Sulfuryl fluoride as a quarantine treatment for Anoplophora glabripennis (Coleoptera: Cerambycidae) in regulated wood packing material

The Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), was probably introduced into the United States from China with solid wood packing and dunnage during the 1980s, and it has recently become established in limited infestations near several major cities in the United States. Regulated wood packing material (RWPM) arriving in the United States from China is required to undergo fumigation with methyl bromide (MeBr), to be heat treated, or kiln dried. Sulfuryl fluoride (SF) is a candidate fumigant to replace MeBr under certain conditions. SF fumigations were conducted in 432-liter Lexan chambers held in a 6.1-m (20-foot) refrigerated container for temperature control. Each fumigation consisted of 12 Populus spp. 10- by 10- by 115-cm timbers, of high moisture content, naturally infested with Asian longhorned beetle. During 2001, we fumigated wood for 24 h at a range of doses (20-112 g/m3) and temperatures (4.4, 10.0, 15.6, and 21.1 degrees C) and subjected the data to probit analysis. Confirmatory fumigations were conducted at doses of 120 and 104 g/m3 at temperatures of 10.0 and 15.6 or 21.1 degrees C, respectively, which resulted in complete kill of all larvae. Pupae that became available later in the year as temperatures warmed were fumigated at 15.6 and 21.1 degrees C with 104 g/m3, which resulted in complete pupal mortality. The next year (2002), we conducted 24-h fumigations with doses of 116 g/m3 at 4.4 and 10.0 degrees C with cold-harvested wood infested with cold-acclimated larvae. Cold-acclimated larvae required much higher concentration times time (CxT) product for control at 4.4 and 10.0 degrees C compared with nonacclimated larvae. Sulfuryl fluoride treatments at a dose of 104 g/m3 and temperature of 15.6 degrees C and above and that achieved a CxT product of 1,095 g-h/m3 or above are recommended for RWPM infested with Asian longhorned beetle larvae and pupae.     

Ultrastructural alterations of Erwinia carotovora subsp. atroseptica caused by treatment with aluminum chloride and sodium metabisulfite

Aluminum and bisulfite salts inhibit the growth of several fungi and bacteria, and their application effectively controls potato soft rot caused by Erwinia carotovora. In an effort to understand their inhibitory action, ultrastructural changes in Erwinia carotovora subsp. atroseptica after exposure (0 to 20 min) to different concentrations (0.05, 0.1, and 0.2 M) of these salts were examined by using transmission electron microscopy. Plasma membrane integrity was evaluated by using the SYTOX Green fluorochrome that penetrates only cells with altered membranes. Bacteria exposed to all aluminum chloride concentrations, especially 0.2 M, exhibited loosening of the cell walls, cell wall rupture, cytoplasmic aggregation, and an absence of extracellular vesicles. Sodium metabisulfite caused mainly a retraction of plasma membrane and cellular voids which were more pronounced with increasing concentration. Bacterial mortality was closely associated with SYTOX stain absorption when bacteria were exposed to either a high concentration (0.2 M) of aluminum chloride or prolonged exposure (20 min) to 0.05 M aluminum chloride or to a pH of 2.5. Bacteria exposed to lower concentrations of aluminum chloride (0.05 and 0.1 M) for 10 min or less, or to metabisulfite at all concentrations, did not exhibit significant stain absorption, suggesting that no membrane damage occurred or it was too weak to allow the penetration of the stain into the cell. While mortality caused by aluminum chloride involves membrane damage and subsequent cytoplasmic aggregation, sulfite exerts its effect intracellularly; it is transported across the membrane by free diffusion of molecular SO2 with little damage to the cellular membrane.     

Toxicity, feeding deterrence, and effect of activity of 1,8-cineole from Artemisia annua on progeny production of Tribolium castanaeum (Coleoptera: Tenebrionidae)

1,8-Cineole isolated from Artemisia annua was tested against Tribolium castaneum (Herbst) for contact toxicity, fumigant toxicity, and antifeedant activity. The adults of T. castaneum were more susceptible than larvae to both contact and fumigant toxicity of 1,8-cineole, and LD50 and LC50 values of 108.4 microg/mg body weight of adult insect and 1.52 mg/liter air were found, respectively. Furthermore, 14-d-old T. castaneum larvae were more tolerant than 16- and 18-d-old larvae and adults to the contact toxicity of 1,8-cineole, but the 16- and 18-d-old larvae have similar susceptibility. In contrast, all the larvae (14-18 d old) of T. castaneum were much more tolerant than the adults to the fumigant action, but larvae of different ages had similar susceptibility. The compound 1,8-ciineole applied to filter paper at a concentration of 3.22-16.10 mg/cm2 significantly (P < 0.05) reduced the hatching of T. castaneum eggs and the subsequent survival rate of the larvae. Adult emergence was also reduced by 1,8-cineole. Feeding deterrence of 81.9% was achieved in T. castateum adults by using a concentration of 121.9 mg/g food, whereas larvae showed 68.8% at the same concentration.     

Approved quarantine treatment for Hessian fly (Diptera: Cecidomyiidae) in large-size hay bales and Hessian fly and cereal leaf beetle (Coleoptera: Chrysomelidae) control by bale compression

A quarantine treatment using bale compression (32 kg/cm2 pressure) and phosphine fumigation (61 g/28.3 m3 aluminum phosphide for 7 d at 20 degrees C) was approved to control Hessian fly, Mayetiola destructor (Say), in large-size, polypropylene fabric-wrapped bales exported from the western states to Japan. No Hessian fly puparia (45,366) survived to the adult stage in infested wheat, Triticum aestivum L., seedlings exposed to the treatment in a large-scale commercial test. Daily temperatures (mean +/- SEM) inside and among bales in three test freight containers were 17.8 +/- 0.2 front top, 17.0 +/- 0.2 front bottom, 17.3 +/- 0.2 middle bale, 15.7 +/- 0.3 middle air, 18.5 +/- 0.1 back top, and 18.1 +/- 0.1 degrees C back bottom, allowing the fumigation temperature to be established at > or = 20 degrees C. Mean fumigant concentrations ranged from 208 to 340 ppm during the first 3 d and ranged from 328 to 461 ppm after 7 d of fumigation. Copper plate corrosion values inside the doors, and in the middle of the large-size bales in all locations indicated moderate exposure to hydrogen phosphide (PH3). PH3 residues were below the U.S. Environmental Protection Agency tolerance of 0.1 ppm in animal feeds. The research was approved by Japan and U.S. regulatory agencies, and regulations were implemented on 20 May 2005. Compression in large-size bale compressors resulted in 3-3.6 and 0% survival of Hessian fly puparia and cereal leaf beetle, Oulema melanopus (L.), respectively. Bale compression can be used as a single treatment for cereal leaf beetle and as a component in a systems approach for quarantine control of Hessian fly.     

Ultrastructural Alterations of Erwinia carotovora subsp. atroseptica Caused by Treatment with Aluminum Chloride and Sodium Metabisulfite

Aluminum and bisulfite salts inhibit the growth of several fungi and bacteria, and their application effectively controls potato soft rot caused by Erwinia carotovora. In an effort to understand their inhibitory action, ultrastructural changes in Erwinia carotovora subsp. atroseptica after exposure (0 to 20 min) to different concentrations (0.05, 0.1, and 0.2 M) of these salts were examined by using transmission electron microscopy. Plasma membrane integrity was evaluated by using the SYTOX Green fluorochrome that penetrates only cells with altered membranes. Bacteria exposed to all aluminum chloride concentrations, especially 0.2 M, exhibited loosening of the cell walls, cell wall rupture, cytoplasmic aggregation, and an absence of extracellular vesicles. Sodium metabisulfite caused mainly a retraction of plasma membrane and cellular voids which were more pronounced with increasing concentration. Bacterial mortality was closely associated with SYTOX stain absorption when bacteria were exposed to either a high concentration (0.2 M) of aluminum chloride or prolonged exposure (20 min) to 0.05 M aluminum chloride or to a pH of 2.5. Bacteria exposed to lower concentrations of aluminum chloride (0.05 and 0.1 M) for 10 min or less, or to metabisulfite at all concentrations, did not exhibit significant stain absorption, suggesting that no membrane damage occurred or it was too weak to allow the penetration of the stain into the cell. While mortality caused by aluminum chloride involves membrane damage and subsequent cytoplasmic aggregation, sulfite exerts its effect intracellularly; it is transported across the membrane by free diffusion of molecular SO₂ with little damage to the cellular membrane.     

Bale compression and hydrogen phosphide fumigation to control cereal leaf beetle (Coleoptera: Chrysomelidae) in exported rye straw

Control of larvae and adults of cereal leaf beetle, Oulema melanopus (L.), by bale compression and hydrogen phosphide fumigation was studied in rye straw, Secale cereale L., in Aurora, OR. Natural mortality of larvae after transport was 4.0 +/- 1.0% (mean +/- SEM). Compression (105 kg/cm2) of larvae in standard bales (122 cm long) of rye straw resulted in 100% mortality. Compression of adults in standard bales plus storage of the compressed bales (56 cm long) for 1 d in a freight container resulted in 100% mortality. A KD50 of 102 ppm hydrogen phosphide for 1 h was estimated from the probit regression line developed from dose-response data at 21 degrees C in basic laboratory tests. The LD50s and LD99s were 163 and 6,910 ppm for 2-h exposures and were 18 and 42 ppm for 6-h exposures at 21 degrees C, respectively. A tested close of 400 ppm for 24 h at 21 degrees C resulted in 100% mortality of the adults. Larvae (n = 10,560) and adults (n = 18,602) did not survive exposure to bale compression followed by hydrogen phosphide fumigation (60 g/28.3 m3) for 3 d in rye straw loaded in freight containers in large-scale tests. Copper plate corrosion values indicating the severity of exposure to hydrogen phosphide were 13 and 12, and mean temperatures of five locations in the freight container were 25 and 26 degrees C in large-scale tests with the larvae and adults, respectively. Hydrogen phosphide concentrations were > or = 400 ppm throughout the 3-d fumigation for larvae and during the first day of fumigation for adults. We propose that cereal leaf beetle can be controlled by a single treatment of bale compression followed by a 1-d storage period or by a fumigation in which 400 ppm hydrogen phosphide is maintained for 1 d at 21 degrees C or above. We confirmed that a multiple quarantine treatment of bale compression) followed by a 3-d fumigation will control cereal leafbeetle in exported rye straw.     

Effects of river red gum, Eucalyptus camaldulensis, litter on golden perch, Macquaria ambigua

Aquaria with added river red gum, Eucalyptus camaldulensis , litter became hypoxic, with decreased pH and contained up to 30 mg 1⁻¹ tannin and lignin. Survival of golden perch, Macquaria ambigua , larvae in aquaria treated with a simulated annual litter density of 450 g m⁻² for 72 h was 14·9% for 15-day-old larvae and 0% for 8-day-old larvae. A litter density of 1223 g m⁻² resulted in total mortality for both age groups of larvae. Aeration increased survival of larvae to a minimum of 68·8% in 1223 g m⁻² litter treatments compared to 89·8% in aerated controls and 86·8% in non-aerated controls. A kinetic behavioural assay was used to detect alarm responses in golden perch larvae and juveniles exposed to leachates from river red gum bark, leaves and wood. Eight-day-old larvae exposed to bark and wood leachates (0·001–10 g 1⁻¹) exhibited an initial period of hyperactivity, followed by a concentration-dependent decrease in spontaneous activity. Larvae exposed to leaf leachates displayed only a decrease in spontaneous activity. Four-month-old juveniles exposed to wood leachates were also initially hyperactive, then progressively developed mild hypoactivity at increasing leachate concentrations. Juveniles exposed to wood leachates at 20g 1⁻¹ for 30min suffered 97·5% mortality in 96 h. Wood leachates induced dose-dependent lamellar fusion, epithelial dissociation and necrosis in the gills. The presence of toxic leachates and low oxygen availability in flooded river red gum forests may make these habitats unsuitable as nursery areas for native fish.     

双去甲氧基姜黄素对朱砂叶螨的生物活性及作用方式

在室温26℃±1℃、湿度60%～80%、光照14 h条件下,测定了天然植物活性成分双去甲氧基姜黄素对重要植食性害螨朱砂叶螨各螨态的触杀和熏蒸活性,对幼螨、若螨、成螨的驱避活性,以及对雌成螨的产卵抑制活性.采用玻片浸渍法测得双去甲氧基姜黄素对朱砂叶螨雌成螨48 h的LCso为0.433 mg·mL-1.在0.883 mg·mL-1(LC70)的浓度下,双去甲氧基姜黄素对朱砂叶螨不同螨态触杀活性的大小依次为幼螨＞若螨＞成螨＞卵,其中对幼螨24h和48 h的校正死亡率分别为60.0%和83.3%;对朱砂叶螨各螨态的熏蒸作用不明显,024h和48 h的校正死亡率均小于3%;对幼螨、若螨以及成螨均表现出较强的驱避作用,其中对幼螨的效果最好,不同处理时间的驱避率均在85%以上,其次是若螨,对成螨的驱避性相对较差,72 h的驱避率仅为47.8%.同时对雌成螨有明显的产卵抑制作用,处理后120 h产卵抑制率达到89.3%.表明双去甲氧基姜黄素对朱砂叶螨的主要作用方式为触杀、驱避和产卵抑制作用.     

Comparative evaluation of pyrethroid insecticide formulations against Triatoma infestans (Klug): residual efficacy on four substrates

We investigated the residual efficacy of four insecticide formulations used in Chagas disease vector control campaigns: cyfluthrin 12.5% suspension concentrace (SC), lambda-cyhalothrin 10% wettable powder (WP), deltamethrin 2.5% SC, and 2.5% WP on four types of circular blocks of wood, straw with mud, straw with mud painted with lime, and mud containing 5% of cement. Three concentrations of these insecticides were tested: the LC90 (previously determined on filter paper), the double of the LC90, and the recommended operational dose. For each bioassay test, 15 third-stage nymphs of Triatoma infestans (Klug) (Hemiptera: Reduviidae) were exposed for 120 h to each treatment at 24 h, 30, 60, 90, and 180 days post-spraying. Mortality rates, moulting history and behaviour were recorded at 24, 48, 72, and 120 h of exposure. Mortality rates were highest during the first 30 days post-spraying. Highest mortality rates (above 50%) were observed for deltamethrin 2.5% SC and lambda-cyhalothrin 10% WP on wood blocks up to three months post-spraying. Mud was the substrate on which treatments showed lowest persistence, with the other two substrates showing intermediate residual efficacy of all treatments. During the first 30 days WP formulations were not as effective as SC flowable formulations but, overall in the longer term, WP gave grater mortality rates of T. infestans nymphs exposed at up to six months post-spraying. Porous surfaces, especially mud, showed most variability presumably due to absorption of the insecticide. In contrast the less porous surfaces (i.e. wood and lime-coated mud) kept mortality rates high for longer post-treatment, irrespective of the insecticide concentration used.     

The moisture content of timber in relation to attack by Lyctus powder-post beetles

The relation between moisture content of oak sapwood and rate of development of Lyctus brunneus has been investigated at 23C. Oviposition took place normally in samples of various moisture contents between 5.5 and 23.5%. The eggs hatched in wood of 7.7% moisture content, but not of 61% moisture content, although the embryos developed fully. The rate of gain in weight of young larvae increased to a maximum as the moisture content of the wood rose to about 14.2% and then fell again. The rate of development from egg to adult followed a somewhat similar curve with a maximum at 16% moisture content. At all moisture contents the first male emerged a few days earlier than the first female. The average weight of the male beetles was less and their moisture content greater than those of the corresponding females. The average weight of the beetles showed a tendency to increase at moisture contents of the wood above 97%. The average moisture content of the males was fairly constant up to 156% moisture content of the wood, but that of the females rose slowly as the moisture content of the samples increased. In wood of 15.6% moisture content at 23C. the minimum period of development from egg to adult was 167 days; at 14.2% moisture content and 27.5 C. this was reduced to 142 days. Green timber was shown to be liable to infestation as soon as a superficial skin was formed of a suitable moisture content; the upper limiting moisture content is probably the fibre saturation point. Control of moisture content of timber cannot be achieved sufficiently easily for it to become an important method for the control of Lyctus infestation.     

Control of Codling Moth,Cydia pomonella (Lepidoptera: Tortricidae), with Steinernema carpocapsae: Effects of Supplemental Wetting and Pupation Site on Infection Rate

Infection of cocooned codling moth (cydia pomonella) larvae by the entomopathogenic nematode Steinernema carpocapsae was studied in three field experiments. Factors that varied within or between experiments included method of application, type of substrate containing cocooned larvae, time when nematodes were applied, seasonal effects, and supplemental wetting before or after nematode application. Conventional air-blast sprayer applications of 0.5–5.0 million infective juveniles (IJs)/tree in fall resulted in ca. 30% mortality of larvae in cardboard trap bands, whereas hand-gun application (2 million IJs/tree) produced mortality of ca. 70%. Application in the evening caused higher larval mortality than application in the morning when no supplemental wetting was used after treatments. Morning and evening applications caused equivalent larval mortality when a postwetting treatment was included. In a trial conducted in midsummer, supplemental wetting, either before or after hand-gun application of 1 million IJs/tree, enhanced nematode-produced mortality. Mortality approached 100% if both pre- and postwetting was used. Larvae in exposed cocoons on apple wood were infected at a higher rate (86%) than those on wood in less exposed positions (73%) or in nonperforated cardboard (72%). Mortality rates for larvae in perforated cardboard were intermediate (77%). Application volumes used to deliver nematodes slightly enhanced infection rate of larvae in some substrates but not others. In one trial, parasitism of codling moth by the wasp Mastrus ridibundus (Ichneumonidae) was negatively correlated with nematode infection of codling moth larvae. Dissections showed that ca. 10% of larvae infected by nematodes had been attacked by the wasp.     

Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila

Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg⁹. Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment.     

Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila

Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg⁹. Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment.     

Toxicity of Buprofezin on the Survival of Embryo and Larvae of African Catfish,Clarias gariepinus (Bloch)

Buprofezin is an insect growth regulator and widely used insecticide in Malaysia. The present study evaluated the toxic effects of buprofezin on the embryo and larvae of African catfish (Clarias gariepinus) as a model organism. The embryos and larvae were exposed to 7 different concentrations (0, 0.05, 0.5, 5, 25, 50 and 100 mg/L) of buprofezin. Each concentration was assessed in five replicates. Eggs were artificially fertilized and 200 eggs and larvae were subjected to a static bath treatment for all the concentrations. The mortality of embryos was significantly increased with increasing buprofezin concentrations from 5 to 100 mg/L (p< 0.05). However, the mortality was not significantly different (p<0.05) among the following concentrations: 0 (control), 0.05, 0.5 and 5 mg/L. Data obtained from the buprofezin acute toxicity tests were evaluated using probit analysis. The 24 h LC₅₀ value (with 95% confidence limits) of buprofezin for embryos was estimated to be 6.725 (3.167-15.017) mg/L. The hatching of fish embryos was recorded as 68.8, 68.9, 66.9, 66.4, 26.9, 25.1 and 0.12% in response to 7 different concentrations of buprofezin, respectively. The mortality rate of larvae significantly (p<0.05) increased with increasing buprofezin concentrations exposed to 24-48 h. The 24 and 48 h LC₅₀ values (with 95% confidence limits) of buprofezin for the larvae was estimated to be 5.702 (3.198-8.898) and 4.642 (3.264-6.287) mg/L respectively. There were no significant differences (p>0.05) in the LC₅₀ values obtained at 24 and 48 h exposure times. Malformations were observed when the embryos and larvae exposed to more than 5 mg/L. The results emerged from the study suggest that even the low concentration (5 mg/L) of buprofezin in the aquatic environment may have adverse effect on the early embryonic and larval development of African catfish.     

Toxicity of solvents and surfactants to Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) and Dermacentor nitens (Neumann, 1897) (Acari: Ixodidae) larvae

The objective of this work was to verify the sensitivity of Amblyomma cajennense and Dermacentor nitens larvae to the solvents ethanol, methanol, acetone, xylol and dimethyl sulfoxide (DMSO) and the surfactant Tween 80. The first four solvents were tested at analytical purity while the DMSO and surfactant Tween 80 were tested at a concentration of 1%. The substances tested at analytical purity that caused high mortality were also tested at concentrations of 50%, 25% and 1%. The larval packet test was used, with 10 repetitions for each treatment. A control group was also formed with the same number of repetitions, in which the larvae were only exposed to distilled water. In the first experiment, only xylol was highly toxic at the concentrations tested, causing mortality above 90% for larvae of both species. In the second experiment, xylol at 1% and at 25% showed low toxicity to the A. cajennense and D. nitens larvae, respectively, since the percentage mortality was statistically similar to that of the control group (p>0.05).     

Toxicity of ethanedinitrile to Anoplophora glabripennis (Coleoptera: Cerambycidae) larvae

The mortality of naked Anoplophora glabripennis Motschulsky (Coleoptera: Cerambycidae) larvae exposed to ethanedinitrile (cyanogen; C2N2) varied with temperature, time of exposure, and dose of ethanedinitrile. The concentration x time (Ct) product of ethanedinitrile over a range of temperatures (4.4, 10.1, 15.6, and 20.1 degrees C) decreased with increasing temperature, for both 3- and 6-h exposures. The Ct products varied with time of exposure at different temperatures. The variations in mortality at different temperatures are described with a slope ratio (Y = [slope]6h/ [slope]3h). At different temperatures, the concentration of ethanedinitrile and the duration of exposure play different roles in killing A. glabripennis larvae. These results suggest the control of A. glabripennis larvae within wood is achievable.     

Toxicity of phosphine to Carposina niponensis (Lepidoptera: Carposinadae) at low temperature

Carposina niponensis Matsumura (Lepidoptera: Carposinadae), is widely distributed in pome fruit production areas in China and presents a problem in some export markets because it is considered a quarantine pest by some countries. Methyl bromide is the only fumigant used for fumigation of apples (Malus spp.) for export. However, phosphine is a candidate replacement that can be applied directly at low temperature. Here, laboratory tests showed that tolerance of different stages of C. niponensis to phosphine fumigation at 0 degrees C differed greatly; first-second-instar larvae were the least tolerant stage and the mature fifth instars were the most tolerant stage. In the mature larvae, fumigation tests, with a range of phosphine concentrations from 0.42 to 1.95 mg/liters and exposure periods of 24 h to 14 d at 0 degrees C indicated narcosis when phosphine concentration was > or = 1.67 mg/liter and that a 15.52-8.14-d fumigation period was required to achieve 99% mortality with different phosphine concentrations. The expression of C(0.7)T = k was obtained, which indicated that exposure time was much more important than concentration of phosphine in mortality of mature larvae of C. niponensis. All results suggested that phosphine fumigation at low temperature offers promising control of C. niponensis infestation in pome fruit.     

Effect of d-limonene on three stored-product beetles

d-Limonene was investigated for contact and fumigant toxicity, ovicidal effects, oviposition-deterrent, development inhibition, and feeding-deterrent activities against three stored-product beetles (Coleoptera): lesser grain borer, Rhyzopertha dominica (F.); rice weevil, Sitophilus oryzae (L.); and red flour beetle, Tribolium castaneum (Herbst). Contact and fumigant toxicity decreased as larvae aged. Contact toxicity was similar for adults of the three species tested, but R. dominica was most susceptible to fumigant activity. T. castaneum oviposition decreased as concentration of d-limonene increased and d-limonene reduced oviposition up to 92.3% at the concentration of 2.14 mg/cm2. Hatching of d-limonene-treated eggs of T. castaneum was reduced by 94.5% with no subsequent larval and adult survival at 2.14 mg/cm2 concentration. A flour disc bioassay indicated 87.7 to 96.8% feeding-deterrency by d-limonene toward all three insect species tested at the highest concentration of 60.0 mg/g food. These results suggest that d-limonene can be effectively used to suppress populations of stored-product beetles.     

烟草甲细胞色素P450还原酶基因的分子特征与表达模式分析

烟草甲Lasioderma serricorne是一种重要的仓储害虫,长期化学防治导致烟草甲已对多种传统熏蒸剂产生抗性,但其对新型熏蒸剂甲酸乙酯仍处于敏感水平。因此明确其体内细胞色素P450还原酶（cytochrome P450 reductase, CPR）对甲酸乙酯的代谢解毒功能,对开展该药剂的抗性监测及延缓抗性的发生发展具有重要意义。本研究旨在克隆烟草甲LsCPR基因,分析其分子特征和表达特性,为进一步明确其在烟草甲对甲酸乙酯解毒代谢过程中的作用奠定基础。利用RT-PCR技术克隆烟草甲LsCPR基因的开放阅读框（open reading frame, ORF）;利用生物信息学软件分析LsCPR编码蛋白的结构、特征和系统进化关系。采用实时定量PCR技术检测LsCPR在烟草甲不同发育阶段（低龄幼虫、高龄幼虫、蛹、低龄成虫、高龄成虫）、幼虫不同组织（表皮、肠道、脂肪体和马氏管）以及甲酸乙酯熏蒸胁迫后的表达模式。烟草甲LsCPR基因的ORF为2 046 bp（GenBank登录号：MZ423209）,编码681个氨基酸,具有典型的昆虫CPR基因FMN区域、NADPH区域和FAD等保守结构域。系统发育分析表明,烟草甲LsCPR与鞘翅目昆虫聚为一支,且与赤拟谷盗Tribolium castaneum CPR亲缘关系最近。LsCPR在烟草甲不同发育阶段均有表达,在高龄幼虫期的表达水平较高;在幼虫体内的表达部位主要在肠道,其次为脂肪体和马氏管,而在表皮的表达水平最低。LC10、LC30和LC50 3种浓度的甲酸乙酯处理24 h后,烟草甲LsCPR表达量随着胁迫浓度升高而上调且显著高于对照;甲酸乙酯LC50处理烟草甲不同时间（3、6、12、24和48 h）后,LsCPR基因上调表达,24 h时达到表达高峰。推测LsCPR是参与烟草甲代谢甲酸乙酯的候选基因。     

A sampling plan for population dynamics studies on currant clearwing,Synanthedon tipuliformis (Lepidoptera: Sesiidae)

Abstract

Sampling techniques for eggs, larvae, and pupae of the currant clearwing, Synanthedon tipuliformis (Clerck), attacking blackcurrant were developed, and their precision was evaluated. Reliable population estimates for the egg and early larval stages (standard error < 10%) were obtained by sampling one cane with at least two age groups of wood from the north and south halves of seven bushes in nine blocks. Similar precision was obtained for post-winter larvae from samples comprising three canes from each half of the bushes, but using this sampling scheme a lower precision (S.E.?25–30%) had to be accepted for overwintering mortality factors. The pupal population was estimated from the numbers of live larvae at the end of the post-winter feeding stage, and cross-checked from the number of emergence holes in the 1-year-old wood taken for egg sampling the following year. This procedure gave estimates of similar reliability to those for the egg and larval stages.