Remating inhibition in female Queensland fruit flies: effects and correlates of sperm storage

Reproductive success of male insects commonly hinges both on their ability to secure copulations with many mates and also on their ability to inseminate and inhibit subsequent sexual receptivity of their mates to rival males. We here present the first investigation of sperm storage in Queensland fruit flies (Tephritidae: Bactrocera tryoni; a.k.a. 'Q-flies') and address the question of whether remating inhibition in females is directly influenced by or correlated with number of sperm stored from their first mates. We used irradiation to disrupt spermatogenesis and thereby experimentally reduce the number of sperm stored by some male's mates while leaving other aspects of male sexual performance (mating probability, latency until copulating, copula duration) unaffected. Females that mated with irradiated rather than normal males were less likely to store any sperm at all (50% vs. 89%) and, if some sperm were stored, the number was greatly reduced (median 11 vs. 120). Despite the considerable differences in sperm storage, females mated by normal males and irradiated males were similarly likely to remate at the next opportunity, indicating (1) number of sperm stored does not directly drive female remating inhibition and (2) factors actually responsible for remating inhibition are similarly expressed in normal and irradiated males. While overall levels of remating were similar for mates of normal and irradiated males, factors responsible for female remating inhibition were positively associated with presence and number of sperm stored by mates of normal but not irradiated males. We suggest seminal fluids as the most likely factor responsible for remating inhibition in female Q-flies, as these are likely to be transported in proportion to number of sperm in normal males, be uninfluenced by irradiation, and be transported without systematic relation to sperm number in irradiated males.     

Attraction and mortality of oriental fruit flies to SPLAT‐MAT‐methyl eugenol with spinosad†

Studies were conducted in 2007 and 2008 in Hawaii, USA to quantify attraction and feeding responses resulting in mortality of the male oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), to a novel male annihilation treatment (MAT) formulation consisting of specialized pheromone and lure application technology (SPLAT) in combination with methyl eugenol (ME) and spinosad (=SPLAT‐MAT‐ME with spinosad) in comparison with Min‐U‐Gel‐ME with naled (Dibrom). Our approach involved a novel behavioral methodology for evaluation of slow‐acting reduced‐risk insecticides. Methyl eugenol treatments were weathered for 1, 2, 4, and 8 weeks in California, USA, and shipped to Hawaii for bioassays. In field tests involving bucket traps to attract and capture wild males, and in toxicity studies conducted in 1 m³ cages using released males of controlled ages, SPLAT‐MAT‐ME with spinosad performed similar to or outperformed the standard formulation of Min‐U‐Gel‐ME with naled for material aged for up to 8 weeks in the 2008 tests. In laboratory feeding tests in which individual males were exposed for 5 min to the different ME treatments, mortality induced by SPLAT‐MAT‐ME with spinosad recorded at 24 h did not differ from that caused by Min‐U‐Gel ME with naled at 1, 2, and 4 weeks. Spinosad has low contact toxicity, and when mixed with SPLAT offers a reduced‐risk alternative for control of B. dorsalis, without many of the negative effects to humans and non‐targets of broad‐spectrum contact poisons such as naled. Our results indicate that SPLAT‐MAT‐ME with spinosad offers potential for control of males in an area‐wide integrated pest management (IPM) system without the need for conventional organophosphates.     

桔小实蝇对五种芒果气味挥发性物质的行为反应

为了探究寄主气味挥发物对桔小实蝇行为的影响,本研究应用Y型嗅觉仪测定了桔小实蝇Bactrocera dorsalis(Hendel)成虫对5种芒果气味挥发性物质(乙酸乙酯,α-蒎稀,异松油烯,3-蒈烯和石竹烯)及5种物质混合物的行为反应。结果表明,桔小实蝇雌、雄虫对5种物质及5种物质混合物的行为反应无显著性差异。结果还揭示,桔小实蝇成虫对乙酸乙酯有明显的趋性反应,对异松油烯有明显的忌避反应,而对α-蒎烯、3-蒈烯、石竹烯和5种物质的混合物均没有明显的行为定向反应。说明桔小实蝇成虫对芒果不同气味挥发物的行为反应存在差别,这可为研发桔小实蝇成虫防治新方法提供基础信息。     

Manipulation of insect gut microbiota towards the improvement of Bactrocera oleae artificial rearing

Bactrocera oleae (Rossi) (Diptera: Tephritidae) is the main pest of olive trees (Olea europaea L.), causing major damages in olive crops. Improvement of mass rearing is a prerequisite for the successful development of large-scale sterile insect technique (SIT) applications. This can be achieved through the enrichment of artificial diets with gut bacteria isolates. We assessed the efficiency of three gut bacteria previously isolated from Ceratitis capitata (Wiedemann), and four isolated from B. oleae, as larval diet additives in both live and inactivated/dead forms. Our results showed that dead Enterobacter sp. AA26 increased pupal weight, whereas both live and dead cells increased pupal and adult production and reduced immature developmental time, indicating that its bacterial cells serve as a direct nutrient source. Live Providencia sp. AA31 improved pupal and adult production, enhanced male survival under stress conditions, and delayed immature development. Dead Providencia sp. AA31, however, did not affect production rates, indicating that live bacteria can colonize the insect gut and biosynthesize nutrients essential for larval development. Live and dead Bacillus sp. 139 increased pupal weight, accelerated immature development, and increased adult survival under stress. Moreover, live Bacillus sp. 139 improved adult production, indicating that Bacillus cells are a direct source of nutrients. Dead Serratia sp. 49 increased pupal and adult production and decreased male survival under stress conditions whereas live cells decreased insect production, indicating that the live strain is entomopathogenic, but its dead cells can be utilized as nutrient source. Klebsiella oxytoca, Enterobacter sp. 23, and Providencia sp. 22 decreased pupal and subsequent adult production and were harmful for B. oleae. Our findings indicate that dead Enterobacter sp. AA26 is the most promising bacterial isolate for the improvement of B. oleae mass rearing in support of future SIT or related population suppression programs.     

短时高温胁迫对瓜实蝇生长发育及繁殖的影响

为明确瓜实蝇对短时高温胁迫的耐受性。利用人工气候箱模拟短时高温胁迫,测定了不同高温处理(34、36、38、40、42、44、46、48℃)12 h,对不同发育阶段瓜实蝇的存活率和生长发育的影响。结果表明短时高温显著影响瓜实蝇的存活,随温度升高,瓜实蝇各虫态的存活率逐渐降低;高温处理12 h后瓜实蝇卵、幼虫、蛹、雌成虫、雄成虫的致死中温度LT 50分别为35.48、37.55、41.85、43.62、43.32℃;34~42℃短时高温胁迫对瓜实蝇各虫态的发育历期无明显影响,44℃时其发育历期均显著增长;46℃、48℃处理下各虫态死亡率较高,不能正常发育;随着处理温度的升高,雌成虫产卵前期不断增长,单雌产卵量呈下降趋势,成虫寿命不断缩短,后代雌性比增大。44℃及以上的短时高温胁迫不利于瓜实蝇的生长发育,40℃及以上的短时高温胁迫不利于瓜实蝇的繁殖,雌性瓜实蝇对短时高温的胁迫的适应性强于雄性,随着处理温度的升高,后代雌性比例增大。     

Yeast hydrolysate supplement increases starvation vulnerability of Queensland fruit fly

Post‐teneral diets containing yeast hydrolysate are reported to increase longevity, reproductive development and sexual performance of Queensland fruit fly (‘Q‐fly’) Bactrocera tryoni Froggatt (Diptera: Tephritidae). Consequently, diets including yeast hydrolysate are recommended for sterile Q‐flies before release in sterile insect technique (SIT) programmes. However, in some tephritids, diets including yeast hydrolysate are associated with an increased vulnerability to starvation. In the present study, the effects of yeast hydrolysate supplementation before release are considered with respect to the longevity of released Q‐fly when food becomes scarce. Experiments are carried out in three settings of varying resemblance to field conditions: 5‐L laboratory cages, 107‐L outdoor cages and 14 140‐L field cages containing potted citrus trees. In all experimental settings, compared with flies that received only sucrose, male and female Q‐flies that are provided with yeast hydrolysate during the first 2 days of adult life have a significantly shorter survival when subsequently deprived of food. Yeast supplementation appears to commit Q‐flies to a developmental trajectory that renders them more vulnerable to starvation. The practical significance of these findings for SIT depends on how often the releases are carried out under conditions in which Q‐flies experience extreme food shortages in the field.     

Field evaluation of the relative attractiveness of enriched ginger root oil (EGO)lure and trimedlure for African Ceratitis species (Diptera: Tephritidae)

The males of some fruit flies (Diptera: Tephritidae) are known to be attracted to specific parapheromones. The trapping results between trimedlure (TML) and enriched ginger root oil (EGOlure) were compared at two experimental sites in Morogoro (Central Tanzania) for a period of 12 weeks co‐inciding with the main citrus season. Both attractants captured a comparable diversity of fruit flies, except that EGOlure also attracted fruit flies, such as Ceratitis cosyra, not normally found in TML‐baited traps. Both EGOlure and TML attracted mainly or exclusively male fruit flies, but the catches with EGOlure were equal or superior to those with TML. It is concluded that EGOlure should be considered as a suitable alternative for TML in detection, monitoring and control programs for African fruit flies of the genus Ceratitis. It has the added advantage that it combines the attractiveness with regard to species spectrum of both TML and terpinyl acetate.     

应用寄生蜂和不育雄虫防控田间橘小实蝇

食品安全已成为当今社会广受关注的问题。尽管国内的一些学者就阿里山潜蝇茧蜂[Fopius arisanus(Sonan)]和橘小实蝇SIT技术分别开展了不同程度的研究,然而关于应用该蜂和不育雄虫对橘小实蝇[Bactrocera dorsalis(Hendel)]进行田间联合防治的研究还未见相关报道。为此,为探讨天敌昆虫和雄性不育技术对橘小实蝇的田间综合治理(IPM)技术,开展了阿里山潜蝇茧蜂和橘小实蝇不育雄虫的田间释放试验:首先应用性引诱剂降低田间野生雄虫数量;而后,释放橘小实蝇不育雄虫,进一步降低田间橘小实蝇种群数量;最后,释放阿里山潜蝇茧蜂对田间残余的橘小实蝇卵进行追踪寄生,以达到持续控制的目的。通过监测整个防控期田间橘小实蝇的种群数量以及果实受害率和防治效果作为评价指标评价该防治方法的综合防治效果。结果表明:应用阿里山潜蝇茧蜂、橘小实蝇不育雄虫并配合引诱剂的综合措施对橘小实蝇的种群数量有明显控制作用,田间总体防治效果可达90%左右。此外,提出完善橘小实蝇田间防控措施的建议,并提出该研究的不足以及有待进一步开展研究的方向。     

Purification and biochemical characterization of glutathione S-transferases from four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Glutathione S-transferases (GSTs) are a group of detoxification enzymes that catalyze the nucleophilic addition of glutathione to a wide variety of endogenous and xenobiotic compounds. In this study, GSTs were purified from four field populations of Bactrocera dorsalis with different insecticide susceptibilities by glutathione-agarose affinity chromatography. The populations were collected from Dongguan (DG) and Guangzhou (GZ) of the Guangdong Province, Haikou of the Hainan province (HN), and Kunming of the Yunnan province (YN), China. Differences in GST characteristics among the four populations were studied using purified enzyme samples through comparative SDS-PAGE, kinetic, and inhibition experiments. The specific activities of the purified enzymes were similar, but the purification yield of the GZ population (31.54%) was the lowest. SDS-PAGE analysis showed only one band at approximately 23 kDa for these four populations. Kinetic analyses showed that the affinities of the purified GSTs from the GZ and YN populations for 1-chloro-2.4-dinitrobenzene (CDNB) were much higher than those of GSTs from the other two populations, whereas the HN population had the highest catalytic capability in terms of V(max) value. The optimum temperature for CDNB conjugation was 37 °C and the optimum pH was 7.5 in all four populations. Inhibition kinetics showed that ethacrynic acid, diethyl maleate, tetraethylthiuram disulfide, curcumin, bromosulfalein, and β-cypermethrin had excellent inhibitory effects on GSTs in the four populations of B. dorsalis, but the low inhibitory effects of malathion and avermectin did not differ between populations. These results suggest that GSTs may have a role in detoxification of β-cypermethrin in B. dorsalis.     

Dietary wheat germ oil influences gene expression in larvae and eggs of the oriental fruit fly

Changes in animal nutrition, particularly essential dietary components, alter global gene expression patterns. Our goal is to identify molecular markers that serve as early indicators of the quality of insect culture media. Markers of deficient culture media will increase the efficiency of developing optimal systems for mass rearing beneficial insects and some pest species because decisions on culture media quality can be made without waiting through one or several life cycles. The objective of our current study is to discover molecular markers of essential dietary lipid deficiency in the oriental fruit fly, Bactrocera dorsalis. We reared groups of fruit flies separately on media either devoid of or supplemented with wheat germ oil (WGO) and analyzed gene expression in third instar larvae and F(1) eggs using 2D electrophoresis. Gel densitometry revealed significant changes in expression levels of genes encoding eight proteins in larvae and 22 proteins in eggs. We identified these proteins by using mass spectrometry (MALDI TOF/TOF) and bioinformatic analyses of the protein sequences. Among these, we identified one gene encoding the receptor of activated C Kinase 1 (RACK1) that increased in expression by 6.8-fold in eggs from adults that were reared as larvae on media supplemented with WGO. RACK1 is an essential component of at least three intracellular signal transduction pathways, making it a good molecular marker candidate of lipid deficiency in fruit flies and possibly many other insect species.