts are native to the New World and could be considered new host associations of R. indica. Six years after the initial detection in the Caribbean, R. indica has expanded its host plant range. Here we report 27 new reproductive host of R. indica that represent 30% of increase on previous host plant records. As this mite continues spreading in the Neotropical region a great diversity of plants is potentially affected.

Developing detection and monitoring strategies for Planococcus minor (Hemiptera: Pseudococcidae).

A pheromone-based system to locate and monitor Planococcus minor (Maskell), a pest of over 250 plants including citrus, grape, and cacao, was tested. The difficulty in distinguishing P. minor from the citrus mealybug, P. citri, makes finding and evaluating the impact of the pest challenging. Studies conducted in Puerto Rico determined that synthetic P. minor pheromone lures preaged 120 d in the field caught similar number of males as lures not aged (fresh). Molecular analysis of trapped mealybug males using mitochondrial cytochrome oxidase-1, the internal transcriber space two locus, and 28S-D2 gene showed the pheromone traps to be species specific. Traps baited with P. minor pheromone were used to monitor the pest in south Florida and to locate potential infestations. P. minor males were found at all locations studied in South Florida and were present in low numbers (1.03 +/- 0.69 mean +/- SE/trap/14 d). Over 14,000 terminals, fruit, and flowers were visually inspected over a 6 mo period of peak trap catches before the first adult P. minor female was found. The synthetic pheromone lures proved to be an effective tool to locate and monitor this pest new to the continental United States.

Life history of the mealybug, Maconellicoccus hirsutus (Hemiptera: Pseudococcidae), at constant temperatures.

Important life history parameters of the mealybug, Maconellicoccus hirsutus (Green), were characterized on hibiscus (Hibiscus rosa-sinensis L.) cuttings at six constant temperatures between 15 and 35 degrees C. The development of M. hirsutus was the fastest at 27 degrees C, where the mealybugs completed development in approximately 29 d. The lower (T(min)) and upper (T(max)) developmental thresholds and the optimal developmental temperature (T(opt)) for the development of female mealybugs were estimated as 14.5, 35, and 29 degrees C, respectively. The thermal constant (K), which is the number of temperature-day or degree-day units required for development, of the females was 347 DD. The original distribution range prediction (based on T(min) = 17.5 degrees C and K = 300 DD) indicated that M. hirsutus could complete at least one generation in all of the continental United States. However, results of this study suggested that the distribution range of M. hirsutus may expand northward because of the lower T(min), and the predicted number of generations in a year may be lower because of the higher K required to complete each generation. The average cumulative survival rate of M. hirsutus at 25 and 27 degrees C was 72%, which was significantly higher than 51 and 62% at 20 and 30 degrees C, respectively. M. hirsutus reproduced sexually, with each mated female producing 260-300 eggs between 20 and 27 degrees C but only approximately 100 eggs at 30 degrees C. Female longevity was reduced from 28 d at 20 degrees C to 19-21 d at 25-30 degrees C. At 27 degrees C, the net reproductive rate (R(o)) was estimated at 165 female symbol/female symbol, the intrinsic rate of population increase (r(m)) was 0.119 (female symbol/female symbol/d), the generation time (T(G)) was 43 d, and the doubling time (DT) was 5.8 d. The life table statistics suggested that the currently released biological control agents, which have higher r(m) than M. hirsutus, will be able to complete more generations than the mealybug within the tested temperature range; thus, they are effective against M. hirsutus.

Olfactory discrimination among sex pheromone stereoisomers: chirality recognition by pink hibiscus mealybug males.

Our previous field studies suggested that the two chiral centers in the sex pheromone of pink hibiscus mealybug, Maconellicoccus hirsutus, could elicit different male responses. The chiral center in the acid moiety of the pheromone seemed to be more critical than the alcohol portion of the pheromone molecule for attractiveness. The objective of the current study was to test this hypothesis by deploying stereoisomeric blends in pheromone traps. Captures of male M. hirsutus showed that pheromone with the naturally occurring (R)-maconelliyl (S)-2-methylbutanoate and (R)-lavandulyl (S)-2-methylbutanoate [R-S configuration] was most attractive and that pheromone with the unnatural S-S configuration was less attractive. In addition, the RS-R blend (containing R-R and S-R stereoisomers) yielded captures of male M. hirsutus that were comparable to blank controls, and an inhibitory effect was observed when R-R and S-R were combined with naturally occurring R-S blend. These results suggest a unique chirality recognition mechanism; olfactory discrimination among different pheromone stereoisomers depends upon both asymmetric centers. The S configuration on the acid moiety elicits attraction, whereas the R configuration induces inhibition. However, the attractive activity shows some degree of tolerance toward chirality change in the alcohol portion of the pheromone molecules.

Individual variability in herbivore-specific elicitors from the plant's perspective.

Lepidopteran larvae oral secretions and regurgitant (R), which contain a plethora of potential elicitors, are known to dramatically change a plant's wound response. We demonstrate, with a detailed microarray and secondary metabolite analysis, that the two most abundant fatty acid-amino acid conjugates (FACs) in the R of the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) can account for all measured direct (trypsin proteinase inhibitor: TPI) and indirect (cis-alpha-bergamotene) defences, the endogenous jasmonic acid burst that elicits them, and 86% of the induced transcriptional changes (89% up and 83% down) in its native host Nicotiana attenuata and hence are necessary and sufficient for the Manduca-specific modulation of the wound response. FACs were not found in eggs, but detected in larvae of all instars after their first meal. FACs were found in all regions of the alimentary canal and in the frass, but did not occur in salivary or mandibular glands, extracts of which were not active in any assay. Individual larvae differed substantially in their FAC composition and two FAC chemotypes were discernible: N-linolenoyl-L-glutamine biased R and N-linolenoyl-L-glutamate biased R. We created synthetic blends of FACs to mimic these chemical phenotypes and determined whether plants respond differently to the different R chemotypes. Micorarray and TPI analysis revealed that plants do not differentiate. N. attenuata plants use FACs from feeding caterpillars to tailor their wound responses but do not use the variability in FAC ratios to recognize attack from an individual caterpillar.

Allometric analysis of the induced flavonols on the leaf surface of wild tobacco (Nicotiana attenuata).

Trichomes excrete secondary metabolites that may alter the chemical composition of the leaf surface, reducing damage caused by herbivores, pathogens and abiotic stresses. We examined the surface exudates produced by Nicotiana attenuata Torr. Ex Wats., a plant known to contain and secrete a number of secondary metabolites that are toxic or a deterrent to herbivorous insects. Extractions specific to the leaf surface, the trichomes, and the laminar components demonstrated the localization of particular compounds. Diterpene glycosides occurred exclusively in leaf mesophyll, whereas nicotine was found in both the trichomes and mesophyll. Neither rutin nor nicotine was found on the leaf surface. Quercetin and 7 methylated derivatives were found in the glandular trichomes and appeared to be excreted onto the leaf surface. We examined the elicitation of these flavonols on the leaf surface with a surface-area allometric analysis, which measures changes in metabolites independent of the effects of leaf expansion. The flavonols responded differently to wounding, methyl jasmonate (MeJA), herbivore attack and UV-C radiation, and the response patterns corresponded to their compound-specific allometries. Finding greater amounts of quercetin on younger leaves and reduced amounts after herbivore feeding and MeJA treatment, we hypothesized that quercetin may function as an attractant, helping the insects locate a preferred feeding site. Consistent with this hypothesis, mirids (Tupiocoris notatus) were found more often on mature leaves sprayed with quercetin at a concentration typical of young leaves than on unsupplemented mature leaves. The composition of metabolites on the leaf surface of N. attenuata changes throughout leaf development and in response to herbivore attack or environmental stress, and these changes are mediated in part by responses of the glandular trichomes.