Risk Assessment of Ozone Fumigation Under Vacuum to Control Potential Infestation of Coffee Berry Borer and Coffee Leaf Rust in Green Coffee Beans Imported Into Hawaii.

Studies were conducted with ozone gas fumigation under vacuum as a methyl bromide alternative against life stages of coffee berry borer (CBB) Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), and the urediniospores of coffee leaf rust (CLR), Hemileia vastatrix Berkeley & Broome (Basidiomycota: Pucciniales) in green coffee, Coffea spp. L. Fumigation with 10,000 ppm O3 gas under -25.4 mm Hg vacuum1 at 13.0 ± 3.0°C for 6.0 h killed all CBB larvae, pupae, and adults, but did not kill all CBB eggs (~15% survival). Mortality of CLR urediniospores was 100% within the first hour of the 6-h fumigation. Ozone fumigation had no adverse effects on coffee quality. Results indicated that CBB adult hitchhikers may be the only target life stage of quarantine concern, and additional studies focused on this stage. CBB adult survival and reproduction decreased significantly at moisture contents ≤20%, and F1 generation survival did not occur in green coffee at moisture contents ≤15%. As the international standard for green coffee moisture content is 9-12%, adult CBB should not survive or reproduce in exported dry green coffee. Standard industry processing of harvested coffee cherries to the green coffee stage using either mechanical- or sun-drying eliminated CBB infestations from the field. A systems approach is recommended for exporting green coffee to control CBB and CLR that includes eliminating CBB life stages with standard processing methods, reducing moisture content to 9-12% to prevent egg deposition, survival or reproduction, and O3 fumigation to ensure quarantine security against potential CBB adult hitchhikers.

Evolutionary history of the little fire ant Wasmannia auropunctata before global invasion: inferring dispersal patterns, niche requirements and past and present distribution within its native range.

The evolutionary history of invasive species within their native range may involve key processes that allow them to colonize new habitats. Therefore, phylogeographic studies of invasive species within their native ranges are useful to understand invasion biology in an evolutionary context. Here we integrated classical and Bayesian phylogeographic methods using mitochondrial and nuclear DNA markers with a palaeodistribution modelling approach, to infer the phylogeographic history of the invasive ant Wasmannia auropunctata across its native distribution in South America. We discuss our results in the context of the recent establishment of this mostly tropical species in the Mediterranean region. Our Bayesian phylogeographic analysis suggests that the common ancestor of the two main clades of W. auropunctata occurred in central Brazil during the Pliocene. Clade A would have differentiated northward and clade B southward, followed by a secondary contact beginning about 380,000 years ago in central South America. There were differences in the most suitable habitats among clades when considering three distinct climatic periods, suggesting that genetic differentiation was accompanied by changes in niche requirements, clade A being a tropical lineage and clade B a subtropical and temperate lineage. Only clade B reached more southern latitudes, with a colder climate than that of northern South America. This is concordant with the adaptation of this originally tropical ant species to temperate climates prior to its successful establishment in the Mediterranean region. This study highlights the usefulness of exploring the evolutionary history of invasive species within their native ranges to better understand biological invasions.

Large-scale confirmatory tests of a phytosanitary irradiation treatment against Sternochetus frigidus (Coleoptera: Curculionidae) in Philippine mango.

The mango pulp weevil, Sternochetus frigidus (F.), is an important quarantine pest preventing the export of mangoes from the Philippines to the United States and other countries. Previously, a radiation dose of 100 Gy was proposed for phytosanitary treatment of S. frigidus based on dose-response studies with larvae, pupae, and adult weevils. To validate an irradiation treatment, large-scale confirmatory tests were conducted with adults (the most radiation-tolerant stage) in mangoes at 100 and 150 Gy. After treatment, adults were removed from fruit, sexed, and mated in pairs to observe any reproduction. At 100 Gy, adults laid a small number of eggs but none of the eggs hatched. At 150 Gy (measured doses 96.7-164.1 Gy),4,559 treated weevils laid no eggs, indicating that this dose caused complete sterility. Irradiation treatment with a minimum absorbed dose of 165 Gy will therefore provide quarantine security for S. frigidus in exported Philippine mangoes.

Toward multi-scale computational modeling in developmental disability research.

The field of theoretical neuroscience is gaining increasing recognition. Virtually all areas of neuroscience offer potential linkage points for computational work. In developmental neuroscience, main areas of research are neural development and connectivity, and connectionist modeling of cognitive development. In this paper, we suggest that computational models can be helpful tools for understanding the pathogenesis and consequences of perinatal brain damage and subsequent developmental disability. In particular, designing multi-scale computational models should be considered by developmental neuroscientists interested in helping reduce the risk for developmental disabilities.

Long-term patterns and feeding sites of southern green stink bug (Hemiptera: Pentatomidae) in Hawaii macadamia orchards, and sampling for management decisions.

Southern green stink bug (Nezara viridula, Hemiptera: Pentatomidae) is a pest of macadamia nuts, causing pitting to kernels by feeding. In spite of its pest status, many aspects of the ecology of this insect in macadamia orchards are poorly understood. This study analyzes long-term N. viridula damage to macadamia nuts and investigates the extent to which damage to nuts occurs in the tree canopy, prior to nut-drop. We show that there are distinct seasonal peaks in damage detected after harvest and that, over six years of data collection, mean damage levels were fairly low, albeit with spikes in damage levels recorded. Sampling nuts at peak harvest periods from different strata in the trees and from the ground showed that incidence of damaged nuts within the canopy was typically half as high as on the fallen nuts. Damage to fallen nuts may have occurred prior to nut-drop, and continued to accumulate after nut-drop. These results show that management of N. viridula within macadamia canopies, as opposed to only on fallen nuts, is important. A sampling procedure and predictive model for estimating late-season damage based on early-season damage samples is provided. The model uses January and March damage measurements (based on samples with set level of accuracy), mean temperature and month of the year for which damage is predicted. Early-season damage of 6-10% predicts late-season damage levels that should justify N. viridula suppression based on the nominal threshold (13% damage) used by kernel processors to reject nuts based on damage.

Nontarget effects--the Achilles' heel of biological control? Retrospective analyses to reduce risk associated with biocontrol introductions.

Controversy exists over ecological risks in classical biological control. We reviewed 10 projects with quantitative data on nontarget effects. Ten patterns emerged: (a) Relatives of the pest are most likely to be attacked; (b) host-specificity testing defines physiological host range, but not ecological range; (c) prediction of ecological consequences requires population data; (d) level of impact varied, often in relation to environmental conditions; (e) information on magnitude of nontarget impact is sparse; (f) attack on rare native species can accelerate their decline; (g) nontarget effects can be indirect; (h) agents disperse from agroecosystems; (i) whole assemblages of species can be perturbed; and (j) no evidence on adaptation is available in these cases. The review leads to six recommendations: Avoid using generalists or adventive species; expand host-specificity testing; incorporate more ecological information; consider ecological risk in target selection; prioritize agents; and pursue genetic data on adaptation. We conclude that retrospective analyses suggest clear ways to further increase future safety of biocontrol.

First Report of Lasmenia sp. and Two Species of Gliocephalotrichum on Rambutan in Hawaii.

Rambutan (Nephelium lappaceum L.) is a tropical fruit grown in Hawaii for the exotic fruit market. Fruit rot was observed periodically during 1998 and 1999 from two islands, Hawaii and Kauai, and severe fruit rot was observed during 2000 in orchards in Kurtistown and Papaikou on Hawaii. Symptoms were characterized by brown-to-black, water-soaked lesions on the fruit surface that progressed to blackening and drying of the pericarp, which often split and exposed the aril (flesh). In certain cultivars, immature, small green fruits were totally mummified. Rambutan trees with high incidence of fruit rot also showed symptoms of branch dieback and leaf spot. Lasmenia sp. Speg. sensu Sutton, identified by Centraalbureau voor Schimmelcultures (Baarn, the Netherlands), was isolated from infected fruit and necrotic leaves. Also associated with some of the fruit rot and dieback symptoms were Gliocephalotrichum simplex (J.A. Meyer) B. Wiley & E. Simmons, and G. bulbilium J.J. Ellis & Hesseltine. G. simplex was isolated from infected fruit, and G. bulbilium was isolated from discolored vascular tissues and infected fruit. Identification of species of Gliocephalotrichum was based on characteristics of conidiophores, sterile hairs, and chlamydospores (1,4). Culture characteristics were distinctive on potato dextrose agar (PDA), where the mycelium of G. bulbilium was light orange (peach) without reverse color, while G. simplex was golden-brown to grayish-yellow with dark brown reverse color. Both species produced a fruity odor after 6 days on PDA. In pathogenicity tests, healthy, washed rambutan fruits were wounded, inoculated with 30 µl of sterile distilled water (SDW) or a fungus spore suspension (105 to 106 spores per ml), and incubated in humidity chambers at room temperature (22°C) under continuous fluorescent light. Lasmenia sp. (strain KN-F99-1), G. simplex (strain KN-F2000-1), and G. bulbilium (strains KN-F2001-1 and KN-F2001-2) produced fruit rot symptoms on inoculated fruit and were reisolated from fruit with typical symptoms, fulfilling Koch's postulates. Controls (inoculated with SDW) had lower incidence or developed less severe symptoms than the fungus treatments. Inoculation tests were conducted at least twice. To our knowledge, this is the first report of Lasmenia sp. in Hawaii and the first report of the genus Gliocephalotrichum on rambutan in Hawaii. These pathogens are potentially economically important to rambutan in Hawaii. G. bulbilium has been reported previously on decaying wood of guava (Psidium guajava L.) in Hawaii (2), and the fungus causes field and postharvest rots of rambutan fruit in Thailand (3). References: (1) J. J. Ellis and C. W. Hesseltine. Bull. Torrey Bot. Club 89:21, 1962. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (3) N. Visarathanonth and L. L. Ilag. Pages 51-57 in: Rambutan: Fruit Development, Postharvest Physiology and Marketing in ASEAN. ASEAN Food Handling Bureau, Kuala Lumpur, Malaysia, 1987. (4) B. J. Wiley and E. G. Simmons. Mycologia 63:575, 1971.

Accelerated development of quarantine treatments for insects on poor hosts.

The probit 9 standard for quarantine treatment efficacy (99.9968% mortality) was originally recommended for tropical fruits heavily infested with fruit flies and it centers on high mortality to achieve quarantine security. This standard may be too stringent for quarantine pests in commodities that are rarely infested or are poor hosts, The alternative treatment efficacy approach measures risk as the probability of a mating pair, gravid female, or parthenogenic individual surviving in a shipment. This will be a function of many factors including infestation rate and shipment volume. Applying the risk-based alternative treatment efficacy approach to pests on rarely infested or poor hosts will lower the number of required test insects needed for developing quarantine treatments; hence data for a quarantine treatment could be generated by testing 10,000 or fewer insects with no survivors, compared with 90,000-100,000 insects to demonstrate the traditional probit 9 efficacy. Several commodity/quarantine pest systems where this approach could be applied are discussed. This approach would save time and resources, and help farmers export their crop on a more-timely basis.

Hot water immersion to ensure quarantine security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in lychee and longan exported from Hawaii.

We determined whether immersion in 49 degrees C water for 20 min, a quarantine treatment developed for disinfestation of fruit flies in lychee, Litchi chinensis Sonn., and longan, Dimocarpus longan (Lourd.) Steud., exported from Hawaii, would also disinfest fruit of two species of Cryptophlebia. The pattern of tolerance to heat in Cryptophlebia illepida (Butler) was generally eggs < neonates < early instars = late instars < pupae. No C. illepida survived immersion for 16 or 20 min. Late fourth and fifth instars were determined to be the most tolerant stage that occurs in harvested fruit. Late instars of Cryptophlebia ombrodelta (Lower) were more tolerant of hot-water immersion than those of C. illepida, but no C. ombrodelta late instars survived immersion for 16 or 20 min. The hot water immersion quarantine treatment for fruit flies should effectively disinfest lychees and longans of any Cryptophlebia.

Effect of mango weevil (Coleoptera: Curculionidae) damage on mango seed viability in Hawaii.

The mango weevil, Cryptorhynchus (= Sternochetus) mangiferae (F.), is a federally quarantined pest that prevents shipment of mangos from Hawaii into the continental United States. Although this monophagous weevil allegedly causes reduced seed germination, damage to the fruit pulp, and premature fruit drop in mangos, there are few studies examining these potential sources of crop loss. We conducted studies to assess the effect of mango weevil infestation on seed viability while making observations on the frequency of pulp feeding. Naturally infested seeds from mature fruit were planted in pots and scored for successful germination. Germination rates for infested seeds were equal to those of uninfested control seeds in a polyembryonic cultivar ('Common'), whereas germination was significantly reduced for infested seeds of a monoembryonic cultivar ('Haden') compared with uninfested control seeds but germination of infested seeds was still > 70%. To assess seed tolerance of damage, seeds were artificially damaged by cutting away 25, 50, or 75% of the cotyledon before planting and scored for germination. None of the damage treatments was significantly different from the undamaged controls, indicating that mango seeds can withstand substantial damage and still germinate successfully. Over the 2-yr period we conducted experiments, only four of 3,602 mango fruits (0.11%) showed evidence of direct feeding damage to the pulp. Results suggest that C. mangiferae is a less serious pest of mangos than previously thought.

Field infestation of rambutan fruits by internal-feeding pests in Hawaii.

More than 47,000 mature fruits of nine different varieties of rambutan (Nephelium lappaceum L.) were harvested from orchards in Hawaii to assess natural levels of infestation by tephritid fruit flies and other internal feeding pests. Additionally, harvested, mature fruits of seven different rambutan varieties were artificially infested with eggs or first-instars of Mediterranean fruit fly, Ceratitis capitata (Wiedemann), or oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) to assess host suitability. When all varieties were combined over two field seasons of sampling, fruit infestation rates were 0.021% for oriental fruit fly, 0.097% for Cryptophlebia spp. (Lepidoptera: Tortricidae), and 0.85% for pyralids (Lepidoptera). Species of Cryptophlebia included both C. illepida (Butler), the native Hawaiian species, and C. ombrodelta (Lower), an introduced species from Australia. Cryptophlebia spp. had not previously been known to attack rambutan. The pyralid infestation was mainly attributable to Cryptoblabes gnidiella (Milliere), a species also not previously recorded on rambutan in Hawaii. Overall infestation rate for other moths in the families Blastobasidae, Gracillariidae, Tineidae, and Tortricidae was 0.061%. In artificially infested fruits, both species of fruit fly showed moderately high survivorship for all varieties tested. Because rambutan has such low rates of infestation by oriental fruit fly and Cryptophlebia spp., the two primary internal-feeding regulatory pests of rambutan in Hawaii, it may be amenable to the alternative treatment efficacy approach to postharvest quarantine treatment.

NBQX attenuates excitotoxic injury in developing white matter.

The excitatory neurotransmitter glutamate is released from axons and glia under hypoxic/ischemic conditions. In vitro, oligodendrocytes (OLs) express non-NMDA glutamate receptors (GluRs) and are susceptible to GluR-mediated excitotoxicity. We evaluated the role of GluR-mediated OL excitotoxicity in hypoxic/ischemic white matter injury in the developing brain. Hypoxic/ischemic white matter injury is thought to mediate periventricular leukomalacia, an age-dependent white matter lesion seen in preterm infants and a common antecedent to cerebral palsy. Hypoxia/ischemia in rat pups at postnatal day 7 (P7) produced selective white matter lesions and OL death. Furthermore, OLs in pericallosal white matter express non-NMDA GluRs at P7. Unilateral carotid ligation in combination with hypoxia (6% O(2) for 1 hr) resulted in selective, subcortical white matter injury with a marked ipsilateral decrease in immature and myelin basic protein-expressing OLs that was also significantly attenuated by 6-nitro-7-sulfamoylbenzo(f)quinoxaline-2,3-dione (NBQX). Intracerebral AMPA demonstrated greater susceptibility to OL injury at P7 than in younger or older pups, and this was attenuated by systemic pretreatment with the AMPA antagonist NBQX. These results indicate a parallel, maturation-dependent susceptibility of immature OLs to AMPA and hypoxia/ischemia. The protective efficacy of NBQX suggests a role for glutamate receptor-mediated excitotoxic OL injury in immature white matter in vivo.

Irradiation to ensure quarantine security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in sapindaceous fruits from Hawaii.

Studies were undertaken to determine whether irradiation treatment at 250 Gy, an accepted treatment for disinfestation of fruit flies in spindaceous fruits from Hawaii, would also disinfest fruit of two species of Cryptophlebia. Cryptophlebia illepida (Butler) was determined to be more tolerant of irradiation than Cryptophlebia ombrodelta (Lower); therefore, C. illepida was the focus for detailed tests. Using the criterion of success in developing to the adult stage, the pattern of tolerance to irradiation in C. illepida was generally eggs < early instars < late instars < pupae. The most tolerant stage potentially occurring in harvested fruits was late (fourth and fifth) instars. Development to adult was reduced slightly in late instars receiving an irradiation dose of 62.5 Gy, whereas development to adult was dramatically reduced in late instars receiving irradiation doses > or = 125 Gy. No C. illepida larvae receiving an irradiation dose > or = 125 Gy emerged as adults and produced viable eggs, indicating sterility can be achieved at doses well below 250 Gy. In large scale tests, when 11,256 late instars were irradiated with a target dose of 250 Gy, 951 pupated (8.4%) and none eclosed as adults. Within the pupal stage, tolerance increased with age; 7- to 8-d-old pupae (the oldest pupae tested) treated with an irradiation dose of 125 Gy produced viable offspring, whereas those treated with a dose of 250 Gy produced no viable offspring. Irradiation of adults with a target dose of 250 Gy before pairing and mating resulted in no viable eggs. Irradiation of actively ovipositing adult females resulted in no subsequent viable eggs. Therefore, the irradiation quarantine treatment of a minimum absorbed dose of 250 Gy approved for Hawaii's fruits will effectively disinfest fruits of any Cryptophlebia in addition to fruit flies.