Seasonal Dynamics of Flight Phenology of the Euwallacea fornicatus Species Complex and an Associated Parasitoid Wasp in Avocado Groves in Taiwan.

The Euwallacea fornicatus species complex (Coleoptera: Curculionidae: Scolytinae: Xyleborini) is a group of four cryptic ambrosia beetle species. Native to Asia, several members of the complex have invaded other continents, where they cause significant economic losses to agricultural crops (e.g., avocado) and natural ecosystems. We were primarily interested in developing management strategies by focusing on the flight behavior of the beetles. Thus, seasonal differences in flight activity were assessed using panel traps baited with a commercial quercivorol lure, placed in infested avocado orchards in Danei, Tainan, Taiwan. Same traps were used to investigate the flight activity of a natural enemy, an undescribed species of the Braconid genus Eucosmophorus sp. Shothole borer species were identified using a DNA-based, high resolution melting assay. Trap data were compared to the predictions of a simple degree-day model, incorporating developmental data and several environmental parameters known to influence flight. Such as the time period representing most of flight activity in a day and temperature-dependent flight propensity. In stark contrast to the degree-day model which predicted the highest emergence, and by extension flight, of shothole borers during spring and summer (May to November), flight activity was actually lowest during these months, and instead, peaked during the winter (October to March). Abundance of the parasitoid wasp closely mirrored flight activity of the shothole borers. The mismatch of trapping and modeling data can have many causes, heavy precipitation and possibly cooperative brood care may suppress the dispersal behavior of the shothole borers during the summer.

Tropical bee species abundance differs within a narrow elevational gradient.

Insect pollination is among the most essential ecosystem services for humanity. Globally, bees are the most effective pollinators, and tropical bees are also important for maintaining tropical biodiversity. Despite their invaluable pollination service, basic distributional patterns of tropical bees along elevation gradients are globally scarce. Here, we surveyed bees at 100 m elevation intervals from 800 to 1100 m elevation in Costa Rica to test if bee abundance, community composition and crop visitor assemblages differed by elevation. We found that 18 of 24 bee species spanning three tribes that represented the most abundantly collected bee species showed abundance differences by elevation, even within this narrow elevational gradient. Bee assemblages at the two crop species tested, avocado and squash, showed community dissimilarity between high and low elevations, and elevation was a significant factor in explaining bee community composition along the gradient. Stingless bees (Tribe Meliponini) were important visitors to both crop species, but there was a more diverse assemblage of bees visiting avocado compared to squash. Our findings suggest that successful conservation of tropical montane bee communities and pollination services will require knowledge of which elevations support the highest numbers of each species, rather than species full altitudinal ranges.

Expression of several Phytophthora cinnamomi putative RxLRs provides evidence for virulence roles in avocado.

Phytophthora cinnamomi is a plant pathogenic oomycete that causes Phytophthora root rot of avocado (PRR). Currently, there is a limited understanding of the molecular interactions underlying this disease. Other Phytophthora species employ an arsenal of effector proteins to manipulate host physiology, of which the RxLR effectors contribute to virulence by interfering with host immune responses. The aim of this study was to identify candidate RxLR effectors in P. cinnamomi that play a role in establishing PRR, and to infer possible functions for these effectors. We identified 61 candidate RxLR genes which were expressed during infection of a susceptible avocado rootstock. Several of these genes were present in multiple copies in the P. cinnamomi genome, suggesting that they may contribute to pathogen fitness. Phylogenetic analysis of the manually predicted RxLR protein sequences revealed 12 P. cinnamomi RxLRs that were related to characterised effectors in other Phytophthora spp., providing clues to their functions in planta. Expression profiles of nine more RxLRs point to possible virulence roles in avocado-highlighting a way forward for studies of this interaction. This study represents the first investigation of the expression of P. cinnamomi RxLR genes during the course of avocado infection, and puts forward a pipeline to pinpoint effector genes with potential as virulence determinants, providing a foundation for the future functional characterization of RxLRs that contribute to P. cinnamomi virulence in avocado.

Modifications of the U.S. food and drug administration validated method for detection of Cyclospora cayetanensis oocysts in prepared dishes: Mexican-style salsas and guacamole.

Although multiple outbreak clusters of Cyclospora cayetanensis have been traced back to consumption of dishes in Mexican-style restaurants, the FDA Bacteriological Analytical Manual (BAM) does not currently provide methods to detect C. cayetanensis in dishes that contain multiple produce ingredients, such as salsas and guacamole. These complex food matrices also may contain high levels of fats, which can interfere with the detection. Several modifications to the BAM Chapter 19b method (washing produce, DNA extraction, and a TaqMan real-time PCR assay targeting the 18S rRNA gene of C. cayetanensis) were assessed with the goal to detect as few as 5 oocysts of C. cayetanensis in 25 g samples of commercial salsa/pico de gallo, guacamole, and salsa verde. Both freshly prepared and frozen versions of these foods were seeded with 5, 10 and 200 oocysts. For salsa samples, using a gentler washing step than recommended by BAM, we achieved detection of 5 oocysts in the samples (81.8%, n = 11). Increasing the amount of Alconox® in the wash solution to 1%, rather than the 0.1% used in BAM, and adjusting the DNA extraction protocol to process large wash pellets, enabled detection of 5 oocysts in guacamole. To reach the desired level of detection in salsa verde, two types of modifications were necessary: gentler washing and DNA extraction modifications. The use of these same method modifications on previously frozen food samples, provided levels of detection similar to those achieved with fresh dishes. Our modifications enabled robust and reproducible detection of C. cayetanensis in multi-ingredient Mexican dishes, detecting as few as 5 oocysts in 25 g samples. Validating and deploying effective methods to detect C. cayetanensis in high risk fresh produce and prepared dishes are critically important for prevalence studies and outbreak investigations of this parasite.

Evaluation of semiochemical based push-pull strategy for population suppression of ambrosia beetle vectors of laurel wilt disease in avocado.

Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae and Platypodinae) bore into tree xylem to complete their life cycle, feeding on symbiotic fungi. Ambrosia beetles are a threat to avocado where they have been found to vector a symbiotic fungus, Raffaelea lauricola, the causal agent of the laurel wilt disease. We assessed the repellency of methyl salicylate and verbenone to two putative laurel wilt vectors in avocado, Xyleborus volvulus (Fabricius) and Xyleborus bispinatus (Eichhoff), under laboratory conditions. Then, we tested the same two chemicals released from SPLAT flowable matrix with and without low-dose ethanol dispensers for manipulation of ambrosia beetle populations occurring in commercial avocado. The potential active space of repellents was assessed by quantifying beetle catch on traps placed 'close' (~5-10 cm) and 'far' (~1-1.5 m) away from repellent dispensers. Ambrosia beetles collected on traps associated with all in-field treatments were identified to species to assess beetle diversity and community variation. Xyleborus volvulus was not repelled by methyl salicylate (MeSA) or verbenone in laboratory assays, while X. bispinatus was repelled by MeSA but not verbenone. Ambrosia beetle trap catches were reduced in the field more when plots were treated with verbenone dispensers (SPLAT) co-deployed with low-dose ethanol dispensers than when treated with verbenone alone. Beetle diversity was highest on traps deployed with low-dose ethanol lures. The repellent treatments and ethanol lures significantly altered the species composition of beetles captured in experiment plots. Our results indicate that verbenone co-deployed with ethanol lures holds potential for manipulating ambrosia beetle vectors via push-pull management in avocado. This tactic could discourage immigration and/or population establishment of ambrosia beetles in commercial avocado and function as an additional tool for management programs of laurel wilt.

New Oomycota Fungicides With Activity Against Phytophthora cinnamomi and Their Potential Use for Managing Avocado Root Rot in California.

Phytophthora root rot (PRR), caused by Phytophthora cinnamomi, is the most destructive disease of avocado worldwide. In the United States, mefenoxam and phosphonate products are currently the only registered fungicides for managing avocado PRR. Four new Oomycota-specific and two registered fungicides, all with different modes of action, were evaluated. Seventy-one isolates of P. cinnamomi from avocado in California, most of them collected between 2009 to 2017, were tested for their in vitro sensitivity to the six fungicides. Baseline sensitivity ranges and mean values (in parentheses) of effective concentrations to inhibit mycelial growth by 50% (EC50) for the new fungicides ethaboxam, fluopicolide, mandipropamid, and oxathiapiprolin were 0.017 to 0.069 µg/ml (0.035), 0.046 to 0.330 µg/ml (0.133), 0.003 to 0.011 µg/ml (0.005), and 0.0002 to 0.0007 µg/ml (0.0004), respectively. In comparison, the EC50 value range (mean) was 0.023 to 0.138 µg/ml (0.061) for mefenoxam and 12.9 to 361.2 µg/ml (81.5) for potassium phosphite. Greenhouse soil inoculation trials with 8-month-old Zutano seedlings and 10-month-old Dusa and PS.54 clonal rootstocks were conducted to assess the efficacy of these fungicides for managing PRR. Mefenoxam and potassium phosphite were effective treatments; however, oxathiapiprolin, fluopicolide, and mandipropamid were more effective. Ethaboxam was effective in reducing PRR on the rootstocks evaluated. Oxathiapiprolin reduced PRR incidence and pathogen population size in the soil by >90%, and plant shoot growth and root dry weight were significantly increased compared with the control; thus, oxathiapiprolin was one of the best treatments overall. The high activity and performance of these new fungicides supports their registrations on avocado for use in rotation and mixture programs, including with previously registered compounds, to reduce the risk of development and spread of resistance in pathogen populations.

Survey in Nurseries and Irrigation Water Reservoirs as Sources of Oomycetes Found in Avocado Orchards in the Canary Islands.

Nursery stocks and irrigation water are important sources of Phytophthora spp. In this study, 20 irrigation water reservoirs and 10 avocado nurseries were surveyed in the Canary Islands between 2013 and 2015 to investigate their potential role in disseminating pathogenic species in avocado orchards. Phytophthora multivora was isolated from one of the irrigation reservoirs, whereas, in two surveys conducted in nurseries, Phytophthora cinnamomi, the primary pathogen in avocado, was detected in addition to other Phytophthora species (P. lacustris, P. multivora, P. nicotianae, P. niederhauserii, and P. palmivora) and Phytopythium vexans. The species recovered from nurseries were isolated mostly from propagated plants but also from nursery irrigation water, soil used for substrate preparation, and soil samples collected in orchards that supply seeds for seedling propagation. Species recovered from nurseries correlated with those found in avocado orchards in a previous study, except for P. lacustris, suggesting that nurseries could be involved in their dissemination in avocado orchards. The improved sanitary status of nurseries resulted in reduced incidence in the second survey, indicating the importance of nursery monitoring to reduce infestations.

New microsatellite markers for population studies of Phytophthora cinnamomi, an important global pathogen.

Phytophthora cinnamomi is the causal agent of root rot, canker and dieback of thousands of plant species around the globe. This oomycete not only causes severe economic losses but also threatens natural ecosystems. In South Africa, P. cinnamomi affects eucalyptus, avocado, macadamia and indigenous fynbos. Despite being one of the most important plant pathogens with a global distribution, little information is available regarding origin, invasion history and population biology. This is partly due to the limited number of molecular markers available for studying P. cinnamomi. Using available genome sequences for three isolates of P. cinnamomi, sixteen polymorphic microsatellite markers were developed as a set of multiplexable markers for both PCR and Gene Scan assays. The application of these markers on P. cinnamomi populations from avocado production areas in South Africa revealed that they were all polymorphic in these populations. The markers developed in this study represent a valuable resource for studying the population biology and movement of P. cinnamomi and will aid in the understanding of the origin and invasion history of this important species.

Epidemiology, pathology and identification of Colletotrichum including a novel species associated with avocado (Persea americana) anthracnose in Israel.

Anthracnose disease caused by Colletotrichum species is a major constraint for the shelf-life and marketability of avocado fruits. To date, only C. gloeosporioides sensu lato and C. aenigma have been reported as pathogens affecting avocado in Israel. This study was conducted to identify and characterize Colletotrichum species associated with avocado anthracnose and to determine their survival on different host-structures in Israel. The pathogen survived and over-wintered mainly on fresh and dry leaves, as well as fresh twigs in the orchard. A collection of 538 Colletotrichum isolates used in this study was initially characterized based on morphology and banding patterns generated according to arbitrarily primed PCR to assess the genetic diversity of the fungal populations. Thereafter, based on multi-locus phylogenetic analyses involving combinations of ITS, act, ApMat, cal, chs1, gapdh, gs, his3, tub2 gene/markers; eight previously described species (C. aenigma, C. alienum, C. fructicola, C. gloeosporioides sensu stricto, C. karstii, C. nupharicola, C. siamense, C. theobromicola) and a novel species (C. perseae) were identified, as avocado anthracnose pathogens in Israel; and reconfirmed after pathogenicity assays. Colletotrichum perseae sp. nov. and teleomorph of C. aenigma are described along with comprehensive morphological descriptions and illustrations, for the first time in this study.

α-Copaene is an attractant, synergistic with quercivorol, for improved detection of Euwallacea nr. fornicatus (Coleoptera: Curculionidae: Scolytinae).

The tea shot-hole borer, Euwallacea fornicatus Eichhoff, is an ambrosia beetle endemic to Asia and a pest of commercial tea, Camellia sinensis (L.) Kuntze. Recently, a complex of species morphologically similar to E. fornicatus has been recognized, which includes new pests established in Israel and the USA, both in California and Florida. Collectively termed E. nr. fornicatus, these cryptic species carry symbiotic Fusarium spp. fungi, some of which cause dieback disease in susceptible hosts, which include avocado, Persea americana Miller. Due to the threat to this economically important crop, research was initiated to evaluate efficacy of kairomone-based lures for detection of the beetle in Florida (termed the Florida tea shot hole borer, FL-TSHB). A series of field tests were conducted in 2016 in commercial avocado groves known to have FL-TSHB at various population levels. All tests evaluated lures containing quercivorol (p-menth-2-en-1-ol) and α-copaene, presented separately and in combination; and one test evaluated effect of trap type on beetle captures. In addition, electroantennography (EAG) was used to quantify female olfactory responses to lure emissions. This study identified (-)-α-copaene as a new attractant for FL-TSHB, equivalent in efficacy to quercivorol (the standard lure for Euwallacea detection in the USA); however, the combination of lures captured significantly more FL-TSHB than either lure alone. This combination resulted in synergistic attraction at two field sites and additive attraction at a third site. Sticky panel traps captured more FL-TSHB than comparably-baited Lindgren funnel traps. Females engaged in host-seeking flight from 11:00 to 16:00 hr (EST), with peak numbers observed between 12:00 and 13:00 hr. EAG analyses confirmed olfactory chemoreception of both kairomones, with a higher response elicited with the combination of volatiles. Results indicate that detection of pest E. nr. fornicatus in Florida can be improved by using a two-component lure consisting of p-menth-2-en-1-ol and (-)-α-copaene.

Global Potential Distribution of Bactrocera carambolae and the Risks for Fruit Production in Brazil.

The carambola fruit fly, Bactrocera carambolae, is a tephritid native to Asia that has invaded South America through small-scale trade of fruits from Indonesia. The economic losses associated with biological invasions of other fruit flies around the world and the polyphagous behaviour of B. carambolae have prompted much concern among government agencies and farmers with the potential spread of this pest. Here, ecological niche models were employed to identify suitable environments available to B. carambolae in a global scale and assess the extent of the fruit acreage that may be at risk of attack in Brazil. Overall, 30 MaxEnt models built with different combinations of environmental predictors and settings were evaluated for predicting the potential distribution of the carambola fruit fly. The best model was selected based on threshold-independent and threshold-dependent metrics. Climatically suitable areas were identified in tropical and subtropical regions of Central and South America, Sub-Saharan Africa, west and east coast of India and northern Australia. The suitability map of B. carambola was intersected against maps of fruit acreage in Brazil. The acreage under potential risk of attack varied widely among fruit species, which is expected because the production areas are concentrated in different regions of the country. The production of cashew is the one that is at higher risk, with almost 90% of its acreage within the suitable range of B. carambolae, followed by papaya (78%), tangerine (51%), guava (38%), lemon (30%), orange (29%), mango (24%) and avocado (20%). This study provides an important contribution to the knowledge of the ecology of B. carambolae, and the information generated here can be used by government agencies as a decision-making tool to prevent the carambola fruit fly spread across the world.

Cryptophlebia Walsingham, 1900, Thaumatotibia Zacher, 1915, and Archiphlebia Komai & Horak, 2006, in Australia (Lepidoptera: Tortricidae: Olethreutinae: Grapholitini).

The Australian species of the grapholitine genera Cryptophlebia Walsingham, 1900, Thaumatotibia Zacher, 1915 and Archiphlebia Komai & Horak, 2006, are revised, described and illustrated. A key to species is provided. Five named species of Cryptophlebia, C. ombrodelta (Lower), C. iridosoma (Meyrick), C. rhynchias (Meyrick) and C. pallifimbriana Bradley, are redescribed and three new species, C. wraggae, sp. nov., C. stigmata, sp. nov., and C. caulicola, sp. nov., are described. Cryptophlebia amblyopa Clarke, described from Micronesia, is synonymised with C. iridosoma. Cryptophlebia caulicola, sp. nov., is a borer in twigs of Acacia mangium Willd. in northern Queensland. Thaumatotibia aclyta (Turner) and T. zophophanes (Turner) are redescribed and the new species T. maculata, sp. nov., is described. Fruit of Acronychia spp. (Rutaceae) have been identified as native hosts of T. zophophanes, a pest species which damages macadamia (Proteaceae) and avocado (Lauraceae) on the Atherton Tableland. Archiphlebia endophaga (Meyrick) and A. rutilescens (Turner) are redescribed, and the new species A. gilva, sp. nov., is described.

The Scirtothrips perseae species-group (Thysanoptera), with one new species from avocado, Persea americana.

Following recent molecular studies on avocado thrips, a new species is described from Costa Rica, Ecuador, and Colombia from the young leaves of avocado, Persea americana. Scirtothrips hansoni sp.n. is closely related to the Californian pest, S. perseae, and also to S. astrictus from Costa Rica that remains known from a single female. An illustrated key to these three species is provided.

Sampling method evaluation and empirical model fitting for count data to estimate densities of Oligonychus perseae (Acari: Tetranychidae) on 'Hass' avocado leaves in southern California.

Oligonychus perseae (Acari: Tetranychidae) is an important foliar spider mite pest of 'Hass' avocados in several commercial production areas of the world. In California (USA), O. perseae densities in orchards can exceed more than 100 mites per leaf and this makes enumerative counting prohibitive for field sampling. In this study, partial enumerative mite counts along half a vein on an avocado leaf, an industry recommended practice known as the "half-vein method", was evaluated for accuracy using four data sets with a combined total of more than 485,913 motile O. perseae counted on 3849 leaves. Sampling simulations indicated that the half-vein method underestimated mite densities in a range of 15-60 %. This problem may adversely affect management of this pest in orchards and potentially compromise the results of field research requiring accurate mite density estimation. To address this limitation, four negative binomial regression models were fit to count data in an attempt to rescue the half-vein method for estimating mite densities. These models were incorporated into sampling plans and evaluated for their ability to estimate mite densities on whole leaves within 30-tree blocks of avocados. Model 3, a revised version of the original half-vein model, showed improvement in providing reliable estimates of O. perseae densities for making assessments of general leaf infestation densities across orchards in southern California. The implications of these results for customizing the revised half-vein method as a potential field sampling tool and for experimental research in avocado production in California are discussed.

Attraction of Redbay Ambrosia Beetle, Xyleborus Glabratus, To Leaf Volatiles of its Host Plants in North America.

The redbay ambrosia beetle, Xyleborus glabratus, is an important pest of redbay (Persea borbonia) and swamp bay (P. palustris) trees in forests of the southeastern USA. It is also a threat to commercially grown avocado. The beetle is attracted to host wood volatiles, particularly sesquiterpenes. Contrary to other ambrosia beetles that attack stressed, possibly pathogen-infected, and dying trees, X. glabratus readily attacks healthy trees. To date little is known about the role of leaf volatiles in the host selection behavior and ecology of X. glabratus. To address this question, an olfactometer bioassay was developed to test the behavioral response of X. glabratus to plant leaf volatiles. We found that X. glabratus was attracted to the leaf odors of their hosts, redbay and swamp bay, with no attraction to a non-host tree tested (live oak, Quercus virginiana), which served as a negative control. Gas chromatography-mass spectrometry (GS/MS) analysis of leaves revealed the absence of sesquiterpenes known to be attractive to X. glabratus and present in host wood, suggesting that additional leaf-derived semiochemicals may serve as attractants for this beetle. An artificial blend of chemicals was developed based on GC/MS analyses of leaf volatiles and behavioral assays. This blend was attractive to X. glabratus at a level that rivaled currently used lures for practical monitoring of this pest. This synthetic redbay leaf blend also was tested in the field. Baited traps captured more X. glabratus than unbaited controls and equivalently to manuka oil lures. We hypothesize that leaf volatiles may be used by X. glabratus as an additional cue for host location.

Fusarium euwallaceae sp. nov.--a symbiotic fungus of Euwallacea sp., an invasive ambrosia beetle in Israel and California.

The invasive Asian ambrosia beetle Euwallacea sp. (Coleoptera, Scolytinae, Xyleborini) and a novel Fusarium sp. that it farms in its galleries as a source of nutrition causes serious damage to more than 20 species of live trees and pose a serious threat to avocado production (Persea americana) in Israel and California. Adult female beetles are equipped with mandibular mycangia in which its fungal symbiont is transported within and from the natal galleries. Damage caused to the xylem is associated with disease symptoms that include sugar or gum exudates, dieback, wilt and ultimately host tree mortality. In 2012 the beetle was recorded on more than 200 and 20 different urban landscape species in southern California and Israel respectively. Euwallacea sp. and its symbiont are closely related to the tea shot-hole borer (E. fornicatus) and its obligate symbiont, F. ambrosium occurring in Sri Lanka and India. To distinguish these beetles, hereafter the unnamed xyleborine in Israel and California will be referred to as Euwallacea sp. IS/CA. Both fusaria exhibit distinctive ecologies and produce clavate macroconidia, which we think might represent an adaption to the species-specific beetle partner. Both fusaria comprise a genealogically exclusive lineage within Clade 3 of the Fusarium solani species complex (FSSC) that can be differentiated with arbitrarily primed PCR. Currently these fusaria can be distinguished only phenotypically by the abundant production of blue to brownish macroconidia in the symbiont of Euwallacea sp. IS/CA and their rarity or absence in F. ambrosium. We speculate that obligate symbiosis of Euwallacea and Fusarium, might have driven ecological speciation in these mutualists. Thus, the purpose of this paper is to describe and illustrate the novel, economically destructive avocado pathogen as Fusarium euwallaceae sp. nov. S. Freeman et al.

An inordinate fondness for Fusarium: phylogenetic diversity of fusaria cultivated by ambrosia beetles in the genus Euwallacea on avocado and other plant hosts.

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naïve natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Clade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene ∼21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization.

Export of commercial Hass avocados from Argentina poses negligible risk of Ceratitis capitata (Diptera: Tephritidae) infestation.

Argentina has to meet quarantine restrictions because of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), to export 'Hass' avocados, Persea americana Miller, to certain countries. Hass avocado at the hard, mature green stage is potentially a conditional nonhost for C. capitata and could open export markets without the need for a quarantine treatment. Trapping data from 1998 to 2006 showed that C. capitata was present in avocado orchards, particularly early in the harvest season. The host status of hard, mature green Hass avocado to C. capitata was evaluated using laboratory and field cage tests under no-choice conditions and by assessing natural levels of infestation in commercially harvested fruit from the main avocado production area. In total, 2,250 hard, mature green avocado fruit were exposed to 11,250 gravid females for 24 or 48 h after harvest in laboratory or field cages, and no infestations were found. During 11 seasons, 5,949 fruit in total were sampled from the trees and 992 fruit were collected from the ground, and in none of them were any live or dead fruit fly larvae found. Inspection of >198,000 commercial fruit at the packinghouse from 1998 to 2011 showed no symptoms of fruit fly infestation. These data exceed the published standards for determination of nonhost status, as well as the Probit 9 standard for development of quarantine treatments. Hass avocado harvested at the hard, mature green stage was not infested by C. capitata and seems to pose a negligible quarantine risk. As a consequence, no postharvest treatment or other quarantine actions should be required by importing countries.

Sequential hypothesis testing with spatially correlated presence-absence data.

A pest management decision to initiate a control treatment depends upon an accurate estimate of mean pest density. Presence-absence sampling plans significantly reduce sampling efforts to make treatment decisions by using the proportion of infested leaves to estimate mean pest density in lieu of counting individual pests. The use of sequential hypothesis testing procedures can significantly reduce the number of samples required to make a treatment decision. Here we construct a mean-proportion relationship for Oligonychus perseae Tuttle, Baker, and Abatiello, a mite pest of avocados, from empirical data, and develop a sequential presence-absence sampling plan using Bartlett's sequential test procedure. Bartlett's test can accommodate pest population models that contain nuisance parameters that are not of primary interest. However, it requires that population measurements be independent, which may not be realistic because of spatial correlation of pest densities across trees within an orchard. We propose to mitigate the effect of spatial correlation in a sequential sampling procedure by using a tree-selection rule (i.e., maximin) that sequentially selects each newly sampled tree to be maximally spaced from all other previously sampled trees. Our proposed presence-absence sampling methodology applies Bartlett's test to a hypothesis test developed using an empirical mean-proportion relationship coupled with a spatial, statistical model of pest populations, with spatial correlation mitigated via the aforementioned tree-selection rule. We demonstrate the effectiveness of our proposed methodology over a range of parameter estimates appropriate for densities of O. perseae that would be observed in avocado orchards in California.

Evaluation of neonicotinoid, organophosphate and avermectin trunk injections for the management of avocado thrips in California avocado groves.

BACKGROUND: Trunk injections of systemic insecticides were evaluated for the management of avocado thrips. Insecticide residues were quantified in leaves to determine when after treatment, and for how long, toxic concentrations of the insecticides were present. Residues in fruit were quantified to determine whether trunk injection of insecticides might present a greater risk than traditional application methods for contaminating fruit. RESULTS: Residues of imidacloprid and dinotefuran were at least tenfold higher in leaves when trees were treated via trunk injection compared with soil application. Dinotefuran uptake was more rapid than imidacloprid, and no residues were detected within fruit. Acephate was also mobilized very rapidly and gave good control of thrips in bioassays; however, residues of acephate and its insecticidal metabolite methamidophos were detected in the fruit for up to 4 weeks after injection. Avermectin uptake was very slow, and it was ineffective against avocado thrips. CONCLUSIONS: Trunk injections of acephate and dinotefuran permitted rapid uptake into avocados, and they are strong candidates as control methods for avocado thrips. However, residues of organophosphates in fruit could necessitate increased preharvest intervals. Residues of neonicotinoids were below detection limits in fruit, suggesting that neonicotinoids may be the more suitable control option of the two chemical classes.