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Transfer of methyl-branched hydrocarbons from the parasitoid, Eretmocerus mundus, to silverleaf whitefly nymphs during oviposition.

The parasitic wasp Eretmocerus mundus (Hymenoptera: Aphelinidae), a natural enemy of the silverleaf whitefly Bemisia argentifolii (Homoptera: Aleyrodidae), deposits eggs beneath nymphs and not within them. Experiments were designed to establish whether ovipositing E. mundus females leave marking chemicals on nymphs to enable searching females to discriminate parasitized from unparasitized hosts. Cuticular lipids from three experimental treatment groups were characterized: parasitoid-exposed nymphs that had a parasitoid egg between the nymph and leaf; control nymphs not exposed to E. mundus; and a third treatment condition of parasitized nymphs, held for 10 days after wasp exposure. Lipids were solvent-extracted from the nymphal cuticles of the various treatment groups and the lipid components were characterized and quantified by gas chromatography and mass spectrometry. Results indicated the presence of quantities of C31 and C33 dimethylalkanes only from parasitoid-exposed groups of nymphs and not in the extracts from control nymphs or the parasitized nymphs after 10-day exposure. Furthermore, the C31 and C33 dimethylalkanes were shown to be major lipid components of the hexane extracts from E. mundus females. Since the lipids were removed from parasitoid-exposed nymphs before interaction with hatched parasitoid larvae, the findings indicated that the dimethylalkanes were transferred onto nymphal cuticles by ovipositing E. mundus females.

Alkanes↗

Partial elucidation of Trichogramma putative sex pheromone at trace levels by solid-phase microextraction and gas chromatography-mass spectrometry studies.

Virgin females of the minute parasitoid wasp Trichogramma turkestanica produce about 2 pg/h of two putative sex pheromonal compounds. These compounds could be successfully sampled during 20-50 h with SPME from 1.8 mL vials, containing 50-110 wasps and analysed by GC-MS. Accurate mass measurements at the 1 ng scale were possible with an internal standard of sulphur. One compound with MW 236 was identified as a C17H32 hydrocarbon while the other compound was the corresponding allylic alcohol with composition C17H32O. The alcohol could be silylated on-fibre and its mass spectrum suggested the presence of a conjugated 2,4-diene moiety. A miniaturised solvent extraction system for SPME needles, using 5 microL of acetone in a microtube was developed. After reaction of the extracted volatiles with the dienophile 4-methyl-1,2,4-triazoline-3,5-dione (MTAD), GC-MS of the MTAD adducts confirmed the presence of a diene. Interpretation of the combined mass spectral data, in combination with retention indexes of both compounds on non-polar and polar columns, suggested 2,6,8,12-tetramethyltrideca-2,4-diene and 2,6,8,12-tetramethyltrideca-2,4-dien-1-ol as most probable structures. These compounds have not been described previously. Biogenetically, they are most likely polyketides made up of a C4 starter unit that has been elongated with C2 and C3 units. Further biological and synthetic studies are necessary to prove their role as sex pheromone, confirm the proposed structures and determine the correct stereochemistry of the double bonds and the methyl groups.

Animals↗

Sol i 1, the phospholipase allergen of imported fire ant venom.

BACKGROUND: Sol i 1, the venom phospholipase of imported fire ant venom is an important allergen and exhibits some cross-reactivity with IgE antibodies from patients sensitized to other Hymenoptera venoms. OBJECTIVE: To determine the primary structure of Sol i 1 and evaluate the roles of protein and carbohydrate epitopes in its cross-reactivity. METHODS: Sol i 1 was purified from venom, proteolytic peptides prepared and amino acid sequences obtained. The cDNA for Sol i 1 was cloned, sequenced, and compared with sequences of other wasp venom phospholipases. The role of carbohydrate epitopes in the cross-reactivity with other Hymenoptera venoms was studied by RAST inhibition. RESULTS: The sequence identified Sol i 1 as a lipase of the GX class, lipoprotein lipase superfamily, pancreatic lipase homologous family and RP2 subgroup phospholipases as are the vespid venom phospholipases. The 148 residues identified by amino acid sequencing represent about 48% of the translated cDNA sequence. Sol i 1 was 31-32% identical to yellow jacket phospholipases. The identical regions of sequence were clustered in the domain which forms the serine hydrolase active site. Mannosylated N-glycans could completely inhibit binding of IgE from honeybee venom sensitized patients to Sol i 1. Inhibition by glycan of IgE binding from yellow jacket venom sensitized patients was low or absent for three of eight sera and substantial, but not complete for five sera. CONCLUSIONS: Sol i 1 is related to wasp venom phospholipases. Cross-reactivity with honeybee venom is caused by carbohydrate, whereas cross-reactivity with yellow jacket venom involves reactivity with both carbohydrate determinants of hyaluronidase and high molecular weight proteins and phospholipase protein determinants.

Allergens↗

Dynamics of in vivo power output and efficiency of Nasonia asynchronous flight muscle.

By simultaneously measuring aerodynamic performance, wing kinematics, and metabolic activity, we have estimated the in vivo limits of mechanical power production and efficiency of the asynchronous flight muscle (IFM) in three species of ectoparasitoid wasps genus Nasonia (N. giraulti, N. longicornis, and N. vitripennis). The 0.6 mg animals were flown under tethered flight conditions in a flight simulator that allowed modulation of power production by employing an open-loop visual stimulation technique. At maximum locomotor capacity, flight muscles of Nasonia are capable to sustain 72.2 +/- 18.3 W kg(-1) muscle mechanical power at a chemo-mechanical conversion efficiency of approximately 9.8 +/- 0.9%. Within the working range of the locomotor system, profile power requirement for flight dominates induced power requirement suggesting that the cost to overcome wing drag places the primary limit on overall flight performance. Since inertial power is only approximately 25% of the sum of induced and profile power requirements, Nasonia spp. may not benefit from elastic energy storage during wing deceleration phases. A comparison between wing size-polymorphic males revealed that wing size reduction is accompanied by a decrease in total flight muscle volume, muscle mass-specific mechanical power production, and total flight efficiency. In animals with small wings maximum total flight efficiency is below 0.5%. The aerodynamic and power estimates reported here for Nasonia are comparable to values reported previously for the fruit fly Drosophila flying under similar experimental conditions, while muscle efficiency of the tiny wasp is more at the lower end of values published for various other insects.

Animals↗

Two coexisting lines of the endoparasitoid Venturia canescens show differences in reproductive success under conspecific superparasitism.

In a laboratory colony of the endoparasitic wasp Venturia canescens Grav. (Hymenoptera: Ichneumonidae), two genetically distinct lines (RP, RM) appear to coexist sympatrically. The two lines display pronounced differences in ovarian morphology, parasitism behaviour and number of offspring produced under competing superparasitism. Since V. canescens is a solitary endoparasitoid, larvae inside superparasitised hosts must compete for host possession. We examined the outcome of conspecific superparasitism between the wasp lines with different time intervals between ovipositions. The results showed that the competitive abilities of the two lines were not symmetrical. Further, the RM-line won a significantly higher fraction (around 60%) of the overall contests. Dissection of parasitoid larvae from their hosts indicated that most contests between competing larvae had occurred within the first 24 h of the eggs hatching, suggesting the advantage of the RM-line relates to physical combat. It was previously thought that the coexistence of the two lines was exclusively due to maternal effects. The results of this study indicate for the first time that these differences are based on phenotypic variations in both the larval offspring and the mother.

Age Factors↗

Lifetime egg maturation by host-deprived Venturia canescens.

The study investigated egg maturation and deposition by the endoparasitoid wasp Venturia canescens under conditions of host deprivation. Female V. canescens maintained without hosts began to deposit eggs onto the sides of the culture vessel on the day of eclosion. The maturation of additional eggs was not inhibited once the maximum oviduct egg load was reached but rather continued for the duration of the experiment (up to 39 days), at a rate of around 5.8% of the remaining unmatured eggs per day. Following host access, wasps matured additional eggs at an increased rate. Artificial damage to the ovipositor resulted in a reduced rate of egg maturation even though the oviducts were partly egg depleted, while damage to the auxiliary valvulae had no effect. These results suggest two conclusions. Under conditions of host deprivation, the rate at which eggs are matured is determined by the rate of synthesis of precursors by the fat body that in turn is modified by feedback from the ovipositor, induced by physical stimulation. Further, the discarding of eggs is due to the involuntary unidirectional movement of eggs down the oviduct, facilitated by the ongoing maturation of additional eggs.

Animals↗

Morphogenesis and cytopathic effects of the Diachasmimorpha longicaudata entomopoxvirus in host haemocytes.

The Diachasmimorpha longicaudata entomopoxvirus (DlEPV), the first reported symbiotic entomopoxvirus, occurs in the venom apparatus of D. longicaudata female wasps and is introduced into Anastrepha suspensa larvae during parasitism. The DlEPV 250-300 kb double stranded DNA genome encodes putative proteins having 30 to >60% amino acid identity with poxvirus homologs such as DNA helicase, DNA-dependent RNA polymerase, and the poxvirus-specific rifampicin resistance protein. Although the molecular characterization of DlEPV is progressing, little is known about its morphogenesis in and effects on host haemocytes. This paper describes (1) haemocytes of third instar A. suspensa, (2) DlEPV infection and morphogenesis, and (3) DlEPV-induced changes in haemocytes. A. suspensa third instars have 3-4 haemocyte morphotypes. Dot blots of DNA from infected haemocytes hybridized with a digoxigenin-labeled DlEPV genomic probe as early as 4 h post-parasitism (hpp) and the intensity of the signal increased with time through 40 hpp. Immunofluorescence microscopy localized DlEPV proteins in cytoplasmic (but not nuclear) sites of infected haemocytes, within 24-36 hpp. Electron microscopy confirmed the presence of viral envelopes, immature spheroids with centric nucleoids, budding virus, and extracellular enveloped virus in three haemocyte types, 24-84 hpp and later. Infected haemocytes exhibited blebbing, DNA concatenation, and inability to encapsulate sephadex beads in vitro. These data indicate that DlEPV disrupts the normal function of host haemocytes, thereby insuring the successful development of D. longicaudata offspring and as such should be regarded as a symbiont of the wasp.

Animals↗

Flowers as sensory billboards: progress towards an integrated understanding of floral advertisement.

This paper explores the landscape of integrated sensory signals that are produced by flowers, and the contextual information that modulates pollinator responses to such signals. Chiloglottis orchids are pollinated by mimicking the pheromone and posture of female thynnine wasps, but floral height provides the context within which male wasps respond to these signals. The odor and appearance of carrion attract blowflies to Helicodiceros inflorescences, but flies are more likely to enter the floral chamber when heat is present as a contextual cue. Finally, fragrance, UV-visual cues and echo fingerprints are redundant signals that Glossophaga bats can use to find flowers, depending on prior experience and the photic environment.

Animals↗

Microplitis demolitor bracovirus genome segments vary in abundance and are individually packaged in virions.

Polydnaviruses (PDVs) are distinguished by their unique association with parasitoid wasps and their segmented, double-stranded (ds) DNA genomes that are non-equimolar in abundance. Relatively little is actually known, however, about genome packaging or segment abundance of these viruses. Here, we conducted electron microscopy (EM) and real-time polymerase chain reaction (PCR) studies to characterize packaging and segment abundance of Microplitis demolitor bracovirus (MdBV). Like other PDVs, MdBV replicates in the ovaries of females where virions accumulate to form a suspension called calyx fluid. Wasps then inject a quantity of calyx fluid when ovipositing into hosts. The MdBV genome consists of 15 segments that range from 3.6 (segment A) to 34.3 kb (segment O). EM analysis indicated that MdBV virions contain a single nucleocapsid that encapsidates one circular DNA of variable size. We developed a semi-quantitative real-time PCR assay using SYBR Green I. This assay indicated that five (J, O, H, N and B) segments of the MdBV genome accounted for more than 60% of the viral DNAs in calyx fluid. Estimates of relative segment abundance using our real-time PCR assay were also very similar to DNA size distributions determined from micrographs. Analysis of parasitized Pseudoplusia includens larvae indicated that copy number of MdBV segments C, B and J varied between hosts but their relative abundance within a host was virtually identical to their abundance in calyx fluid. Among-tissue assays indicated that each viral segment was most abundant in hemocytes and least abundant in salivary glands. However, the relative abundance of each segment to one another was similar in all tissues. We also found no clear relationship between MdBV segment and transcript abundance in hemocytes and fat body.

Animals↗

Management of ectoparasites with biological control organisms.

Biological control is not a new concept, but for many reasons it is gaining interest for control of livestock ectoparasites. These reasons will be discussed, both from a political view and from environmental and economic views. The US government has vowed to reduce pesticide use by the year 2000, but other forces may drive this change even faster. Pesticide costs are high, and efficacy against some pests is questionable. Also, many producers are concerned about the environment, and are anxious to do their part to reduce chemical pollution. Specialised training is required to reduce on-the-farm difficulties involved with the use of biological-control organisms. Otherwise, how do producers or veterinarians purchase and use biocontrol organisms, and how do they critique what has been purchased? Included is a short summary of the three most common ectoparasites of livestock, and the type of biological-control strategies being developed to combat them. Much of the classical work has been done on filth fly control, most likely because of the nuisance status of flies, and because of the availability of candidate beneficial organisms, particularly parasitic wasps. And finally, two fly-control success stories will be briefly described. Tremendous strides have been made in house-fly and stable-fly control with parasitic wasps on feedlots, but more work is needed to better understand the habits of immature fly populations. A predaceous fly is being tested for pest fly control in dairies. Larvae of this fly can kill 15-20 house-fly larvae daily, and the adults do not become pestiferous on farms or around homes. Biological control will be an important part of livestock pest control in the future, but its implementation will require a corps of educated producers who are confident that biological control can work for them.

Animals↗

Expression of Cotesia kariyai polydnavirus genes in lepidopteran hemocytes and Sf9 cells.

The parasitic wasp Cotesia kariyai carries polydnavirus (CkPDV) which is an indispensable factor for the successful parasitization by the wasp. One of its surface proteins has been identified as an immunoevasive mediator from the cellular defense reactions of the host armyworm Pseudaletia separata, thereby it was named immunoevasive protein (IEP). In the present study, we demonstrated that anti-IEP antibody did not suppress the CkPDV infection of Sf9 cells but decreased its infection of P.separata hemocytes, thus indicating that IEP is not essential for CkPDV to enter into the target cells but is important for evading from the attack of the hemocytes. Three genes of CkPDV expressed in Sf9 cells were isolated and characterized. Two of them (CkV0.8, CkV0.9) are novel genes but another one (CkV2.0) is the same gene with the one we previously identified in the parasitized armyworm larvae. Although these genes reside in different DNA segments of CkPDV genome, all of them are expressed in the hemocytes of the parasitized armyworm larvae. These gene transcripts are first detected at 2 h after parasitization, and the expressions of CkV0.8 and CkV0.9 were gradually decreased after reaching the maximum level at 4 h after parasitization. However, the expression of CkV2.0 continues to be increased at least for 10 h after parasitization.

Amino Acid Sequence↗

Influence of antibiotics on the offspring production of the Wolbachia-infected parthenogenetic parasitoid Encarsia formosa.

Three different concentrations of the antibiotic tetracycline in honey were tested for their influence on the offspring production and longevity of the parasitoid wasp Encarsia formosa. Several earlier publications did not provide a conclusive answer on the effect that the Wolbachia have on these wasps. The results of our experiments show that at high tetracycline hydrochloride concentrations in honey (50mg/ml) the antibiotic is toxic to the females, all females died within three days after the antibiotic treatment. The concentration 5mg/ml was less toxic although the treated females also lived shorter and produced less offspring than the control females. At the lowest tested concentration of 1mg/ml there was no significant difference either in offspring production or in longevity between the control and the treated females. The antibiotic treatment at both 5 and 1mg/ml resulted in exclusively male progeny after the first two days of oviposition. These results are consistent with the theory that in species in which all individuals are infected the Wolbachia should not impose a large fitness cost.

Animals↗

Fig volatile compounds--a first comparative study.

We analysed the compounds of volatile blends released by receptive figs of twenty Ficus species to attract their specific pollinating wasps. In all, 99 different compounds were identified. The compounds are mainly terpenoids, aliphatic compounds and products from the shikimic acid pathway. In each species blend, there are few major compounds, which are generally common among floral fragrances. Most species blends also include rare compounds, but generally their proportion in the blend is low. A possible basis for species-specificity of Ficus-wasp interactions is discussed in relation to the patterns of volatiles found in this interspecies comparison.

Animals↗

The Hymenoptera venom study. III: Safety of venom immunotherapy.

One thousand four hundred ten (44%) of the 3236 subjects in the Hymenoptera venom study accepted venom immunotherapy (VIT). Time to maintenance averaged 95 days, and the largest number achieved maintenance (147 subjects, 10.4%) at day 56. Ninety-two percent of the treated subjects achieved maintenance, and 84% continued therapy, most subjects (91%) until the study was terminated. One hundred seventy-one subjects (12%) experienced 327 treatment systemic reactions (Srs). The incidence of pruritus and angioedema/urticaria was similar with mild, moderate, or severe SRs. The SR severity did not correlate with the severity of the most recent sting before entry into the Hymenoptera-venom study, the most severe historical sting SR, the most severe SR during venom skin tests, the total dose of venom, the degree of skin test reactivity, or the lowest concentration yielding a positive skin test. Most SRs occurred between 1 and 50 micrograms and at maintenance; honeybee or wasp venoms were most likely to produce SR. This study, the largest of its kind with the use of standardized extracts, demonstrates (1) that there was good compliance, (2) that various historical and diagnostic criteria did not predict SRs to VIT, (3) that SRs to VIT were most likely to occur between 1 and 50 micrograms and at maintenance, (4) that honeybee or wasp venoms were most likely to produce an SR, and (5) that VIT is relatively safe.

Animals↗

Evidence for integration of Glyptapanteles indiensis polydnavirus DNA into the chromosome of Lymantria dispar in vitro.

Polydnaviruses replicate within calyx cells of the female ovaries of certain species of parasitic wasps and are required for the successful parasitization of lepidopteran hosts. These viruses, which have unusual double-stranded circular DNA segmented genomes, are integrated as proviruses into the genomes of their associated wasp hosts and are believed to be transmitted vertically through germline tissue. Here, by combined Southern hybridization, polymerase chain reaction (PCR) assays and viral sequence analyses we provide evidence that DNA originating from two distinct double-stranded circular segments of the polydnavirus genome from the braconid Glyptapanteles indiensis (GiPDV) integrates in vitro into the genome of cells derived from the natural host, Lymantria dispar. The G. indiensis polydnavirus DNA, as a result of its unique ability to be integrated in part into the chromosome of cells derived from its lepidopteran host, has potential to be developed as an in vitro cell transformation system.

Animals↗

Expression and characterization of a novel teratocyte protein of the braconid, Microplitis croceipes (cresson).

Microplitis croceipes wasps overcome host immunity by inducing changes in host physiology using factors derived from the embryo and/or larva. Teratocytes of some parasitic wasps circulate in the host hemolymph after egg hatch and synthesize proteins (TSPs), some of which are secreted to alter host physiology in support of endoparasitoid development. TSPs appear to alter host physiology, at least in part, by inhibiting synthesis of certain proteins. M. croceipes teratocytes synthesize a 13.9 kDa protein (TSP14), which inhibits synthesis of host proteins that are linked to larval growth and development. A cDNA encoding TSP14 was generated by RT-PCR from teratocyte RNA, and cloned into yeast expression vectors to produce sufficient recombinant protein for functional analyses. RecTSP14 was produced using the yeast expression system at a concentration of more than 300 micrograms/L. The recTSP14 inhibited in vitro translation of larval Heliothis virescens RNA, with the activity sensitive to boiling, protein concentration, incubation time, and storage temperatures. Although recTSP14 inhibited translation of some cellular RNAs in vitro, the in vivo incorporation of [35S]-methionine into proteins of selected insect and mammalian cell lines was not inhibited. These findings suggest that recTSP14 entry is cell type-specific and required to inhibit synthesis of target protein(s).

Animals↗

Polydnavirus infection inhibits translation of specific growth-associated host proteins.

The wasp Campoletis sonorensis injects a polydnavirus (CsPDV) along with its egg during parasitization of Heliothis virescens larvae. CsPDV protects the wasp egg and larvae by selectively disabling the host's cellular immune response, and by altering host physiology, growth, and development. Among the changes in host physiology brought about by CsPDV infection is a rapid, and specific decline in the translation of fat body mRNAs encoding selected major plasma proteins. Translational inhibition of the synthesis of all storage protein monomers, p82 (Riboflavin binding hexamer), and p74/p76 (arylphorin), occurs upon infection with CsPDV. Moreover, the prewandering peak of the plasma enzyme juvenile hormone esterase (JHE) was blocked by CsPDV injection. Northern blotting of fat body mRNA demonstrated that transcript levels of storage proteins were not affected by infection. Plasma titers of the iron binding proteins transferrin (p72) and ferritin (p24/26), and of the plasma juvenile hormone binding protein (p25) were not changed by CsPDV infection. That storage protein and JHE synthesis are translationally suppressed, while the synthesis of other plasma proteins continues apace, suggests that CsPDV infection may lead to translational discrimination among available mRNAs in CsPDV infected fat bodies. The effect of this translational discrimination is to shunt host resources away from larval growth and adult development, which presumably makes them available to the developing endoparasitoid.

Animals↗

A comparative study of the proventricular structure in corbiculate apinae (Hymenoptera, Apidae).

The present study compares the proventricular structure, analyzed under scanning electronic microscope (SEM), among tribes of corbiculate Apinae. Fifty-one species of stingless bees (Meliponini), one species of honeybee (Apini), three species of bumblebees (Bombini) and seven species of orchid bees (Euglossini), were analyzed as in-group, and one species of sphecid wasp (Sphecidae) and two species of Halictidae bees, as out-groups. The proventricular bulb presents a basic morphology pattern similar to that of other Hymenoptera such as ants and wasps, being a symplesiomorphy for bees. The shape of proventricular folds constitutes a synapomorphy for Meliponini and an autapomorphy for Apini. The shape of hair-like projections of the cuticle that lines the proventriculus is a synapomorphy for Meliponini and Apini. These proventricular data corroborate the monophyly of the tribe Meliponini and the hypothesis that recognizes only one tribe for stingless bees. In addition, Meliponini+Apini constitutes a monophyletic group and Bombini+Euglossini another monophyletic group. The results confirm that internal morphology is a character that can be used in studies of the phylogeny in insects and the use of SEM as a powerful tool in these analyses.

Animals↗