Norikuni Kumano

Biological Role of Nardonella Endosymbiont in Its Weevil Host

Weevils constitute the most species-rich animal group with over 60,000 described species, many of which possess specialized symbiotic organs and harbor bacterial endosymbionts. Among the diverse microbial associates of weevils, Nardonella spp. represent the most ancient and widespread endosymbiont lineage, having co-speciated with the host weevils for over 125 million years. Thus far, however, no empirical work on the role of Nardonella for weevil biology has been reported. Here we investigated the biological role of the Nardonella endosymbiont for the West Indian sweet potato weevil, Euscepes postfasciatus. This insect is an experimentally tractable pest insect that can easily be reared on a natural diet of sweet potato root as well as on an agar-based artificial diet. By larval feeding on an antibiotic-containing artificial diet, Nardonella infection was effectively eliminated from the treated insects. The antibiotic-treated insects exhibited significantly lighter body weight and lower growth rate than the control insects. Then, the antibiotic-treated insects and the control insects were respectively allowed to mate and oviposit on fresh sweet potatoes without the antibiotic. The offspring of the antibiotic-treated insects, which were all Nardonella-negative, exhibited significantly lighter body weight, smaller body size, lower growth rate and paler body color in comparison with the offspring of the control insects, which were all Nardonella-positive. In conclusion, the Nardonella endosymbiont is involved in normal growth and development of the host weevil. The biological role of the endosymbiont probably underlies the long-lasting host-symbiont co-speciation in the evolutionary course of weevils.

Effect of irradiation on mating ability in the male sweetpotato weevil (Coleoptera: Curculionidae).

Abstract The sterile insect technique (SIT) is widely used for suppressing or eradicating target pest insect populations. The effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females. Irradiation is the effective manner to sterilize mass-reared insects. The negative impacts of this procedure are not limited to damage on reproductive cells. Gamma-radiation damages the epithelial tissue of midgut, which affects the alimentation in insects. Irradiated males alter their mating behavior over time because of the depression of metabolic activity by sterilization. In this study, we evaluated the male mating performance and sexually compatibility of irradiated male Cylas formicarius elegantulus (Summers) (Coleoptera: Curculionidae) with a 200-Gy dose, as currently used in the SIT program in Okinawa Prefecture, throughout 16 d after irradiation in the laboratory. The mating ability of irradiated males did not differ from that of control males for about a week. However, the mating ability of irradiated male drastically decreased thereafter. We consider that irradiated male C. formicarius elegantulus with a 200-Gy dose had no major effect on male mating behavior approximately for a week after irradiation.

Copulation reduces the duration of death-feigning behaviour in the sweetpotato weevil, Cylas formicarius

Although there have been numerous studies on the effects of mating history on mating behaviour, few studies have reported the relationship between mating history and other contextual behaviours such as foraging and predator avoidance. We examined the effect of mating history on death-feigning behaviour (an antipredator behaviour) in the sweetpotato weevil. Because mating behaviour can be divided into phases, we examined the effects of encounters with the opposite sex, copulation and insemination success on death-feigning behaviour. For females after copulation and males after multiple copulations the duration of death-feigning behaviour was reduced, whereas encounters with the opposite sex had no effect. Insemination success did not affect the duration of death feigning in males, but inseminated females reduced the duration of death feigning. We discuss the implications of these results for the effect of mating history on this antipredator behaviour.

The neogregarine protozoan Farinocystis sp. reduces longevity and fecundity in the West Indian sweet potato weevil, Euscepes postfasciatus (Fairmaire)

The number of West Indian sweet potato weevils, Euscepes postfasciatus, being mass-reared in a facility for use in sterile insect technique (SIT) eradication programs has undergone a drastic reduction. A neogregarine protozoan pathogen Farinocystis sp. (an undescribed species) was detected in vivo in the mass-reared E. postfasciatus. We investigated the effects of this disease on the longevity and fecundity of host weevils and the incubation time of the disease in the host body under mass-rearing conditions. Our results demonstrated that infection by this Farinocystis sp. decreased both longevity and fecundity in E. postfasciatus. In particular, the pathogen severely limited the production of progeny by infected females compared to healthy females. Therefore, we consider this protozoan infection to be the major cause of the decreased E. postfasciatus production in the mass-rearing facility.

Effect of body size and sex ratio on male alternative mating tactics of the West Indian sweetpotato weevil, Euscepes postfasciatus.

Male body size is considered to be one of the major determinants of mating success among many insect species. Because the effectiveness of the sterile insect technique (SIT) depends on the ability of released sterile males to mate with and inseminate wild females, it is indispensable to understand the effect of male body size on the mating behavior of both sexes for the progress of the SIT program. We investigated how male body size and the presence of other rival males affect the guarding and copulatory durations of the West Indian sweetpotato weevil, Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae). In this species, males guard females before and after copulation. By observing the mating behavior under two sex‐ratio conditions (male‐to‐female ratios of 1:1 and 2:1), we found that small males hastened to court females when rival males were present, but the females rejected these small males as mates. Therefore, we consider that female weevils adopt a counter‐adaptation for mate preference in response to this male mating strategy. Body size did not affect the durations of copulation and post‐copulatory guarding. Although we found a conditional mating strategy for body size in E. postfasciatus, it is unlikely to have a large influence on the weevil‐eradication program using SIT.

Inbreeding avoidance or tolerance? Comparison of mating behavior between mass-reared and wild strains of the sweet potato weevil

Inadvertent selection is an important genetic process that frequently occurs during laboratory culture. The mass-reared strain of the sweet potato weevil Cylas formicarius exhibits stronger inbreeding depression than the wild strain does. When inbreeding depression occurs in a population, mating with a close relative is often considered maladaptive; however, in some contexts, the inclusive fitness benefits of inbreeding may outweigh the costs, favoring individuals that tolerate a low level of inbreeding depression. Theory predicts that mass-reared strain weevils will avoid inbreeding while wild strain weevils will tolerate inbreeding. To examine this prediction, we compared the effect of relatedness on the mating and insemination successes in mass-reared and wild strains of C. formicarius. While close relative pairs of the wild strain copulated less frequently than non-kin pairs, almost all mass-reared strain pairs copulated irrespective of relatedness. The results showed that the strain with weak inbreeding depression (wild strain) avoided inbreeding, whereas the strain with strong inbreeding depression (mass-reared strain) tolerated inbreeding. The contradiction between the theoretical prediction and our results is discussed from the perspective of laboratory adaptation, mating systems, and life history of C. formicarius.

Effect of Mass Rearing on Life History Traits and Inbreeding Depression in the Sweetpotato Weevil (Coleoptera: Brentidae)

ABSTRACT Inadvertent selection is an important genetic process that frequently occurs during laboratory culture and maintenance of biological control agents and other beneficial organisms used in procedures such as the sterile insect technique (SIT). We investigated effects of mass rearing and inbreeding depression on life history traits (number of progeny emerging from host plants, body weight, developmental period, and starvation tolerance) in the sweetpotato weevil, Cylas formicarius elegantulus (Summers) (Coleoptera: Brentidae). The effect of inbreeding was measured by comparing the results obtained from the full-sib crosses with those obtained from nonkin crosses in both wild and mass-reared strains. The mass-reared strain had more progeny than the wild strain. The developmental period of mass-reared strain was shorter than that of the wild strain. Other traits did not differ significantly between strains. We detected inbreeding depression effects on numbers of progeny, and the effects were more pronounced in the mass-reared strain. Hence, laboratory adaptation to mass rearing can produce changes in important biological attributes of sweetpotato weevils.

Female mating status does not affect male mating behavior in the West Indian sweetpotato weevil, Euscepes postfasciatus

Because multiple mating by females encourages sperm competition, the assessment of female mating status before insemination is important for males in order to avoid the risk of sperm competition or to intensify sperm competition. When interacting with females before sperm transfer, males can alter their mating tactics according to the risk or intensity of sperm competition. Information on how mating systems are associated with sperm competition is essential for sterile insect technique eradication programs, which depend on successful mating of released sterile males with wild females. We tested whether males of the West Indian sweetpotato weevil, Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae), adjusted their mating behavior in response to female mating experience and/or age. As virgin/young female weevils accepted males more easily than non‐virgin/old females, assessing females before insemination can be adaptive for male weevils. We found that E. postfasciatus males were unable to adjust their mating tactics in response to female mating status. Although this mating strategy would be costly for individual male weevils, the ability in males to discriminate female mating status can prevent the chance of mating with already‐mated females containing the sperm of wild male(s). Therefore, the mating tactics of male E. postfasciatus are advantageous for sterile insect technique eradication programs.

Intensive resistance by females before copulation induces insemination failure in the West Indian sweet potato weevil Euscepes postfasciatus

Persistent mating attempts by males (sexual harassment) are frequently observed among animals. For females, resisting persistent males can be costly because vigorous resistance increases both energy expenditure and the possibility of injury. Although one tactic for coping with male harassment is to cease resistance and mate with the persistent partner, the females of several species are able to prevent the fertilization of their egg(s) despite copulation. In this study, we used three different sex ratios to investigate whether a male’s mating persistence affects his mating success in the West Indian sweet potato weevil Euscepes postfasciatus, in which males mount females both before and after copulation. Consistent with our predictions, we found that female weevils resist and manipulate sperm transfer either before or during copulation according to their preferences. Female weevils were able to reject the sperm of persistent males despite having copulated with them. However, neither copulation and/or post-copulatory mounting affected insemination success. We speculate that the intensive resistance shown by females before copulation may induce mechanical sterility in E. postfasciatus.

Prolongation of the effective copulation period by fractionated-dose irradiation in the sweet potato weevil, Cylas formicarius

The sterile insect technique (SIT), based on the principles of population and behavioral ecology, is widely used to suppress or eradicate target pest insect populations. The effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females; however, the use of gamma radiation to induce sterility negatively affects both somatic cells as well as reproductive cells. Consequently, sterilization by irradiation drastically diminishes mating performance over time. It is well known that fractionated‐dose irradiation, in which a sterilizing dose is delivered via a series of smaller irradiations, reduces radiation damage. In the present study, we evaluated the effect of fractionated‐dose irradiation on fertility, longevity, and mating propensity in Cylas formicarius (Summers) (Coleoptera: Brentidae) for 16 days after irradiation. Fractionated‐dose irradiation with 200 Gy induced full sterility regardless of the number of radiation doses. Although the mating propensity of males sterilized by a single 200 Gy dose (the current standard of the Okinawa Prefecture SIT program) was equal to that of non‐irradiated weevils for the first 6 days, the mating propensity of males sterilized by a series of three doses was maintained for at least the first 12 days. These results demonstrated that fractionated‐dose irradiation can be highly advantageous in C. formicarius eradication programs.