Nano essential oils against the red palm weevil,Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

The red palm weevil, Rhynchophorus ferrugineus, is the biggest pest of various palm species. The aim of this study was to evaluate the impact of the essential oils of purslane, mustard and castor (bulk and nano) on R. ferrugineus and to evaluate the oviposition deterrent effect to prevent new infestations. Prepared concentrations of essential oils (bulk and nano) were tested for their impact on R. ferrugineus larvae, pupae and adults. The most effective oil was purslane oil (bulk and nano), followed by mustard and the least effective was castor. The percent mortality of larvae was 75.2, 45.3 and 17.9% (bulk phase) and 92.5, 84.4 and 65.5% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The percent mortality of adults was 67.2, 40.0 and 15.6% (bulk phase) and 83.5, 72.9 and 60.0% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The number of eggs laid decreased with increasing concentration of the tested oils. Purslane essential oil showed good impact on R. ferrugineus larvae and adults followed by mustard and castor oils as bulk and as nano.     

上海地区红棕象甲的耐寒性研究

红棕象甲Rhynchophorus ferrugineus (Olivier)是我国危害棕榈科植物的重要入侵害虫, 为探明其越冬抗寒性, 利用过冷却点测定仪测定了红棕象甲上海种群不同虫态的过冷却点, 在低温箱内测定了该虫不同虫态的耐寒性, 然后结合田间越冬模拟试验、气象资料和寄主分布情况, 初步确定了红棕象甲在我国的越冬北界。结果表明： 红棕象甲过冷却点随虫态的发育程度的升高而下降, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫的平均过冷却点分别为-5.92, -6.42, -7.19, -7.43和-11.84℃, 过冷却点由高到低的顺序依次为: 卵>幼虫>成虫。在6, 24, 48和72 h 4个时间处理下, 各虫态在低温与存活率之间呈显著或极显著的logistic回归, 半致死温度（Ltemp₅₀）均随处理时间的延长而上升, 不同虫态在处理72 h 后, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫之间的Ltemp₅₀分别为1.61, -1.67, -2.39, -2.40和-0.40℃, 各虫态耐寒性由弱到强的顺序依次为: 卵<成虫<幼虫。红棕象甲不同发育阶段的过冷却点与其耐寒性并不完全相关, 幼虫和成虫均可能是该虫的越冬虫态。连续两年的田间模拟越冬试验表明, 在上海地区, 红棕象甲的幼虫和成虫的越冬存活率均在60％以上, 说明红棕象甲在上海地区是可以越冬的。根据这些结果, 结合寄主分布情况, 初步将红棕象甲在我国的越冬北界定于北纬35°附近, 即1月份0℃等温线左右。     

Identification and enzymatic characterisation of digestive glucosidases from gut of red palm weevil,Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae)

Red palm weevil, Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae) is a serious pest of date worldwide. Thus, damage to palms (almost exclusively to Phoenix canariensis Hort) has been recorded in various places. Thus, the aim of the current study was to investigate two major digestive enzymes of this insect, α-D-glucosidase and β-D-glucosidase. The results showed that α-D-glucosidase and β-D-glucosidase are present in the insect gut mainly in the midgut and hindgut but trace amounts of the both enzymes were found in the foregut. Optimum temperature for α- and β-glucosidases was found to be 50 and 40?°C, respectively, and pH values were 4.0. The activity of glucosidases were increased by NaCl and KCl and inhibited by some compounds such as MgCl₂ and CaCl₂. Kinetic parameters showed that K _m of α and β-D-glucosidases was 3.15 and 4.11?mM, respectively. Therefore, it is concluded that in this insect species, both α-glucosidase and β-glucosidase are active but with different amounts. Understanding of the digestive physiology and glucosidase activity of red palm weevil is important when new management strategies based on interfering in the gut physiology of the insects are devised.     

A large‐scale gene discovery for the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae)

The red palm weevil (RPW; Rhynchophorus ferrugineus) is a devastating pest of palms, prevalent in the Middle East as well as many other regions of the world. Here, we report a large‐scale de novo complementary DNA (cDNA) sequencing effort that acquired ～5 million reads and assembled them into 26 765 contigs from 12 libraries made from samples of different RPW developmental stages based on the Roche/454 GS FLX platform. We annotated these contigs based on the publically available known insect genes and the Tribolium castaneum genome assembly. We find that over 80% of coding sequences (CDS) from the RPW contigs have high‐identity homologs to known proteins with complete CDS. Gene expression analysis shows that the pupa and larval stages have the highest and lowest expression levels, respectively. In addition, we also identified more than 60 000 single nucleotide polymorphisms and 1 200 simple sequence repeat markers. This study provides the first large‐scale cDNA dataset for RPW, a much‐needed resource for future molecular studies.     

Biochemical and ultrastructural changes in the ovaries of red palm weevil,Rhynchophorus ferrugineus (Coleoptera: Curculionidae) following acute imidacloprid poisoning

This study aims to inspect the effects of Imidacloprid (IMI) acute toxicity in the ovaries of the red palm weevil (Rhynchophorus ferrugineus). The LC₅₀ of IMI was determined to be 15.7 ppm. We assessed the biochemical changes resulting from treatment with four IMI concentrations (10, 15, 20 and 30 ppm). IMI reduced acetylcholine esterase activity in a dose-dependent manner. Glutathione (GSH) content decreased with increasing concentration of IMI; however, GSH was not affected and accompanied by an increase in catalase activity with the highest concentration of IMI. Lipid peroxidation increased steadily as IMI concentration increased. Based on the ultrastructural analyses of the follicle cells and the ooplasm of developing oocytes, acute IMI toxicity produced damaged yolk bodies, pale nuclei, and shrinking microvilli. Fractionation of rough endoplasmic reticulum, deformed mitochondria, and autophagic bodies were also observed. In conclusion, these ovarian pathobiochemical changes could be used as biomarkers of acute IMI exposure.     

Integration of entomopathogenic fungi and eco-friendly insecticides for management of red palm weevil,Rhynchophorus ferrugineus (Olivier)

Red palm weevil (Rhynchophorus ferrugineus) is a voracious pest of date palm worldwide. Pakistan ranks sixth in date palm production globally. Losses to date palm plantations in Pakistan sometimes surpass 10%-20%. Most of the traditional management strategies used by farmers have been found insignificant to combat this voracious pest. The entomopathogenic fungi, Beauveria bassiana [QA-3(L) and QA-3(H)] and insecticides, Nitenpyram (Active 10% SL) [NIT (L) and NIT (H)] were applied to larval (2nd, 4th, and 6th), pupal and adult stages of R. ferrugienus. Integration or alone application of fungi with insecticides at different concentration under laboratory conditions. Combined application was depicted additive and synergistic interactions. Contrarily, highest cumulative mortality (100%) was recorded in 2nd instar larvae as compared to later instar larvae at combined application. The maximum pupal and adult mortality remained 89% and 66% respectively after treatment with [QA-3 (H) + NIT (L)]. The combination of B. bassiana at higher concentration whereas Nitenpyram at lower dose was found more lethal to larvae, pupae and adults of R. ferrugineus. This signifies the need of combining B. bassiana and bio-rational insecticides that can reduce the cost of management with least harm to environment and natural enemies.     

Virulence of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against red palm weevil,Rhynchophorus ferrugineus (Olivier)

Nineteen different isolates of the entomopathogenic fungi Beauveria bassiana sensu lato (s.l.) and Metarhizium anisopliae s.l. (Ascomycota: Hypocreales), recovered from different soil samples (field crops, fruit orchards, vegetable fields and forests) and insect cadavers were tested against Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) at two different spore concentrations (1 × 10⁷ and 1 × 10⁸ conidia mL^?1). Three isolates of B. bassiana and two of M. anisopliae gave >88 % larval and >75 % adult mortality of R. ferrugineus on their highest dose rate respectively. More sporulating cadavers (mycosis) resulted from a high dose rate compared to low dose on both life stages of R. ferrugineus. The current study confirmed the lethal action of B. bassiana and M. anisopliae isolates with mortality levels usually directly proportional to the conidial concentration. This study further confirmed that the isolates recovered from R. ferrugineus dead cadavers gave more mortality compared to the other sources. In the virulence assay two isolates of B. bassiana caused the highest percentage of both larval and adult mortality at all exposure intervals which suggest that they may be the most promising for use in sustainable management programs aimed at microbial control in date palm orchards.     

The culturable bacterial community of frass produced by larvae of Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) in the Canary island date palm

Aims: Larvae of the red palm weevil (RPW) Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) feed inside palm stem tissues, making galleries and producing a wet fermenting frass. We characterized the culturable bacteria associated with frass produced by tunnelling larvae inside the Canary island date palms and investigated the role of frass and gut bacteria in plant polymers breakdown. Methods and Results: A culture‐dependent method was used to isolate bacteria from frass and noninfested palm tissues. Bacterial isolates were grouped into operational taxonomic units based on polymorphisms in the ITS‐PCR profiles, and representative isolates were identified by partial sequencing of the 16S rRNA gene. Frass bacteria were dominated by 2,3‐butanediol fermenter Enterobacteriaceae. None of the bacterial isolates was able to degrade cellulose; however, cellulolytic and hemicellulolytic bacteria were isolated from the larval gut enrichment cultures. Conclusions: Frass bacteria are specifically associated with the RPW larvae and might play beneficial roles for RPW, other than nutritional, that deserve further investigations. Breakdown of plant polymers probably occurs inside the larvae digestive system. Significance and Impact of the Study: Frass and gut micro‐organisms of R. ferrugineus should be included in studies of the interactions between RPW, its plant hosts, and its enemies.     

Immune function differences between two color morphs of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae) at different life stages

Several studies demonstrated that in insects cuticle melanism is interrelated with pathogen resistance, as melanin‐based coloration and innate immunity possess similar physiological pathways. For some insects, higher pathogen resistance was observed in darker individuals than in individuals with lighter cuticular coloration. Here, we investigated the difference in immune response between two color morphs (black and red) and between the life stages (pupa and adult) of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae). Here in this study, cuticle thickness, microbial test (antimicrobial activity, phenoloxidase activity, and hemocyte density), and immune‐related gene expression were evaluated at different stages of RPW. Study results revealed that cuticle thickness of black phenotype was thicker than red phenotype at old‐pupa stage, while no significant difference found at adult stage. These results may relate to the development processes of epidermis in different stages of RPW. The results of antimicrobial activity, phenoloxidase (PO) activity, and hemocyte density analyses showed that adults with a red phenotype had stronger pathogen resistance than those with a black phenotype. In addition to antimicrobial activity and PO activity, we tested relative gene expression in the fat body of old pupae. The results of hemolymph antimicrobial analysis showed that old pupae with a red phenotype were significantly different from those with a black phenotype at 12 hr after Staphylococcus aureus injection, suggesting that red phenotype pupae were more sensitive to S. aureus. Examination of gene expression in the fat body also revealed that the red phenotype had a higher immune response than the black phenotype. Our results were inconsistent with the previous conclusion that dark insects had increased immune function, suggesting that the relationship between cuticle pigmentation and immune function in insects was not a direct link. Additional possible factors that are associated with the immune response, such as life‐history, developmental, physiological factors also need to be considered.     

室内饲养红棕象甲的行为观察

报道红棕象甲Rhynchophorus ferrugineus Fab.成虫交配、产卵行为,描述在交配和产卵过程中发生的多种行为。红棕象甲整日均可见其交配行为,但在暗光周期开始0.5h左右明显有一个交配高峰期。其交配持续时间、交配几率不受虫体大小的影响。在无外界因子干扰的条件下,交配持续时间均可达1min。红棕象甲卵为单产,一处1粒,主要是通过口喙蛀孔辅助完成;观察红棕象甲幼虫、蛹的行为,例如取食、争斗、蜕皮、做茧等行为。且随着低龄幼虫向高龄幼虫的过渡,幼虫的取食和钻蛀行为从棕榈植物中心向外围扩散。通过摇动或震动茧后红棕象甲在其中表现的行为,并结合茧的颜色和质地可准确判断茧中存在的3种虫态。     

Lower temperature thresholds for oviposition and egg hatching of the red palm weevil, Rhynchophorus ferrugineus (Coleoptera: Curculionidae), in a Mediterranean climate

The red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), is an economically important pest of palms worldwide. To better understand the phenology of R. ferrugineus populations in the Mediterranean basin, the objective of the present work has been to estimate the effect of temperatures within the range 10-25°C on the reproductive parameters of R. ferrugineus and to determine its lower temperature thresholds for oviposition and egg hatching. Our results confirm that oviposition in R. ferrugineus is strongly affected by temperature. Lower thresholds for oviposition and egg hatching (15.45° and 13.95°C, respectively) are below the mean monthly temperatures registered in winter in most of the northern shore of the Mediterranean basin. Under these circumstances, new palm infestations would be difficult during most of the winter. These results should be taken into account when planning some palm management practices, such as pruning or pesticide treatments, in areas under a Mediterranean climate.     

一种红棕象甲人工饲料及其效果

【目的】为评价一种红棕象甲Rhynchophorus ferrugineus Oliver人工饲料的饲养效果。【方法】以自然寄主大王棕和甘蔗饲养红棕象甲作对照,连续3代饲养红棕象甲,对红棕象甲各发育阶段的生长和发育指标进行调查。【结果】用人工饲料饲养的红棕象甲幼虫历期和蛹历期(40.20 d和33.99 d)显著低于对照组(60.87 d和36.12 d;109.00 d和42.70 d),用人工饲料饲养的红棕象甲幼虫存活率(63.87%)显著高于对照组(37.99%和48.60%)。人工饲料饲养的红棕象甲,在单雌产卵量和卵孵化率方面与对照组没有显著差异,在蛹重和成虫体重方面显著小于对照组,蛹重比对照组减少了25.55%和63.87%,成虫体重比对照组减少了24.18%和35.16%。【结论】该人工饲料可以很好的满足红棕象甲生殖发育的营养需求,且缩短了世代发育历期,适于红棕象甲种群规模化、长期饲养。     

Pseudomonas aeruginosa,a facultative pathogen of red palm weevil,Rhynchophorus ferrugineus

Pseudomonas aeruginosa was identified as a facultative pathogen of red palm weevil. Intra-haemocoelic injection of the pathogen within larvae and pre-pupae was more effective at killing the insects [with a median lethal dose (LD₅₀) of 9×10² to 2×10³ bacteria/insect] than inoculation by force feeding (LD₅₀ of 10⁵ to 4×10⁵ bacteria/insect) or by wading the insects in a suspension of the pathogen (LD₅₀ of 10⁵ to 2×10⁵ bacteria/insect). Injection of 3×10³ bacteria/insect killed 69% of larvae; small larvae were more susceptible (LD₅₀ of 9×10⁵ bacteria/larva) than either larger larvae (LD₅₀ of 10³ bacteria/larva) or pre-pupa. The median time to death of the small larvae following injection of P. aeruginosa was about 6 days but that following force feeding or wading was about 8 days. A secondary invader, Serratia marcescens, had no effect on the pathogenicity of P. aeruginosa but hastened death of larvae by about 3 days.A. Banerjee and T.K. Dangar were with the Central Plantation Crops Research Institute, Regional Station, Kayangulam 690 533, Kerala, India. They are now with the Central Rice Research Institute. Cuttack 753 006, Orissa, IndiaCPCRI research paper no. 870.     

Entomopathogenic fungi-mediated biological control of the red palm weevil Rhynchophorus ferrugineus

The red palm weevil (RPW), Rhynchophorus ferrugineus, is an important pest of palms, and difficult to control by conventional methods. Therefore, microbial control is an alternative strategy for controlling RPW. Herein, a total of 15 entomopathogenic fungi (EPFs) were subjected to primary pathogenicity screening against last stage of RPW larvae. The preliminary data showed that four Beauveria bassiana isolates (JEF-484, 158, 462 and 507) and one Isaria fumosorosea isolate (JEF-014) resulted in 100 % mortality within 5–10 days post inoculation (d.p.i.), respectively. According to the time required for RPW mortality, JEF-484, 158, 462 and 014 were further subjected to bioassays using 10⁷ conidia/ml suspensions by spraying method. Based on the results, JEF-484 showed the highest mortality and shortest LT₅₀ on the last stage of RPW larvae, followed by JEF-158. The two isolates also showed good conidial production and high thermal stability compared to the other isolates. Therefore, JEF-484 and JEF-158 were selected for bioassays against RPW egg and the last larval stage with different concentrations of 10⁵, 10⁶ and 10⁷ conidia/ml conidial suspensions by spraying method. For the bioassay at the egg stage, JEF-158 showed a significantly higher ovicidal effect than JEF-484. In the larval bioassay, both EPF isolates showed a dosage-dependent effect on the RPW larvae. JEF-484 caused higher mortality in RPW larvae than JEF-158. In summary, the combination of the 2 promising EPF isolates might provide an opportunity for the practical microbial control of RPW at different life stages in palm tree fields.     

Functional compounds from the integument of adult red palm weevil Rhynchophorus ferrugineus

This study deals with the analysis of the integument of the red palm weevil Rhynchophorus ferrugineus of both sexes using the GC–MS technique. The results of the study revealed many promising compounds. These include aspidofractinine-3-methanol (kopsinyl alcohol) which was found in the acetone extract of the sternum of females, and 3-buten-2-ol (32-B) which was found in the extracts of sternum and tergum of males. This compound the aggregation pheromone was secreted by males. Additionally, compounds with methoxy groups were found. These may be responsible for insects’ resistance. This study, through separation and identification of these compounds, aims to open a new possibility for their future medical and therapeutic usage.     

Hemocytes of Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) and their response to Saccharomyces cerevisiae and Bacillus thuringiensis

Originally from tropical Asia, the Red Palm Weevil (RPW Rhynchophorus ferrugineus (Olivier)) is the most dangerous and deadly pest of many palm trees, and there have been reports of its recent detection in France, Greece and Italy. At present, emphasis is on the development of integrated pest management based on biological control rather than on chemical insecticides, however the success of both systems is often insufficient. In this regard, RPW appears to be one pest that is very difficult to control. Thus investigations into the natural defences of this curculionid are advisable. RPW hemocytes, the main immunocompetent cells in the insect, are described for the first time. We identified five hemocyte cell types from the hemolymph of R. ferrugineus: plasmatocytes (∼50%), granulocytes (∼35%), prohemocytes (∼8%), oenocytes (∼4%) and spherulocytes (∼3%). SEM observations were also carried out. Some aspects of RPW interaction with non-self organisms, such as Saccharomyces cerevisiae and the entomopathogen bacterium, Bacillus thuringiensis (Bt), are discussed. Plasmatocytes and granulocytes were involved in nodules and capsule formation as well as in the phagocytosis of yeast. The hemocyte response of RPW larvae to sub-lethal doses of commercial products containing Bt was examined. In vivo assays were carried out and Bt in vegetative form was found in the hemolymph. After a diet containing Bt, the number of total hemocytes, mainly plasmatocytes, in the RPW larva hemolymph declined sharply (∼12%) and then remained at a low level, while the number of other circulating cells was almost unchanged.     

警惕外来危险害虫红棕象甲的扩散

红棕象甲Rhynchophorus ferrugineus(Olivier)是棕榈科植物外来危险害虫,该虫原产印度,20世纪80年代扩散到中东,目前在我国主要分布在海南、广西、广东、台湾、云南、西藏(墨脱)等地区,存在进一步扩张的趋势。文章对该虫的地理起源及分布、生物学特性、寄主及防治等方面作了描述。     

Structure and function of the antennal sensilla of the palm weevil Rhynchophorus palmarum (Coleoptera, Curculionidae)

The distribution, fine structure and function of the sensilla present on the antennal club of Rhynchophorus palmarum were studied. No sex dimorphism was observed. Scanning and transmission electron microscopy showed five types of hair-like structures, four of which were evenly distributed on the antennal club. Two types of hair (IV and V) showed wall pores, a characteristic of olfactory sensilla. The antenna numbers 11,190 +/- 3040 type IV and 7360 +/- 1500 type V hairs. Using single sensillum recording, we identified 17 types of olfactory receptor neurons (ORNs) on the basis of their responses to pheromone and host plant odors, triggering synergic behavioral responses. We characterized highly specific and sensitive ORNs tuned to the aggregation pheromone (18% ORNs; 0.01-1 ng response threshold) and to host plant odors such as ethyl acetate, ethanol, acetoin and guaiacol (10% ORNs; 1-10 ng response threshold). Eleven percent of the ORNs were more generalist, responding to several odors with low sensitivity. Nine percent of the ORNs showed a complex pattern of responses, being co-activated by the pheromone and plant odors. This suggests an interaction at the sensory neuron level between pheromone and plant odors, triggering synergic behavioral responses.