Chemical control of the Asian citrus psyllid and of huanglongbing disease in citrus.

By 2014, huanglongbing (HLB), the most destructive disease of citrus, and its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri (Kuwayama), became established in all major citrus-growing regions of the world, including the United States, with the exception of California. At present, application of insecticides is the most widely followed option for reducing ACP populations, while application of antibiotics for suppressing HLB disease/symptoms is being practiced in some citrus-growing regions. Application of insecticides during the dormant winter season, along with cultivation of HLB-free seedlings and early detection and removal of symptomatic and asymptomatic trees, has been very effective in managing ACP. Area-wide management of ACP by application of insecticides at low volume in large areas of citrus cultivation has been shown to be effective in managing HLB and reducing management costs. As insecticide resistance is a major problem in sustainable management of ACP, rotation/alternation of insecticides with different chemistries and modes of action needs to be followed. Besides control of the insect vector, use of antibiotics has temporarily suppressed the symptoms of HLB in diseased trees. Recent efforts to discover and screen existing as well as new compounds for their antibiotic and antimicrobial activities have identified some promising molecules for HLB control. There is an urgent need to find a sustainable solution to the HLB menace through chemical control of ACP populations and within HLB-infected trees through the judicious use of labeled insecticides (existing and novel chemistries) and antibiotics in area-wide management programs with due consideration to the insecticide resistance problem.

Neurotoxicology of bis(n)-tacrines on Blattella germanica and Drosophila melanogaster acetylcholinesterase.

A series of bis(n)-tacrines were used as pharmacological probes of the acetylcholinesterase (AChE) catalytic and peripheral sites of Blattella germanica and Drosophila melanogaster, which express AChE-1 and AChE-2 isoforms, respectively. In general, the potency of bis(n)-tacrines was greater in D. melanogaster AChE (DmAChE) than in B. germanica AChE (BgAChE). The change in potency with tether length was high in DmAChE and low in BgAChE, associated with 90-fold and 5.2-fold maximal potency gain, respectively, compared to the tacrine monomer. The optimal tether length for Blattella was 8 carbons and for Drosophila was 10 carbons. The two species differed by only about twofold in their sensitivity to tacrine monomer, indicating that differential potency occurred among dimeric bis(n)-tacrines due to structural differences in the peripheral site. Multiple sequence alignment and in silico homology modeling suggest that aromatic residues of DmAChE confer higher affinity binding, and the lack of same at the BgAChE peripheral site may account, at least in part, to the greater overall sensitivity of DmAChE to bis(n)-tacrines, as reflected by in vitro assay data. Topical and injection assays in cockroaches found minimal toxicity of bis(n)-tacrines. Electrophysiological studies on D. melanogaster central nervous system showed that dimeric tacrines do not readily cross the blood brain barrier, explaining the observed nonlethality to insects. Although the bis(n)-tacrines were not good insecticide candidates, the information obtained in this study should aid in the design of selective bivalent ligands targeting insect, pests, and disease vectors.

Performance of pre-commercial release formulations of spinosad against five stored-product insect species on four stored commodities.

Two liquid and one dry pre-commercial release spinosad formulations were evaluated at the labeled rate of 1 ppm against five stored-grain insect species on wheat, short-grain rice, long-grain rice, and maize. Except on maize, efficacy of spinosad was compared with a currently registered grain protectant, chlorpyrifos-methyl (3 ppm) plus deltamethrin (0.5 ppm). The 7- and 14-day mortalities of the lesser grain borer, Rhyzopertha dominica, were 99.0-100.0% on spinosad and chlorpyrifos-methyl plus deltamethrin-treated wheat, short-grain rice, and long-grain rice. Adult progeny of R. dominica after 42 days on these commodities decreased by 99.7-100.0% relative to progeny production on untreated wheat. Mortality and reduction in adult progeny of the rice weevil, Sitophilus oryzae, on the three commodities, and that of the maize weevil, Sitophilus zeamais, on maize and the red flour beetle, Tribolium castaneum, on wheat were 100.0% only with chlorpyrifos-methyl plus deltamethrin. The liquid spinosad formulations were most effective against the Indianmeal moth, Plodia interpunctella, on maize and wheat. Except for R. dominica, the effectiveness of spinosad on the other species varied with the formulation, exposure time, and commodity. Chlorpyrifos-methyl plus deltamethrin was effective against insect species on the commodities tested.

Effects of pymetrozine, an antifeedant of Hemiptera, on Asian citrus psyllid, Diaphorina citri, feeding behavior, survival and transmission of Candidatus Liberibacter asiaticus.

BACKGROUND: Vector-borne plant disease management can be enhanced by deployment of antifeedants in addition to the use of broad-spectrum neurotoxic insecticides. The effects of pymetrozine on Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, feeding behaviour, survival and transmission of Candidatus Liberibacter asiaticus (Las), the presumed causal pathogen of huanglongbing, were investigated. RESULTS: Pymetrozine applied at 52 and 104 µg mL(-1) to citrus plants [Swingle citrumelo (X Citroncirus webberi Ingram and Moore)] modified the feeding behavior of ACP and increased the amount of time spent performing non-penetration behaviors while decreasing the time spent performing ingestion behaviors compared with the controls 1 day after treatment. However, the antifeedant effect of pymetrozine subsided 5 days after application. Pymetrozine reduced the survival of both adults and nymphs on treated plants compared with the control. However, it had a greater impact on survival of nymphs than on survival of adults. Pymetrozine applied at 52 and 104 µg mL(-1) on Las-infected 'Valencia' sweet orange plants [Citrus sinensis L. (Osbeck)] reduced acquisition (12 and 21% respectively) and transmission (11 and 18% respectively) of Las by feeding ACP adults compared with the controls; however, these reductions were not statistically significant. CONCLUSIONS: Pymetrozine exhibited moderate antifeedant effects by modifying the feeding behavior of ACP adults with short residual activity. The impact of pymetrozine on survival of nymphs was greater than on adults at the higher concentrations tested. Pymetrozine also reduced the acquisition and transmission of Las by feeding ACP adults up to 21 and 18%, respectively, compared with untreated controls.

Effect of pyriproxyfen, a juvenile hormone mimic, on egg hatch, nymph development, adult emergence and reproduction of the Asian citrus psyllid, Diaphorina citri Kuwayama.

BACKGROUND: The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is a vector of bacteria presumably responsible for huanglongbing (HLB) disease in citrus. In this laboratory study, an investigation was made of the activity of pyriproxyfen, a juvenile hormone mimic, on ACP eggs, nymphs and adults to evaluate its potential as a biorational insecticide for inclusion in an integrated pest management (IPM) program for ACP. RESULTS: Irrespective of egg age, timing or method of treatment, a significantly lower percentage of eggs (5-29%) hatched after exposure to 64 and 128 microg mL(-1) of pyriproxyfen. Only 0-36% of early instars (first, second and third) and 25-74% of late instars (fourth and fifth) survived to adults following exposure to 16, 32 and 64 microg mL(-1) of pyriproxyfen. However, 15-20% of adults that emerged following treatment as late instars exhibited morphological abnormalities. Furthermore, pyriproxyfen adversely affected reproduction (fecundity and fertility) of adults that emerged from treated fifth instars or that were treated topically with 0.04 microg as adults. CONCLUSIONS: Application of pyriproxyfen at 64 microg mL(-1) resulted in greater inhibition of egg hatch and suppression of adult emergence compared with lower rates. Pyriproxyfen also markedly reduced female fecundity and egg viability for adults that were exposed either as fifth instars or as newly emerged adults. The ovicidal, larvicidal and reproductive effects against ACP suggest that pyriproxyfen is suitable for integration into an IPM program for ACP.

Quantifying dispersal of Diaphorina citri (Hemiptera: Psyllidae) by immunomarking and potential impact of unmanaged groves on commercial citrus management.

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important pest of citrus. It is an efficient vector of three bacterial pathogens that are the presumptive causal agents of huanglongbing (HLB) or citrus greening disease. The movement patterns and dispersal capabilities of D. citri require study to better understand the spread of HLB and to improve management strategies for D. citri. A recently developed immunomarking technique that uses crude food proteins (chicken egg albumin, bovine casein, and soy protein) was evaluated for marking and tracking movement of D. citri in Florida citrus groves. In general, both egg and milk protein markers exhibited longer residual activity (35 d) than the soy protein marker (20 d) when applied to citrus leaves with a residual activity order of egg > milk > soy protein. However, residues of all three protein markers decreased with a simulated rain; this was more pronounced for soy protein than for egg and milk proteins. Temperature did not significantly affect acquisition of markers by adult D. citri. Egg, milk, and soy protein markers were detected on >90% of adult D. citri for up to 10, 10, and 5 d, respectively, after field application. Addition of tetrasodium ethylenediamine tetraacetic acid (water softener) and/or Silwet L-77 (wetting agent) to marker solutions did not affect longevity of detection. Each of the protein markers was detected on > or =80% of exposed D. citri for up to 30 d after direct application to adults. A field study was conducted to measure movement of D. citri between replicated pairs of 0.4 ha managed and unmanaged citrus plots separated by 60-100 m. Approximately 70% of captured D. citri were found marked 3 d after application of proteins in the field. Using two marker proteins, it was determined that D. citri moved bi-directionally between managed and unmanaged (abandoned) groves within 3 d with a greater number of D. citri adults moving from unmanaged into managed plots than from managed into unmanaged plots (net movement). These data indicate frequent movement by adult D. citri between groves and suggest that unmanaged groves may act as refuge sites for D citri, leading to reinfestation of nearby managed groves.

Antifeedant and sublethal effects of imidacloprid on Asian citrus psyllid, Diaphorina citri.

BACKGROUND: Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, transmits the causal bacteria of the devastating citrus disease huanglongbing (HLB). Because of the variation in spatial and temporal uptake and systemic distribution of imidacloprid applied to citrus trees and its degradation over time in citrus trees, ACP adults and nymphs are exposed to concentrations that may not cause immediate mortality but rather sublethal effects. The objective of this laboratory study was to determine the effects of sublethal concentrations of imidacloprid on ACP life stages. RESULTS: Feeding by ACP adults and nymphs on plants treated daily with a sublethal concentration (0.1 microg mL(-1)) of imidacloprid significantly decreased adult longevity (8 days), fecundity (33%) and fertility (6%), as well as nymph survival (12%) and developmental rate compared with untreated controls. The magnitude of these negative effects was directly related to exposure duration and concentration. Furthermore, ACP adults that fed on citrus leaves treated systemically with lethal and sublethal concentrations of imidacloprid excreted significantly less honeydew (7-94%) compared with controls in a concentration-dependent manner suggesting antifeedant activity of imidacloprid. CONCLUSIONS: Sublethal concentrations of imidacloprid negatively affect development, reproduction, survival and longevity of ACP, which likely contributes to population reductions over time. Also, reduced feeding by ACP adults on plants treated with sublethal concentrations of imidacloprid may potentially decrease the capacity of ACP to successfully acquire and transmit the HLB causal pathogen.

Influence of posttreatment temperature on the toxicity of insecticides against Diaphorina citri (Hemiptera: Psyllidae).

The psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is one of the most important pests of citrus worldwide because it efficiently vectors three bacteria in the genus Candidatus Liberibacter that cause the devastating citrus greening disease (huanglongbing). Current management practices for this insect pest rely on multiple sprays of foliar insecticides and one or two applications of soil systemic insecticides per season. Effective psyllid and disease management in Florida requires insecticide applications throughout the entire season over wide ranging temperature and environmental conditions. Using a petri dish bioassay technique, the effect of posttreatment temperature (range, 17-37 degrees C) on the toxicity of selected organophosphate (chlorpyrifos and dimethoate), carbamate (carbaryl), avermectin (abamectin), pyrethroid (bifenthrin, zeta-cypermethrin, fenpropathrin, and lambda-cyhalothrin), and neonicotinoid (acetamiprid, imidacloprid, and thiamethoxam) insecticides was evaluated against adult D. citri. The toxicity of both organophosphates showed a positive temperature correlation within the 17-37 degrees C range. Similarly, carbaryl (carbamate) and abamectin (avermectin) exhibited increased toxicity with increasing temperature from 17 to 37 degrees C, with abamectin showing higher overall temperature-dependent toxicity against D. citri adults than carbaryl. With the exception of bifenthrin, which showed a positive temperature-dependent toxicity correlation between 27 and 37 degrees C, all other pyrethroids tested exhibited a negative correlation over the temperature range examined. The toxicity of fenpropathrin and lambda-cyhalothrin dramatically decreased with increasing temperature from 17 to 37 degrees C. The neonicotinoids imidacloprid and thiamethoxam exhibited a mixed response to increasing temperature, whereas acetamiprid showed a positive temperature correlation. However, all three neonicotinoids showed positive temperature-dependent toxicity correlations against D. citri adults over the temperature range tested. These data will enable citrus growers to choose the most effective insecticides for D. citri control from the various classes currently available depending on the prevailing temperature conditions.

Toxicity and disruption of midgut physiology in larvae of the European corn borer, Ostrinia nubilalis, by anion transporter blockers.

In this study, four blockers of anion transporters (ATs) belonging to four different classes of organic acids, including DIDS (4, 4'-diisothiocyanatostilbene-2, 2'- disulfonic acid; a stilbene disulfonic acid), NPPB [(5-nitro-2-(3-phenylpropylamino) benzoic acid; an anthranilic acid)], 9-AC (anthracene-9-carboxylic acid; an aromatic carboxylic acid), and IAA-94 (indanyloxy acetic acid; an indanyloxy alkanoic acid), were tested for their toxicity against the European corn borer (ECB), Ostrinia nubilalis. All the AT blockers inhibited the growth of larvae, increased the developmental time, and decreased survival compared to controls, when second-instar ECB larvae were fed for seven days on treated diet. In general, DIDS and NPPB were the most active compounds, with the rank order of activity being DIDS>NPPB>IAA-94>9-AC. All the AT blockers decreased the midgut alkalinity in fifth-instar larvae when fed for 3 h on treated diet. Effective concentrations required for 50% decrease in midgut alkalinity (EC(50)) ranged between 29.1 and 41.2 ppm and the rank order of activity was NPPB>DIDS>IAA-94>9-AC. Similarly, all the tested AT blockers inhibited (36)Cl(-) uptake from the midgut lumen in fifth-instar larvae when fed for 3 h on treated diet. Concentrations required for 50% inhibition of (36)Cl(-) uptake (IC(50)) ranged between 7.4 and 11.0 ppm and the rank order of activity was DIDS>NPPB>9-AC >IAA-94. Modest to highly strong positive correlations observed among growth, midgut alkalinity, and midgut Cl(-) ion transport in AT blocker-fed larvae suggested that these effects are causally related to each other. Finally, AT blockers have the potential to become good candidates for development of insecticides with a unique mode of action.

Dynamic model for predicting survival of mature larvae of Tribolium confusum during facility heat treatments.

Structural heat treatment, a viable alternative to methyl bromide fumigation, involves raising the ambient temperature of food-processing facilities between 50 and 60 degrees C by using gas, electric, or steam heaters, and holding these elevated temperatures for 24 h or longer to kill stored-product insects. A dynamic model was developed to predict survival of mature larvae, which is the most heat-tolerant stage of the confused flour beetle, Tribolium confusum (Jacquelin du Val), at elevated temperatures between 46 and 60 degrees C. The model is based on two nonlinear relationships: 1) logarithmic survival of T. confusum mature larvae as a function of time, and 2) logarithmic reduction in larval survival as a function of temperature. The dynamic model was validated with nine independent data sets collected during actual facility heat treatments conducted on two separate occasions at the Kansas State University pilot flour and feed mills. The rate of increase of temperature over time varied among the nine locations where mature larvae of T. confusum were exposed, and the approximate heating rates during the entire heat treatment ranged from 1.1 to 13.2 degrees C/h. The absolute deviation in the predicted number of larvae surviving the heat treatment was within 3-7% of the actual observed data. Comparison of the absolute deviation in the time taken for equivalent larval survival showed that the model predictions were within 2-6% of the observed data. The dynamic model can be used to predict survival of mature larvae of T. confusum during heat treatments of food-processing facilities based on time-dependent temperature profiles obtained at any given location.

Morphological characterization of the antennal sensilla of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), with reference to their probable functions.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is one of the most significant economic pests of citrus worldwide. This insect vectors three phloem-restricted bacteria in the genus Candidatus Liberibacter that cause huanglongbing (citrus greening), the most severe disease limiting citrus production worldwide. We examined the external morphology of the antennal sensilla of male and female D. citri using scanning electron microscopy (SEM) and determined the putative functions of the identified sensilla using transmission electron microscopic (TEM) techniques. The filiform antennae of D. citri were of the conventional type comprised of a basal scape, pedicel and a long, thread-like flagellum, which is composed of eight flagellomeres. Eleven morphologically unique sensillar types were found and described on the antennae of male and female D. citri. Of those identified, the two apical setae, multiporous types I and II sensilla trichoidea, and the antennal rhinaria were porous and may be involved in perception of host- and mate-related volatile chemicals. However, the aporous types I, II and III sensilla trichoidea may have mechanosensory functions and the chaetica sensilla, cavity sensilla and unidentified uniporous sensilla may be involved in proprioception, thermo-hygroreception and cold detection, respectively. The shape, external morphology and array of sensilla on the antennae of male and female D. citri were similar. The only major difference observed was in the morphology of the short apical setae, whose tips were recessed inward in females but not so in males. The results are discussed in relation to plausible roles of the identified sensilla in mate and host location by this species.

Nematicidal activity of anion transport blockers against Meloidogyne incognita, Caenorhabditis elegans and Heterorhabditis bacteriophora.

BACKGROUND: Because methyl bromide has been phased out as a soil sterilant, new nematicides are urgently needed. Four different chemical classes of organic acids acting as anion transport (AT) blockers were tested against a free-living nematode, Caenorhabditis elegans Maupas, a plant-parasitic nematode, Meloidogyne incognita (Kofoid and White) Chitwood, and an entomopathogenic nematode, Heterorhabditis bacteriophora Poinar, in toxicity bioassays. The materials tested were DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid), 9-AC (anthracene-9-carboxylic acid), NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] and IAA-94 (indanyloxyacetic acid). RESULTS: All the compounds showed slowly developing nematicidal activity against second-stage juveniles of M. incognita and adults of C. elegans, but not against H. bacteriophora infective-stage juveniles. The LC(50) values of these compounds were < 50 mg L(-1) after 48 and 72 h incubation, while at 168 h incubation the LC(50) values were < 10 mg L(-1) for both sensitive species. Across both species and time, the LC(50) values generally differed no more than twofold among the four compounds tested in this study. In contrast, none of the compounds (200 mg L(-1)) caused more than control mortality to H. bacteriophora, even after 168 h of incubation. CONCLUSION: These compounds are potential leads for commercial nematicides. The insensitivity to H. bacteriophora is consistent with the natural exposure of this nematode to DST (3,5-dihydroxy-4-isopropylstilbene), a stilbene produced by its symbiotic bacterium. Based on the known activity of the compounds used in this study, it is suggested that anion transporters form the probable target sites for DIDS, 9-AC, NPPB and IAA-94 in nematodes.