Nasonia vitripennis venom causes targeted gene expression changes in its fly host

Parasitoid wasps are diverse and ecologically important insects that use venom to modify their host's metabolism for the benefit of the parasitoid's offspring. Thus, the effects of venom can be considered an ‘extended phenotype’ of the wasp. The model parasitoid wasp Nasonia vitripennis has approximately 100 venom proteins, 23 of which do not have sequence similarity to known proteins. Envenomation by N. vitripennis has previously been shown to induce developmental arrest, selective apoptosis and alterations in lipid metabolism in flesh fly hosts. However, the full effects of Nasonia venom are still largely unknown. In this study, we used high throughput RNA sequencing (RNA‐Seq) to characterize global changes in Sarcophaga bullata (Diptera) gene expression in response to envenomation by N. vitripennis. Surprisingly, we show that Nasonia venom targets a small subset of S. bullata loci, with ~2% genes being differentially expressed in response to envenomation. Strong upregulation of enhancer of split complex genes provides a potential molecular mechanism that could explain the observed neural cell death and developmental arrest in envenomated hosts. Significant increases in antimicrobial peptides and their corresponding regulatory genes provide evidence that venom could be selectively activating certain immune responses of the hosts. Further, we found differential expression of genes in several metabolic pathways, including glycolysis and gluconeogenesis that may be responsible for the decrease in pyruvate levels found in envenomated hosts. The targeting of Nasonia venom effects to a specific and limited set of genes provides insight into the interaction between the ectoparasitoid wasp and its host.     

Effect of exposure time on mass‐rearing production of the olive fruit fly parasitoid,Psyttalia lounsburyi (Hymenoptera: Braconidae)

Classical biological control programmes rely on mass production of high‐quality beneficial insects for subsequent releases into the field. Psyttalia lounsburyi (Silvestri) (Hymenoptera: Braconidae) is a koinobiont larval–pupal endoparasitoid of tephritid flies that is being reared to support a classical biological control programme for olive fruit fly in California. The mass‐rearing system for a P. lounsburyi colony, initiated with insects originally collected in Kenya, was evaluated with the goal of increasing production, while at the same time reducing time requirements for rearing in a quarantine facility. We tested the effect of exposure time of a factitious host Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), on parasitization, adult production, superparasitism, and sex ratio of P. lounsburyi and survival of the host. Parasitization rates were highest (31%) at 3‐ and 4‐hr exposure times, while adult production (i.e., emergence of wasp progeny) was highest (16%) at the 2‐hr exposure time. Superparasitism over the course of the study was 1.5% and did not appear to be a factor affecting parasitoid production. The sex ratio of wasp progeny was male‐biased and did not vary significantly over different exposure times. The rate of stings on host larvae increased with exposure time and was consistent with decreases in pupal eclosion from larvae and emergence rate of adult flies. When compared to current rearing procedures, the 2‐hr exposure time resulted in an overall 2.8‐fold increase in P. lounsburyi production when standardized for time.     

Development of Sarcophaga dux (diptera: Sarcophagidae) at constant temperatures and differential gene expression for age estimation of the pupae

Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly species with potential forensic value for estimating minimum postmortem interval (PMI_min). The basic developmental data and precise age estimates of the pupae are significant for PMI_min estimation in forensic investigations. In the present study, we investigated the development data of that species at seven constant temperatures varying from 16 °C to 34 °C, including body length changes of the larve, developmental duration and accumulated degree hours of the preadults. Several reference genes for relative quantification of the differentially expressed genes (DEGs) were firstly selected and evaluated in the pupae of different ages under different temperatures. The DEGs of the insects during the pupal period at different constant temperatures (34, 25 and 16 °C) were further analyzed for more precise age estimation. The results showed that the developmental durations of the preadults at 16, 19, 22, 25, 28, 31 and 34 °C were 1478.6 ± 18.3 h, 726.1 ± 15.8 h, 538.5 ± 0.9 h, 394.1 ± 9.5 h, 375.6 ± 10.8 h, 284.1 ± 7.3 h, and 252.5 ± 6.1 h, respectively. The developmental threshold temperature the flies was 12.27 ± 0.35 °C, and the thermal summation constant was 5341.71 ± 249.29° hours. The most reliable reference genes during the pupal period at different temperatures were found: GST1 and 18S rRNA for the 34 °C group, GST1 and RPL49 for 25 °C, and 18S rRNA and 28S rRNA for 16 °C. The four differential expression genes (Hsp60, A-alpha, ARP, and RPL8) have the potential to be used for more precise age estimation of pupal S. dux. This work provides important basic developmental data and a more precise age estimation method for pupal S. dux, and improves the value of this species for PMI_min estimation in forensic investigations.     

Plasticity in life features,parasitism and super-parasitism behavior of Bracon hebetor,an important natural enemy of Galleria mellonella and other lepidopteran host species

The wasp, Bracon hebetor Say, is an important potential biocontrol agent of a wide range of lepidopteran insect species. The current study was subjected to compare these ectoparasitoid fitness traits on different host species belonging to the order Lepidoptera. Moreover, we determine the population dynamic with different host densities, sex ratio patterns, super-parasitism, longevity, paralysis success, and reproductive potential under laboratory conditions. Our results revealed that oviposition increased with an increase in host density, while the adult emergence and egg hatching were decreased due to the super-parasitism on host larvae. A higher male and female-biased population were observed when virgin and mated females offered fresh hosts. Adults' longevity was recorded more in females than males when kept only with bee honey + royal jelly + host larvae. The mean duration of egg-adult development was recorded higher on Galleria mellonella and lowest on Busseola fusca. The parasitization rate, super-parasitism, and cumulative fecundity of mated and virgin female wasps with different host species were observed higher on specific host Galleria mellonella while it was lower on Phthorimaea operculella. Furthermore, the parasitoids having mating experience preferred fresh, while the parasitoids' lack of mating experiences preferred paralyzed host under olfactometer test. Besides, this research has produced novel facts on the biology of parasitic wasp, B. hebetor that may guide the advancement of sustainable biological control programs to control lepidopteran pests.     

Effects of two insect growth regulators and a neonicotinoid on various life stages of <Emphasis Type="Italic">Aphytis melinus</Emphasis> (Hymenoptera: Aphelinidae)

The effect of two insect growth regulators and a neonicotinoid insecticide were tested on immature stages and adults of the parasitoid Aphtyis melinus DeBach (Hymenoptera: Aphelinidae), a key natural enemy of California red scale, Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae), in California. No significant effects of the insect growth regulators on survival or development to the adult stage were found when the parasitoid was treated at any of the egg, larval, or pupal stages. The broad-spectrum neonicotinoid acetamiprid also showed no significant effect on the development of A. melinus to the pupal stage, probably because immature stages of this ectoparasitoid are protected under the cover of its armored scale host. However, 48 h exposure of adults to acetamiprid residues following emergence resulted in high levels of wasp mortality. Aphytis melinus adults treated with either of the two insect growth regulators as larvae survived 48 h exposure to pesticide residues as adults and showed levels of fecundity comparable with control insects. We conclude that the two insect growth regulators are compatible with augmentative releases of A. melinus but that treatments of acetamiprid should be avoided in situations where biological control by this parasitoid is important.     

Bionomic notes on Pachysomoides sp. (Hymenoptera: Ichneumonidae), a parasitoid of the Neotropical social wasp Polistes satan Bequaert (Hymenoptera: Vespidae)

Although most polistine wasp species are found in the Neotropical region, mainly in Brazil, only a very limited number of South American parasitoids or parasites are known to exist. We assessed the frequency of a hymenopterous parasitoid, Pachysomoides sp. (Ichneumonidae, Cryptinae), in the nests of the Brazilian independent‐founding wasp Polistes satan and compared the rates of the parasitization of P. satan by Pachysomoides sp. between the dry (winter) and wet (summer) seasons. Pachysomoides sp. larvae were seen to feed on P. satan pupa and were found in both the upper and lower parts of the host pupal cell (ca. 10 individuals in each host pupal cell). Approximately one‐third of the pupal cells in the P. satan colonies were parasitized in the dry season, whereas there were no parasitized pupal cells in the wet season. Consequently, the rates of parasitization by Pachysomoides sp. were significantly greater during the dry season than during the wet season due to unknown reasons.     

蝶蛹金小蜂寄生对菜粉蝶体液免疫反应的影响(英文)

以蝶蛹金小蜂及其寄主菜粉蝶为研究对象 ,研究了内寄生蜂对寄主体液免疫反应的影响。当寄主蛹被寄生后 1 2h或第 4和 5d时 ,血淋巴中酚氧化酶活性明显增高。寄生蛹血淋巴中血细胞凝集素活性始终高于针刺和未寄生蛹 ;同样 ,寄生蛹血淋巴的抗菌活性也明显增强 ,而后两者处理蛹的活性则很微弱。由此可知 ,该蜂寄生能引起寄主体液免疫因子活性的不同程度的变化     

Limited protection of the parasitoid Pachycrepoideus vindemiae from Drosophila suzukii host‐directed spinosad suppression

Laboratory trials were conducted to determine whether the spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), puparium can provide an effective physical barrier to protect immature stages of the pupal parasitoid Pachycrepoideus vindemiae (Rondani) (Hymenoptera: Pteromalidae) from spinosad treatments. Spinosad insecticides are currently an important suppression strategy for D. suzukii in organically managed fruit orchards although they are well known to cause mortality in hymenopteran parasitoids. High adult P. vindemiae female mortality (83%) occurred within 24 h of exposure to D. suzukii pupae treated with 10 mg a.i. l^?1 spinosad and female parasitoids did not avoid the pupae treated with similar low levels of spinosad in choice tests that included untreated pupae. Pachycrepoideus vindemiae develops as an idiobiont ectoparasitoid on host fly pupa within the sclerotized host puparium. Significant P. vindemiae survival and emergence was recorded when parasitized D. suzukii puparia were exposed to field treatment levels of spinosad; however, the parasitoid survival was dependent on the time of the spinosad treatment of the host post‐parasitization. Significant parasitoid survival occurred when the host puparia were treated at 2 weeks when the parasitoid was in the pupal stage but did not occur when the host puparia were treated at 1 week post‐parasitization, when the parasitoids were still in a larval stage. The parasitoid adults consumed or otherwise came in contact with residual degrading spinosad when they exited the treated host, and consequently high and low adult parasitoid mortality occurred when the adults emerged from puparia treated at 2 and 1 week(s), respectively. Our study indicates that generally the integration of P. vindemiae parasitism into a sustainable D. suzukii management program is not compatible with spinosad treatments, although P. vindemiae in the pupal stage inside sclerotized host puparia appear to be minimally impacted by spinosad treatments, provided that the spinosad degrades before parasitoid emergence.     

EFFECT OF PARASITIZATION BY THE PUPAL ENDOPARISITOID,PTEROMALUS PUPARUM (HYMENOPTERA: PTEROMALIDAE) ON HUMORAL IMMUNE REACTIONS OF PIERIS RAPAE (LEPIDOPETERA: PIERIDAE)*

Abstract Studies on the effect of parasitization by the endoparasitoid on host humoral immune reactions are carried out with the pupal endoparisitic wasp, Pteromalus puparum, and its host, Pieris rapae. Phenoloxidase (PO) activity of parasitized hosts hemolymph increased significantly at 12 h, day four and day five after parasitization. Hem‐agglutination activity of parasitized hosts hemolymph was always higher than that of wounded and unparasitized ones. Moreover, antibacterial activity of parasitized hosts hemolymph became more and more stronger, whilst wounded and unparasitized pupae only owned a weak antibacterial activity. It suggested that activities of humoral immune factors of Pieris rapae could be influenced to some degrees by P. puparum.     

Host acceptance behavior of the Japanese aquatic wasp,Agriotypus gracilis (Hymenoptera: Ichneumonidae) toward the caddisfly host,Goera japonica (Trichoptera: Limnephilidae)

The host acceptance behavior of the Japanese aquatic wasp,Agriotypus gracilis Waterston, an ectoparasitoid of the sand case building caddisfly,Goera japonica Banks was investigated in the laboratory. Female wasps were observed to enter the water by walking down a stone protruding from the water surface. Antennae were held backward and not utilized in searching for hosts under the water. Female wasps examined hosts from the outside of their cases by 2 consecutive steps, “turning” and “probing”. Turning behavior, in which female wasps move between the anterior and posterior ends of host cases, may be related to the measurement of case size. Host stages are considered to be discriminated by probing, in which females probe host cases with their sheathing ovipositors. Female wasps most frequently accepted and oviposited on pupal and prepupal hosts.     

Fate of polydnavirus DNA of the egg-larval parasitoid Chelonus inanitus in the host Spodoptera littoralis

In situ hybridizations show that 5 min after parasitization, polydnavirus DNA is in close vicinity of the parasitoid egg, but 5 h later also in the yolk and partially in the host embryo. Fifteen hours after parasitization, the viral DNA is seen all over the host embryo and hardly in the yolk. The tissue distribution of the viral DNA was analysed and quantified by dot blots in the fifth instar parasitized larvae. On a per host basis, haemocytes and fat body contained the highest amount of viral DNA, while nervous tissue, intestinal tract and carcass contained less. Of the three viral segments tested, all were found in all tissues. Relative to the quantity of host DNA, viral DNA was most abundant in haemocytes, about five times less abundant in fat body and nervous tissue and about 25 times less abundant in intestinal tract. The total quantity of viral DNA per host was 444+/-145 pg which is similar to the quantity injected by the wasp; thus, the viral DNA persists throughout parasitization. The parasitoid larva contains 820+/-80 pg viral DNA integrated in the genome. This illustrates that the dose of viral DNA injected in virions represents approximately one third of the total viral genomic information present in a host at a late stage of parasitism.     

Cold hardiness of the fly pupal parasitoid Nasonia vitripennis is enhanced by its host Sarcophaga crassipalpis

Supercooling points (SCPs) and low temperature survival were determined for diapausing and nondiapausing larvae of the ectoparasitoid Nasonia vitripennis. Neither nondiapausing nor diapausing larvae could survive tissue freezing. The SCP profiles were nearly identical for nondiapause-destined (-27 degrees C) and diapausing larvae (-25 degrees C), but these values were not indicative of the lower limits of tolerance in either type of larvae: larvae were killed by chilling at temperatures well above the SCP. Diapausing larvae could withstand low temperature exposures 3-8 times longer than their nondiapausing counterparts. Low temperature survival was enhanced in diapausing and nondiapausing larvae by their encasement within the puparium of the host flesh fly, SARCOPHAGA CRASSIPALPIS: the LT(50)s determined for nondiapausing and diapausing larvae enclosed by fly puparia were 2-3 times higher than values calculated for larvae removed from the puparia. Additional low temperature protection was gained through acquisition of host cryoprotectants during larval feeding: nondiapausing parasitoid larvae that fed on diapausing flesh fly pupae with high levels of glycerol were able to survive exposure to a subzero temperature 4-9 times longer than wasps reared on nondiapausing fly pupae that contained lower quantities of glycerol. Alanine may also contribute to the cold hardiness of N. vitripennis, as evidenced by the fact that larvae feeding on diapausing fly pupae both contained higher concentrations of alanine and exhibited greater cold hardiness. The results thus demonstrate that several critical features of cold hardiness in the wasp are derived from biochemical and physical attributes of the host.     

枸杞木虱啮小蜂寄生行为及生物学特性的研究

枸杞木虱啮小蜂Tetrastichussp .是枸杞木虱ParatriozasinicaYang&Li若虫的外寄生蜂 ,自然寄生率可达 86 5 %。在温度为 2 0～ 2 5℃的室内条件下 ,完成 1个世代需 1 4～ 2 0d。成虫寿命与取食有关 ,取食 1 5 %的蜂蜜水 ,雌性成虫寿命为 1 2～ 1 7d,雄性成虫为 4～ 6d。成蜂最喜欢在 3龄和 4龄若虫上产卵。自然种群消长与寄主虫口密度相关。每年 7月到 8月下旬该蜂寄生率高 ,9月份下降 ,1 0月份几乎不见。