No evidence for Wolbachia‐induced parthenogenesis in the social Hymenoptera

In some parasitoid wasps, infection by the micro‐organism Wolbachia leads to asexual reproduction. Within the Hymenoptera, the limits of distribution of parthenogenesis inducing Wolbachia have not yet been established. To address this issue, we screened all known thelytokous social hymenopteran species using a PCR assay. None was infected, and therefore we conclude that worker thelytoky evolves independently of Wolbachia in ants and bees. This supports the previously proposed hypothesis that a sex determining system based on heterozygosity may form a proximate limitation to Wolbachia‐induced parthenogenesis.     

Comparison of Hemolymph and Holotissues of Different Species of Insects as Diet Components forin VitroRearing ofTrichogramma galloiZucchi andT. pretiosumRiley

The aim of this work was to compare pupal holotissues (Diatraea saccharalis, Anticarsia gemmatalis,andBombyx mori) and larval hemolymph of several moth species (Helicoverpa zeaandBombyx mori) commonly produced in Brazilian laboratories as components of artificial diets forin vitrorearing ofTrichogramma galloiandTrichogramma pretiosum.Diets based on larval hemolymph ofH. zeaproduced the best development of both natural enemies, although a diet with pupal holotissues ofB. moriallowed full development ofT. pretiosum.Amino acid analysis showed qualitative differences among the insect derivates and eggs of the natural and alternative hosts, but with a quantitative similarity. Electrophoretic analysis resulting in a diverse protein-stained bands evidenced the qualitative differences among these components.     

Manipulation of oviposition choice of the parasitoid wasp, Encarsia pergandiella, by the endosymbiotic bacterium Cardinium

Reproductive manipulations of hosts by maternally inherited bacterial endosymbionts often result in an increase in the proportion of infected female hosts in the population. When this involves the conversion of incipient males to genetic or functional females, it presents unique difficulties for symbionts invading hosts with sex-specific reproductive behaviours, such as the autoparasitic Encarsia pergandiella. In sexual forms of this species, female eggs are laid in whitefly nymphs and male eggs are laid in conspecific or heterospecific parasitoids developing within the whitefly cuticle. Further, eggs laid in the 'wrong' host do not ordinarily complete development. This study explored the role of a bacterial symbiont, Cardinium, in manipulating oviposition behaviour in a thelytokous population of E. pergandiella. Oviposition choice was measured by the number and location of eggs deposited by both infected and uninfected adult waSPS in arenas containing equal numbers of hosts suitable for the development of male and female waSPS. Uninfected waSPS included antibiotic-treated female waSPS and (untreated) daughters of antibiotic-treated female waSPS. The choices of waSPS in the thelytokous population treatments were compared with those of a conspecific sexual population. We found that offspring of antibiotic-cured thelytokous waSPS reverted to the behaviour of unmated sexual waSPS, laying their few eggs almost exclusively in hosts appropriate for male eggs. Infected thelytokous waSPS distributed their eggs approximately evenly between host types, much like mated sexual female waSPS. The antibiotic-treated female waSPS exhibited choices intermediate to waSPS in the other two treatments. The change in the observed behaviour appears sufficient to allow invasion and persistence of Cardinium in sexual populations. Lastly, our results suggest a reduction in host discrimination as a possible mechanism by which Cardinium influences this change.     

Antibiotic curing of parthenogenesis in Eretmocerus mundus (Australian parthenogenic form)

The thelytokous parasitoid, Eretmocerus mundus (Australian parthenogenetic form) was infected with a species of Group B Wolbachia. Antibiotic treatment of the parasitoid cured parthenogenesis and lead to male production and reduced egg hatching by virgin females. For each generation infected females produced more progeny than cured females. The descendants of individuals treated with antibiotic were in most cases free of a detectable infection. Both treated females and descendant females mated and received sperm, but these were not used as mated and unmated cured females produced offspring with a similar sex ratio. Males produced as a result of the antibiotic treatment were at least partially functional as they produced sperm and copulated with and inseminated females. However, no sexual line could be established.     

A quantitative study of worker reproduction in queenright colonies of the Cape honey bee, Apis mellifera capensis

Reproduction by workers is rare in honey bee colonies that have an active queen. By not producing their own offspring and preventing other workers from producing theirs, workers are thought to increase their inclusive fitness due to their higher average relatedness towards queen-produced male offspring compared with worker-produced male offspring. But there is one exception. Workers of the Cape honey bee, Apis mellifera capensis, are able to produce diploid female offspring via thelytokous parthenogenesis and thus produce clones of themselves. As a result, worker reproduction and tolerance towards worker-produced offspring is expected to be more permissive than in arrhenotokous (sub)species where worker offspring are male. Here we quantify the extent to which A. m. capensis workers contribute to reproduction in queenright colonies using microsatellite analyses of pre-emergent brood. We show that workers produced 10.5% of workers and 0.48% of drones. Most of the workers' contribution towards the production of new workers coincided with the colonies producing new queens during reproductive swarming.     

Response of the egg parasitoid Trichogramma pretiosum to the sex pheromone of its host Heliothis zea

This paper presents results of olfactometer experiments with the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) and its host the corn earworm moth, Heliothis zea (Boddie) (Lepidoptera: Noctuidae). The sex pheromone of the host significantly reduced the total number of border crossings between odour fields in the olfactometer. Also, female parasitoids made significantly more visits to the calling moth odour field than to the opposite control field in the olfactometer. Further, the wasps spent significantly more time in the olfactometer field containing the sex pheromone released by calling virgin moths, than in control fields. If non-calling virgin moths were used as odour source, the response was reversed and wasps were repelled by the odour of the moths, and the numbers of visits were evenly distributed over the four flow fields. These results are discussed in the context of foraging ecology of egg parasitoids.

---

Résumé Des expériences menées en olfactométrie avec le parasite oophage Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) et son hôte, Heliothis zea (Boddie) (Lepidoptera: Noctuidae) donnent les résultats suivants. La présence de la phéromone sexuelle de l'hôte réduit de façon significative le nombre de passages entre les champs odorisés. De mème, les parasites visitent plus fréquemment le champ qui dispense l'odeur de la femelle en appel que le champ témoin placé à l'opposé. Par ailleurs, le temps passé dans le champ qui contient la phéromone sexuelle émise par les femelles vierges en appel est significativement supérieur à la durée de visite des champs témoins. Si l'on utilise comme source d'odeur des femelles vierges qui ne sont pas en appel, la réponse est inversée, les parasites sont alors repoussés par l'odeur de ces papillons et le nombre de visites est distribué de façon aléatoire entre les quatre champs. Ces résultats sont discutés dans le contexte de l'écologie du comportement de recherche chez les parasites oophages.

     

卷蛾赤眼蜂体内共生菌Wolbachia对寄主产雌孤雌生殖行为的影响

在30℃的条件下,连续饲养卷蛾赤眼蜂Trichogramma cacoeciaeMarchall10代,观察每代蜂的生物学特性,并进行Wolbachia的wsp基因的分子检测,以研究卷蛾赤眼蜂产雌孤雌生殖行为与其体内共生菌Wolbachia的关系。结果表明,高温下卷蛾赤眼蜂体内的Wolbachia被抑制,4代以后wsp基因的PCR检测结果变为阴性;同时赤眼蜂的产雌孤雌生殖行为的改变程度也随着世代的增加呈递增趋势。这显示卷蛾赤眼蜂的产雌孤雌生殖行为与其体内的共生菌Wolbachia具有一定程度的相关性。     

Production of females by parthenogenesis in the ant,Cerapachys biroi

Summary Groups of virginCerapachys biroi workers produced females (workers) in the laboratories. This is the first report of thelytoky in Cerapachyinae.     

In vitro culture of Trichogramma pretiosum on an artificial medium

The composition of an artificial medium and environmental conditions are described for the in vitro rearing of the egg parasite Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). The medium was composed of defined amounts of protein, carbohydrates, lipid, salts, and vitamins, but also contained up to 40% insect hemolymph. The hemolymph was necessary to induce pupation. T. pretiosum eggs were obtained by dissection of Heliothis virescens (F.) (Lepidoptera: Noctuidae) eggs. In vitro reared T. pretiosum were similar in size to H. virescens reared T. pretiosum, and females were fecund.

---

Résumé Les oeufs de Trichogramma pretiosum ont été obtenus par dissection d'oeufs d'Heliothis virescens. T. pretiosum Riley (Hymenoptère, Trichogrammatidae) a été élevé avec succès sur un substrat synthétique. Outre des quantités définies de protéines, glucides, lipides, éléments minéraux et vitamines, la ration contenait aussi jusqu'à 40% d'hémolymphe de Manduca sexta. L'hémolymphe était nécessaire pour induire la nymphose. En plus de la nourriture, les conditions d'environnement sont apparues extrêmement importantes pour élever T. pretiosum dans des conditions satisfaisantes. Le contrôle de l'humidité relative, en particulier, était le facteur le plus important. Les adultes produits au cours de cette étude étaient d'apparence normale; ils se sont accouplés sans problèmes, les femelles étaient fécondes et leur taille ne différait pas de celle d'individus élevés sur H. virescens.

     

Refinement of a cell line based artificial diet for rearing the parasitoid wasp, Trichogramma pretiosum

An artificial diet incorporating insect cells originally developed for Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae) was successfully used to rear Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). To refine the diet, individual components were removed. Chicken egg yolk and the insect cells were identified as the most important components for T. pretiosum development. Their removal resulted in few pupae and no adults. Removal of Grace’s insect medium, a common component of artificial diets, was found to markedly improve the development of T. pretiosum, producing 60% larva to pupa transition and 19% pupa to adult transition. There was no significant difference in T. pretiosum development on diets in which milk powder, malt powder or infant formula were interchanged, despite differences in nutrient composition. The use of yeast extract resulted in significantly higher survival to the adult stage when compared with yeast hydrolysate enzymatic and a combination of yeast extract and yeast hydrolysate enzymatic. Comparison of four antimicrobial agents showed the antibacterial agent Gentamycin and the antifungal agent Nystatin had the least detrimental effect on T. pretiosum development. The use of insect cell line diets has the potential to simplify artificial diet production and significantly reduce T. pretiosum production costs in Australia compared to diets using insect hemolymph or the use of natural or factitious hosts.     

Effect of photoperiod and thermoperiod on the eclosion rhythm ofTrichogramma evanescens

The dynamics of adult eclosion inTrichogramma evanescens Westw. were studied under (1) constant light and temperature of 20 °C, (2) photoperiods L12:D12 and L16:D8 at 20 °C, (3) thermoperiod 4 h 27 °/20 h 20 °C at constant light, (4) joint action of photo- and thermoperiod. The emergence was arhythmic in constant light combined with constant temperature, while a sharp monophasic rhythm was observed with the isolated action of photo- or thermoperiod. The ‘light-on’ and ‘temperature step-up’ signals were shown to act in one direction. When both signals were combined, they manifested themselveses competing entraining stimuli which, in turn, revealed an apparent individual variation in relative reactivity to the light and temperature signals. Some perspectives which follow from these observations are discussed.     

Influence of larval food consumption on the fecundity of the lacewingChrysoperla carnea

The effects of food consumption on larval growth and development and adult fecundity of the common green lacewing,Chrysoperla carnea (Stephens), were studied on two populations of larvae derived from either a laboratory colony or from field-collected adults. The number of eggs of the Mediterranean flour moth,Anagasta kuehniella (Zeller), provided to individual lacewing larvae was varied to produce three food-supply treatments: low, intermediate, and high. Food-supply was found to influence larval growth and development and adult fecundity. Lacewing larvae provided with an overabundance of moth eggs developed faster than larvae provided with fewer moth eggs than they could have consumed. Adult females that developed from the high feeding treatment had a substantially shorter preoviposition period, a later decline in egg deposition, and a significantly higher fecundity than adults arising from the other feeding classes. Unrestricted feeding by adult lacewings on an artificial diet did not compensate for prior low feeding regimes. The overall performance of the lacewings derived from the laboratory colony was substantially poorer than that of the lacewings derived from field-collected adults. This effect was accentuated when the larvae were given a low food-supply. This overall decrease in vigor is attributed to inbreeding of the laboratory culture over a one year period. These finds are relevant to mass rearing programs for this biological control agent.     

Gene flow between sexual and asexual strains of parasitic wasps: a possible case of sympatric speciation caused by a parthenogenesis-inducing bacterium

Sympatric speciation is strictly defined as the emergence of two species from a population in which mating has been random with respect to the place of birth of the mating partners. Mathematical models have shown that sympatric speciation is possible, but very few examples have been documented in nature. In this article, we demonstrate that arrhenotokous and thelytokous strains of a parasitic wasp, Neochrysocharis formosa, speciated sympatrically through infection by a symbiotic bacterium Rickettsia for the following reasons: First, Rickettsia infection was detected in all of the thelytokous strains collected throughout Japan. Second, the arrhenotokous and thelytokous strains have been collected sympatrically. Third, crossing experiments between the two strains did not result in fertilized offspring. In addition, the two strains were genetically isolated at the nuclear and mitochondrial genes. Fourth, the two strains showed a sister relationship in nuclear 28S rRNA gene. Finally, thelytokous females treated with antibiotics produced Rickettsia-free male offspring of the same reproductive form as arrhenotokous females indicating that the thelytokous strain could have speciated sympatrically from an individual of the arrhenotokous strain.     

Inter- and Intraspecific Effects of Volatile and Nonvolatile Sex Pheromones on Males, Mating Behavior, and Hybridization in Eretmocerus mundus and E. eremicus (Hymenoptera: Aphelinidae)

Eretmocerus species (Hym. Aphelinidae) are solitary parasitoids of Bemisia tabaci (Gennadius). Mate finding and mating behavior of two species, E. mundus and E. eremicus, were studied under laboratory conditions. We used three populations of Eretmocerus: typical arrhenotokous populations of E. eremicus (from USA) and E. mundus (from Spain), and an atypical thelytokous population of E. mundus (from Australia). We studied the intra- and interspecific responses of males to volatile and nonvolatile components of the female sex pheromones, mating behavior, and hybridization between populations and species. In both arrhenotokous populations, males reacted to volatile pheromones by walking toward conspecific virgin females. Males also reacted to nonvolatile pheromones by spending more time on and around patches on leaves of poinsettia plants that had been exposed to virgin females. Males of E. eremicus showed the same reaction to the nonvolatile sex pheromone of E. mundus females, but E. mundus males did not show any reaction to the nonvolatile sex pheromone of E. eremicus. There was no response of males of both species to thelytokous females of E. mundus. In both species three phases were distinguished in the mating behavior: premating, mating, and postmating. The duration of the phases differed between the three populations. Successful copulation between the two Eretmocerus species did not occur. In contrast, we recorded some successful copulations between Australian males and Spanish females of E. mundus, but they did not produce any hybrid females.     

The effect of host size on quality attributes of the egg parasitoid,Trichogramma pretiosum

In a study of the quality ofTrichogramma pretiosum Riley (Hymenoptera: Trichogrammidae), we compared female wasps emerging from natural hosts, parasitized in the laboratory or the field with those emerging from factitious hosts used for commercial mass production. Females from the natural hosts were larger, more fecund, and longer lived than those from the factitious hosts. Compared to small females, large female wasps are substantially more fecund when honey (carbohydrate) is available but marginally more fecund when honey is unavailable. The size of a femaleT. pretiosum depends on two factors: the size of the host egg from which it emerges even when the wasp was gregarious, and the number of conspecifics that emerge with it. The similarities in the size distribution of female wasps emerging from natural hosts, in conjunction with the mechanism by whichTrichogramma measure host size and allocate eggs accordingly, suggests the hypothesis that size related components of fitness in femaleT. pretiosum are under strong selection in the field.     

Biological characteristics and thermal requirements of a Brazilian strain of the parasitoid Trichogramma pretiosum reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis

A new strain of the parasitoid Trichogramma pretiosum, was collected in Rio Verde County, State of Goiás, Central Brazil, and designated as T. pretiosum RV. This strain was then found to be the most effective one among several different strains of T. pretiosum tested in a parasitoid selection assay. Therefore, its biological characteristics and thermal requirements were studied, aiming at allowing its multiplication under controlled environmental conditions in the laboratory. The parasitoid was reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis at different constant temperatures within an 18–32 °C temperature range. The number of annual generations of the parasitoid was also estimated at those temperatures. Results have shown that T. pretiosum RV developmental time, from egg to adult, was influenced by all temperatures tested within the range, varying from 6.8 to 20.3 days and 6.0 to 17.0 days on eggs of P. includens and A. gemmatalis, respectively. The emergence of T. pretiosum RV from eggs of A. gemmatalis was higher than 94% at all temperatures tested. When this variable was evaluated on eggs of P. includens, however, the figures were higher than that within the 18–30 °C range (more than 98%), and were also statistically higher than the emergence observed at 32 °C (90.2%). The sex ratio of the parasitoids emerged from eggs of A. gemmatalis decreased from 0.55 to 0.29 at 18–32 °C, respectively. However, for those emerged from eggs of P. includens, the sex ratio was similar (0.73, 0.72 and 0.71) at 20, 28 and 32 °C, respectively. The lower temperature threshold (Tb) and thermal constant (K) were 10.65 °C and 151.25 degree-days when the parasitoid was reared on eggs of P. includens; and 11.64 °C and 127.60 degree-days when reared on eggs of A. gemmatalis. The number of generations per month increased from 1.45 to 4.23 and from 1.49 to 4.79 when the parasitoid was reared on eggs of P. includens and A. gemmatalis, respectively, following the increases in the temperature.     

Use of tetracycline in larval diet to study the effect of Wolbachia on host fecundity and clarify taxonomic status of Trichogramma species in cured bisexual lines

Endosymbionts of the genus Wolbachia were efficiently cured from Trichogramma species by incorporating 0.02% tetracycline into the artificial diet used to rear larvae. Use of this technique yielded stable cured lines (bisexual and arrhenotokous lines) in which no Wolbachia organisms were detected by PCR for up to 14 generations after curing. Four cured strains of Trichogramma pretiosum showed a significantly lower total fecundity compared to their Wolbachia-infected counterpart. However, the fecundity of a single cured strain of Trichogramma evanescens was similar to its Wolbachia-infected counterpart. These differences in the effect on fecundity may be due to differences between the Wolbachia strains infecting T. pretiosum or T. evanescens, providing additional evidence for the hypothesis that a specific interaction exists between some Trichogramma species and their Wolbachia symbionts. Tetracycline in larval diet was also used to generate bisexual strains of Trichogramma oleae and Trichogramma cordubensis so that these species could be crossed with the closely related species, respectively, T. pretiosum and T. evanescens, to test their compatibility. These crosses showed a lack of compatibility, validating maintenance of these as distinct species.