Volatile compounds from Liposcelis bostrychophila (Psocoptera: Liposcelididae) and their environment and their effects on settling behaviour

Headspace from above either Liposcelis bostrychophila Badonnel reared on powdered food; insects without food or from powdered food only were collected on five occasions over a period of four weeks. Volatiles were analysed by thermal desorption gas chromatography-mass spectrometry (TD-GCMS) to look for ecologically significant volatile compounds. Hexanoic acid (HA) was the major compound in the insect only and diet only volatiles (insect 25.6 ± 1.6%, diet 32.9 ± 1.13%). Isobutyric acid was the main compound detected in the volatiles from both insects and diet (35.3 ± 6%). Butyric acid (BA) and acetoin (HB) were also present at >2% of the total peak areas. HB was not detected in the insect only volatiles. Bioassays showed that hexanoic acid was the most repellent chemical. Mixtures of compounds representing the proportions of HA, BA, IBA, and HB in the volatiles did not have any prolonged effect on settling or selection by booklice in a two choice bioassay. L. bostrychophila can avoid individual volatile chemicals but the effects can be masked when they are incorporated into mixtures. As some volatile compounds can be detected by booklice they may govern dispersal, but further work is required to clarify their significance.     

储藏物害虫嗜卷书虱对DDVP熏蒸的行为反应与致死剂量

1 引言 嗜卷书虱(Liposcelis bostrychophila)属啮虫目虱啮科。其体形微小,食性复杂,栖息场地多在粮食仓库及食品加工厂、图书馆、标本馆等地。大量发生时可造成严重损失,传播微生物,并以其尸体、排泄物等污染储藏的粮食和其它储藏物品。王进军等调查发现,在我国“双低”和“三低”贮粮的粮仓中,书虱已成为优势种群,对贮粮安全构成了威胁。使用病原真菌和细菌来防治嗜卷书虱,很难达到理想效果,即使采用菊酯类杀虫剂也难以奏效,贮粮熏蒸中普遍使用的溴甲烷(CH_3Br)和磷化氢(PH_3)对嗜卷书虱的防治效果也不很理想。利用昆虫生长调节剂防治嗜卷书虱的研究已有一些报道,但在具体应用上还存在着一些问题。气调措施可应用于控制书虱种群的增长,但书虱的抗气性发展很快,     

Molecular characterization of two nicotinic acetylcholine receptor subunits from Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

Two nicotinic acetylcholine receptor (nAChR) subunit genes, Lbα1 and Lbα8, were isolated and characterized from psocid, Liposcelis bostrychophila Badonnel, using the rapid amplification of cDNA ends (RACE) technique. They are the first two nAChR family members isolated from the insect order of Psocoptera. The full‐length cDNAs of Lbα1 (GenBank accession number: EU871527) and Lbα8 (EU871526) consist of 2,025 and 1,763 nucleotides, respectively, and an open reading frame of 1,644 and 1,608 bp encoding 547 and 535 amino acid proteins, respectively. Both genes have typical features of nAChR family members, though they share only 56% identity in amino acid sequence. The dendrogram generated by the MEGA 3.1 program shows that the protein deduced by Lbα1 had the closest phylogenetic relationship to Agamα1 from Anopheles gambiae and Amelα1 from Apis mellifera, and Lbα8 shares the highest identity with Agamα8 from An. gambiae and Amelα8 from A. mellifera. Quantitative real‐time PCR analysis showed that Lbα1 was expressed 2.03–6.54‐fold higher than Lbα8 at the different developmental stages of L. bostrychophila. The highest expression levels of Lbα1 and Lbα8 were both detected at adult stage and the lowest were at the third and fourth nymphal stages, respectively. There was a stable and relatively low expression level for Lbα1, whereas there was a descending expression pattern for Lbα8 in the 1st through the 4th nymphal stadia. © 2009 Wiley Periodicals Inc.     

Tracing the geographic origin of the cosmopolitan parthenogenetic insect pest Liposcelis bostrychophila (Psocoptera: Liposcelididae)

The randomly amplified polymorphic DNA technique was used to trace the geographic origin of Liposcelis bostrychophila Badonnel populations in Australia from unknown geographic sources internationally. Haplotype (or clonal) diversity was high, with 474 unique haplotypes found from 616 individuals genotyped. Gene diversity estimates (0.10-0.28) and percent polymorphic loci (38.1-88.1%) were moderate to high for most populations. This resulted in genetic distance estimates that ranged from 0.04 to 0.26 and were significantly different for most pairwise population combinations. G ST values for all populations were also moderate (0.04-0.54) and again were significantly different for most pairwise population comparisons. Analysis of molecular variance revealed that the majority of variation was apportioned among individuals within populations regardless of the level at which they were grouped. Gene flow (Nm) was mostly low for all pairwise populations comparisons with an average Nm=1.8. A non-significant negative correlation between genetic distance and geographic distance was found for worldwide populations. In contrast, within Australian populations a significant positive correlation between genetic distance and geographic distance was detected. Genetic relationships explored using unweighted pair group method analysis and non-metric multidimensional scaling indicated a mixed pattern of genetic similarities among all populations. Multiple introductions, from a wide range of international source populations, have obscured the ability to accurately determine the geographic origin of L. bostrychophila in Australia.     

Effects of removal of Cardinium infection on fitness of the stored-product pest Liposcelis bostrychophila (Psocoptera: Liposcelididae)

Endosymbiotic bacteria that potentially influence reproduction and other fitness-related traits of their hosts are widespread in arthropods, and their appeal to researchers' interest is still increasing. In this study, the effects of removal of Cardinium infection on development, survival, and reproduction of Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae) were investigated in the laboratory. The Cardinium-free strain was obtained by the removal of Cardinium infection by using 1% rifampicin treatment on the Cardinium-infected strain (control) for 4 wk, and no Cardinium gene product was detected in this strain throughout the experiment. The results showed that the removal of Cardinium infection had negative effects on fitness of L. bostrychophila. Compared with the control strain, the Cardinium-free strain (both in first [F1] and second [F2] generation) had a similar developmental time, reduced survivorship of immature stages, as well as reduced fecundity, which resulted in much smaller r(m) values. Using r(m) values, the fitness for Cardinium-free F1 and F2 relative to the control was calculated as 0.81 and 0.74, respectively. We concluded that the use of antibiotics combined with heat treatment might be a good control measure for L. bostrychophila.     

Molecular Characterization and Alternative Splicing of a Sodium Channel and DSC1 Ortholog Genes in Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

Alternative splicing greatly contributes to the structural and functional diversity of voltage-gated sodium channels (VGSCs) by generating various isoforms with unique functional and pharmacological properties. Here, we identified a new optional exon 23 located in the linker between domains II and III, and four mutually exclusive exons (exons 27A, 27B, 27C, and 27D) in domains IIIS3 and IIIS4 of the sodium channel of Liposcelis bostrychophila (termed as LbVGSC). This suggested that more alternative splicing phenomena remained to be discovered in VGSCs. Inclusion of exon 27C might lead to generation of non-functional isoforms. Meanwhile, identification of three alternative exons (exons 11, 13A, and 13B), which were located in the linker between domains II and III, indicated that abundant splicing events occurred in the DSC1 ortholog channel of L. bostrychophila (termed as LbSC1). Exons 13A and 13B were generated by intron retention, and the presence of exon 13B relied on the inclusion of exon 13A. Exon 13B was specifically expressed in the embryonic stage and contained an in-frame stop codon, inclusion of which led to generation of truncated proteins with only the first two domains. Additionally, several co-occurring RNA editing events were identified in LbSC1. Furthermore, remarkable similarity between the structure and expression patterns of LbVGSC and LbSC1 were discovered, and a closer evolutionary relationship between VGSCs and DSC1 orthologs was verified. Taken together, the data provided abundant molecular information on VGSC and DSC1 orthologs in L. bostrychophila, a representative Psocoptera storage pest, and insights into the alternative splicing of these two channels.     

Accumulation and utilization of triacylglycerol and polysaccharides in Liposcelis bostrychophila (Psocoptera, Liposcelididae) selected for resistance to carbon dioxide

Abstract: Two populations of the psocid, Liposcelis bostrychophila Badonnel, were exposed to two CO₂-enriched atmospheres (35% CO₂ + 21% O₂, and 55% CO₂ + 21% O₂, balance N₂) for 30 generations. Controls were reared in normal atmospheres. The reserves of triacylglycerol and polysaccharides were evaluated in adults of the two experimental and the control populations in generations F₁₅ and F₃₀. The utilization rate of triacylglycerol and polysaccharides in the CO₂-enriched atmospheres were also determined in generation F₃₀. The results indicated that the reserves of triacylglycerol and polysaccharides increased significantly during selection for CO₂ resistance; the higher the resistance level, the greater the reserves. Exposure of these populations to controlled atmosphere was associated with a steady utilization of the reserves. By contrast, the unselected population responded to controlled atmospheres by accelerated utilization of triacylglycerol and polysaccharides. Comparison of the utilization rates during CO₂ exposure showed that triacylglycerol is the main energy source, and polysaccharides contribute to metabolic energy supply only to a small extent.     

Toxicological and biochemical characterizations of GSTs in Liposcelis bostrychophila Badonnel (Psocop., Liposcelididae)

Abstract: The toxicological and biochemical characteristics of glutathione S‐transferases (GSTs) in the resistant and susceptible strains of Liposcelis bostrychophila were investigated. The two resistant strains were the dichlorvos‐resistant strain (DDVP‐R) and PH₃‐resistant strain (PH₃‐R), and the resistance factors were 22.36 and 4.51, respectively. Compared with their susceptible counterparts, the activities per insect and specific activities of GSTs in DDVP‐R and PH₃‐R were significantly higher. The apparent Michaelis–Menten constant values (K_m) for 1‐chloro‐2,4‐dinitrobenzene (CDNB) were obviously lower in DDVP‐R and PH₃‐R (i.e. lower K_m values, 1.5625 mm for DDVP‐R and 0.6230 mm for PH₃‐R) when compared with their susceptible counterpart (K_m = 3.5520), indicating a higher affinity to the substrate CDNB in resistant strains. In contrast, the catalytic activity of GSTs towards CDNB in the susceptible strain was significantly higher than those in resistant strains. It was noticeable that when reduced glutathione (GSH) was used as substrate, GSTs from resistant strains both indicated a significantly declined affinity. For the catalytic activity of GSTs towards GSH, only the V_max value in DDVP‐R increased significantly compared with that from the susceptible strain, suggesting an overexpression of GST in this resistant strain. The inhibition kinetics of insecticides to GSTs in vitro revealed that dichlorvos and paraoxon possessed excellent inhibition effects on GSTs. The susceptible strain showed higher sensitivity (I₅₀ = 0.9004 mm ) to dichlorvos than DDVP‐R and PH₃‐R (higher I₅₀s, 8.0955 mm for DDVP‐R and 9.3346 mm for PH₃‐R). As for paraoxon, there was a similar situation. The resistant strains both suggested a higher I₅₀ (1.8735 mm for DDVP‐R, and 0.4291 mm for PH₃‐R) compared with the susceptible strain (0.2943 mm ). These suggested that an elevated detoxification ability of GSTs developed in the resistant strains.     

三色书虱体内共生微生物Wolbachia的wsp基因的分子检测

采用常规PCR和巢式PCR方法对三色书虱Liposcelis tricolor体内的共生微生物Wolbachia的wsp基因进行分子检测;通过Wolbachia的通用引物以及A、B亚群引物分别比较了常规PCR和巢式PCR对wsp基因扩增的灵敏性。从三色书虱体内扩增出了610bp的Wolbachia的wsp基因片段,500bp的WolbachiaA亚群的wsp基因片段和450bp的Wolbachia B亚群的wsp基因片段。扩增结果说明三色书虱被A和B两个亚群的Wolbachia混合感染;巢式PCR比常规PCR更为灵敏。     

杀虫剂胁迫下嗜卷书虱和嗜虫书虱能源物质的代谢比较

采用比色法测定嗜卷书虱（Liposcelis bostrychophila）和嗜虫书虱（L．entomophila）体内甘油三酯、多糖和可溶性蛋白的贮存量。结果表明,嗜虫书虱体内3种物质的贮存量均显著高于嗜卷书虱。在毒死蜱和丁硫克百威的胁迫下,嗜虫书虱体内3种物质的变化幅度均大于嗜卷书虱,其中甘油三酯的含量先增高后降低,整体呈现下口抛物线趋势;多糖和可溶性蛋白的消耗率增加。这些说明在毒死蜱和丁硫克百威的作用下,书虱更多地启动了多糖和可溶性蛋白的代谢,降低了对甘油三酯的利用,以应对杀虫剂的胁迫。     

Inhibition kinetics on carboxylesterase and acetylcholinesterase of Liposcelis bostrychophila and Liposcelis entomophila (Psocop., Liposcelididae) of two insecticides

Abstract: The inhibition kinetics on carboxylesterase (CarE) and acetylcholinesterase (AChE) of Liposcelis bostrychophila and Liposcelis entomophila of paraoxon and carbosulfan were compared. The results showed that L. entomophila exhibited significantly greater specific activity of CarE than L. bostrychophila [0.045 vs. 0.012 μmol of α‐naphthyl acetate (α‐NA) hydrolysed/mg protein/min]. Moreover, L. entomophila showed higher affinity (i.e. lower K_m value) to the substrate α‐NA than L. bostrychophila (0.29 vs. 0.67 mm ). For AChE‐specific activity and affinity, no significant differences between the two species were observed. Based on the I₅₀ values, AChE of L. bostrychophila was more sensitive to paraoxon and carbosulfan than that of L. entomophila. According to inhibition kinetics, the results revealed that AChE of L. bostrychophila was 3.8‐fold more sensitive to inhibition by paraoxon than that of L. entomophila, but L. entomophila was 1.5‐fold more susceptible to carbosulfan.     

嗜卷书虱抗气调品系的选育及其适合度研究

于恒温条件下以人工饲料饲养的嗜卷书虱Liposcelis bostrychophila,在35% CO₂和1% O₂组配的气调环境中以50%左右的选择压力处理,以选育其抗气调性。至30代获得抗性品系(CA-R),抗性倍数达5.6倍,且有继续增强的基因潜能。该抗性品系对气调的抗性不太稳定,在无选择压力的情况下经室内5代饲养,抗性衰退了63.2%。CA-R品系与敏感品系在无气调选择压力的条件下相比具有一定程度的繁殖不利性;但没有发现CA-R品系在生长发育特性上的不利性。以净增殖率来衡量,CA-R品系相对于敏感品系具有0.39的适合度。     

两种昆虫生长调节剂对嗜虫书虱的致死作用

系统测定了灭幼宝（Pyriproxyfen)和烯虫酯（Methoprene) 2种昆虫生长调节剂对储藏物主要害虫嗜虫书虱（Liposcelis entomophila)的生物活性。采用饲料混药法,针对不同浓度的灭幼宝和烯虫酯分别对嗜虫书虱若虫的致死作用,对若虫发育历期,成虫繁殖力和卵孵化率的影响设置不同处理和重复。结果表明,灭幼宝对嗜虫书虱若虫的致死作用非常明显,其效果比烯虫酯好;不同浓度的灭幼宝使若虫的发育历期明显延长; 灭幼宝和烯虫酯对嗜虫书虱的成虫无杀伤作用,但对成虫的产卵量有明显的抑制作用,两者的作用浓度却相差5倍。灭幼宝对卵的孵化率影响更大,因而具有更显著的杀卵效果。     

Influence of wolbachia infection on the fitness of the stored-product pest Liposcelis tricolor (Psocoptera: Liposcelididae)

Wolbachia are maternally inherited intracellular bacteria (Rickettsiaceae) that infect a wide range of arthropods and nematodes and that are associated with various reproductive abnormalities in their hosts. In this study, the effects of removal of Wolbachia infection on development, survival, and reproduction of Liposcelis tricolor Badonnel (Psocoptera: Liposcelididae) were investigated in laboratory. The Wolbachia-free strain was obtained by the removal of Wolbachia infection by using 1% rifampicin treatment on the Wolbachia-infected strain (control) for 4 wk, and no Wolbachia gene product was detected in this strain throughout the experiment. The results showed that the removal of Wolbachia infection had negative effects on the fitness of L. tricolor. Compared with the control strain, the Wolbachia-free strain (both in the first [F1] and second [F2] generation) had prolonged developmental times, reduced survivorship of immature stages, and reduced fecundity and longevity, resulting in much smaller rm values. Using rm values, the fitness for Wolbachia-free F1 and F2 relative to the control were calculated as 0.45 and 0.27, respectively. The results of this study further confirmed our previous conclusion that Wolbachia infection have positive effects on fecundity and fertility of L. tricolor, and for optimal reproduction of L. tricolor, Wolbachia must be present in psocids.     

Rickettsia felis Infection in a Common Household Insect Pest,Liposcelis bostrychophila (Psocoptera: Liposcelidae)

Many species of Rickettsia are well-known mammalian pathogens transmitted by blood-feeding arthropods. However, molecular surveys are continually uncovering novel Rickettsia species, often in unexpected hosts, including many arthropods that do not feed on blood. This study reports a systematic molecular characterization of a Rickettsia infecting the psocid Liposcelis bostrychophila (Psocoptera: Liposcelidae), a common and cosmopolitan household pest. Surprisingly, the psocid Rickettsia is shown to be Rickettsia felis, a human pathogen transmitted by fleas that causes serious morbidity and occasional mortality. The plasmid from the psocid R. felis was sequenced and was found to be virtually identical to the one in R. felis from fleas. As Liposcelis insects are often intimately associated with humans and other vertebrates, it is speculated that they acquired R. felis from fleas. Whether the R. felis in psocids causes disease in vertebrates is not known and warrants further study.Many species of Rickettsia are well-known mammalian pathogens that are transmitted by blood-feeding arthropods via bites or feces and can cause mild to fatal diseases in humans (33). Some species are also considered potential bioterrorism agents (4). Most Rickettsia research has focused on pathogens that are found in two closely related species groups, the typhus and spotted fever groups, such as Rickettsia prowazekii, Rickettsia rickettsii, and Rickettsia typhi, the causal agents of epidemic typhus, Rocky Mountain spotted fever, and murine typhus, respectively (3, 4, 33). However, recent surveys suggest that Rickettsia bacteria are much more widespread than previously suspected and that they are being detected in novel hosts, the vast majority of which are arthropods, including many that do not feed on blood (29, 45).The number of new rickettsial species that cause diseases in humans is rapidly increasing (33). One such species that has been generating much interest in recent years is Rickettsia felis, the causative agent of a murine typhus-like disease (1, 2, 13, 16, 17, 28, 44). The disease is often unrecognized, and even though it is considered clinically mild, it can cause severe illness and death in older patients and in cases of delayed diagnosis (2). R. felis was identified only in 1990 (1) and has since been found worldwide in fleas, where it is maintained transovarially and can reach high infection rates (e.g., 86% to 94% in cat fleas) (2, 3, 44), as well as in ticks and mites (34). While experimental infections have confirmed that R. felis is transmitted to vertebrate hosts via blood feeding and that R. felis occurs in an infectious extracellular state (39), it is not known whether transmission can also occur through contamination of broken skin by infected vector feces, as in R. typhi (3, 34).A number of features distinguish R. felis from species in both the typhus and spotted fever groups. Lately, it has been proposed that R. felis be in its own group, allied with Rickettsia akari and Rickettsia australis, the causal agents of rickettsial pox and Queensland tick typhus, respectively, and a number of recently discovered strains infecting insects that do not feed on blood (16, 17, 29, 45). Moreover, R. felis was the first Rickettsia species shown to have a plasmid (28). While plasmids now appear to be quite widespread in the genus, the R. felis plasmid stands out with respect to its relatively large size and distinctive gene content (5, 6, 9, 14, 17).This study reports that a common and cosmopolitan insect, the psocid Liposcelis bostrychophila (Psocoptera: Liposcelidae) harbors R. felis. Liposcelids are the closest free-living relatives of parasitic lice (19) and are well-known for their close proximity to humans, particularly as pests in houses and grain storage facilities (8, 41). Through 16S rRNA gene sequencing, L. bostrychophila was recently shown to harbor a strain of Rickettsia (29, 30, 42). A systematic molecular characterization of this Rickettsia was conducted, demonstrating that it is authentic R. felis. Furthermore, the psocid symbiont plasmid was sequenced and was shown to be virtually identical to the plasmid from R. felis that infects cat fleas.     

Survival and reproduction of Liposcelis paeta (Pearman) (Psocoptera: Liposcelididae) when irradiated by electron beams

Electron beam irradiation has been evaluated for the survival and reproduction of Liposcelis paeta Pearman (Psocoptera: Liposcelididae). All effects increased with increasing doses from 50 to 1000 Gy when the electron energy was 2 MeV. Eggs were unable to develop into adults following a dose of 100 Gy. Complete (100%) mortality was achieved about 5 weeks after adult emergence from nymphs irradiated at 300 Gy, whereas 100% mortality of adults was achieved 9 weeks after irradiation at 300 Gy. The fecundity was greatly reduced to 1.82% of that of control when L. paeta adults were irradiated at 250 Gy. A dose of 300 Gy caused complete reproductive sterility in adults. These results suggest that the most susceptible stage was the egg stage, then nymph and adult stages. A dose of 300 Gy is suggested for quarantine treatment of commodities infested by L. paeta.     

四种杀虫剂对两种书虱羧酸酯酶和乙酰胆碱酯酶的抑制作用

选用有机磷类杀虫剂(敌敌畏、毒死蜱、对氧磷)和氨基甲酸酯类杀虫剂(丁硫克百威),通过生物测定(药膜法)和生化测定(比色法)比较了嗜卷书虱和嗜虫书虱对所选药剂的敏感差异性。根据LC50可知嗜虫书虱对所选药剂比嗜卷书虱敏感。离体酶活性分析结果显示嗜卷书虱和嗜虫书虱的羧酸酯酶只对敌敌畏敏感,且嗜卷书虱比嗜虫书虱更敏感;4种药剂对乙酰胆碱酯酶均有强烈的抑制作用,同样是嗜卷书虱比嗜虫书虱敏感。乙酰胆碱酯酶的动力学研究结果和离体酶活性测定相一致。聚丙烯酰胺凝胶电泳分析显示,4种杀虫剂离体条件下对2种书虱的酯酶同工酶的抑制能力有明显差异,其中敌敌畏的抑制力最强;但对不同同功酶的抑制趋势(对大分子的抑制似乎较强)是一致的。酶的敏感性分析结果与生测结果比较表明,2种书虱的耐药力差异与其乙酰胆碱酯酶和酯酶对药剂的敏感性无关。如要弄清耐药力机制,需做进一步研究。