The female Euscepes postfasciatus refractory period is induced by the male but length is determined by the female.

Females of many animals mate multiple times during their lives (i.e., polyandry). The period between matings (mating interval) is called the refractory period (RP). In the West Indian sweet potato weevil (Euscepes postfasciatus), males use the ejaculate to induce the RP in females to prevent re-mating. By measuring the RP, a large variation of 1-49 days was observed. This variation may be due to the males (ejaculate quantity and quality) and females (ejaculate sensitivity/degradation ability and body size) and their interactions, but the exact mechanisms are currently unclear. Here, we investigated a tendency towards a particular female RP duration and the associated traits of males and females to test the following three factors responsible for variation in the length of the RP: male manipulation of ejaculate volume, individual differences in male ejaculation substances, and ejaculate sensitivity/degradation ability in females. We prepared virgin males and females to create mating pairs. The following day, another mate was introduced to the females, and the first RP was measured. The same procedure was used for measuring the second RP. The males were also provided with another female (second female), mated, and then the RP of the second female was measured. In addition, the relationship between the length of the RP and female fitness was investigated. The results showed that there was a significant positive correlation between the first and second RP in the focal females, while no significant correlation was observed between the RP of the first and second females induced by the same male. It was also found that the length of the RP did not affect female fitness. This indicated that the males did not adaptively manipulate ejaculation volume depending on the quality of the females, and variance in the length of the RP may be explained by variation in the female physiological ability against ejaculate.

First case of successful eradication of the sweet potato weevil, Cylas formicarius (Fabricius), using the sterile insect technique.

This paper presents the first case of the successful eradication of a Coleoptera pest species over a wide area using a combination of male annihilation technique (MAT) and sterile insect technique (SIT) application. The sweet potato weevil, Cylas formicarius, is one of the most destructive and widely distributed pests of sweet potato, Ipomoea batatas. A project to eradicate it was launched in 1994 on Kume Island, Okinawa Prefecture, Japan. The MAT application was first used from November 1994 to January 1999 to reduce the density of wild populations. The distribution and densities of weevils were assessed by trapping them and surveying infestation rates in wild hosts and sweet potatoes in the field. The C. formicarius populations were suppressed by approximately 90% and plant infestations were reduced from 9.5% to less than 0.1% by using the MAT. Then, hundreds of thousands to millions of sterile weevils were released each week (ca. 460 million in total from 1999 to 2012). As a result, based on an analysis of 12748 stems and 48749 tubers, no weevil infections were detected in the stems or tubers of sweet potato since 1997. Since 2009, almost no wild weevils were captured in traps, and in wild host and sweet potato surveys no weevils have been found in any of the 580 locations and 8833 samples since October 2011. As of 28 December, 2012, C. formicarius is considered to have been eradicated from Kume Island. This paper describes the process of eradicating C. formicarius using SIT application integrated with MAT application for the first time and discusses some of the main challenges associated with the weevil eradication campaignl.

Effects of Larval Diet on the Male Reproductive Traits in the West Indian Sweet Potato Weevils Euscepes postfasciatus (Coleoptera: Curculionidae).

Larval diet significantly affects adult traits, although less is known about how they affect reproductive traits. Males of West Indian sweet potato weevil Euscepes postfasciatus deliver a remating inhibitor along with sperm to their mates during mating, leading to a refractory period (the period before females mate again). Crossing experiments were conducted using lines reared on artificial diets, including sweet potato powder (AD) or sweet potato tubers (SP) during the larval stage, and the refractory period was examined. We also examined whether the larval diet qualitatively or quantitatively altered male ejaculate. The results showed that the refractory period was significantly longer in the SP treatment than in the AD treatment for males and females. There was no significant difference in ejaculate volume. However, the number of sperm in the testes-seminal vesicles complex was significantly higher in the SP treatment. Additionally, SDS-PAGE revealed that the ejaculate was qualitatively different depending on the larval diet, and one protein of approximately 15 kDa in size was expressed only in the SP treatments. Revealing how larval diet affects reproductive traits in adult males will help shed light on the diverse evolution of insect mating systems and reproductive behavior.

Sterile males and females can synergistically suppress wild pests targeted by sterile insect technique.

The sterile insect technique (SIT) involves periodically releasing artificially sterilized insects to inhibit normal mating between wild insect pests, ultimately resulting in the eradication of wild pest populations. It has often been discussed whether releasing either one sex, mainly males, of sterile insects (i.e., a unisexual release) can enhance the pest-control effect of the SIT more than releasing both sexes (i.e., a bisexual release). We constructed a mathematical model to examine the contribution of sterile males and females to the pest-control effect and the synergy between them. We consider that males seek out and court females in accord with their own female searching ability and preference, and that females subsequently choose one male from among males courting them in accordance with their own preference. Using this model, we compared the pest-control effect of bisexual and unisexual release, focusing on the difference in mating systems of the targeted insects. We showed that for swarm-type mating systems (with few courtship chances with higher encounter rates), bisexual release was the most effective, irrespective of the relative female searching ability between wild and sterile males. In this case, sterile females indirectly reduce wild females mating with either male by absorbing courtship from both wild and sterile males. By contrast, bisexual release is the most effective for scramble-type mating systems (more courtship chances with lower encounter rates) only when the female searching ability of sterile males is lower than that of wild males. In this case, sterile females absorb courtship from males with higher searching abilities. Therefore, the net impact of sterile females depends on the difference in sexual performance between wild and sterile males. Because the sexual performance of sterile insects is often degraded during the process of sterilization, we suggest that bisexual release can be a compatible measure to efficiently suppress wild pest populations.

Effects of Storage Periods of an Artificial Larval Diet on the Yield and Quality of Mass-Reared West Indian Sweet Potato Weevil (Coleoptera: Curculionidae).

Artificial diets have been employed for the mass-rearing of numerous insects because of their ease of use and standardized quality. An ability to store artificial diets under nonrefrigerated conditions over the long term could improve the efficacy of mass-rearing systems considerably. However, it remains largely unknown how long artificial diets can be stored at such temperatures without any adverse effects on the insects reared. In this study, we investigated yield, body size, and reproductive potential of West Indian sweet potato weevil, Euscepes postfasciatus (Fairmaire), which is a major sweet potato pest, under management using the sterile-insect technique in Japan and reared using artificial diets with different storage periods (14, 28, and 42 d) at nonrefrigerated temperatures (25 ± 1°C), and compared them with those of the control (0 d). Notably, E. postfasciatus yield and reproductive potential increased significantly with an increase in storage period (28 and 42 d). Conversely, male body size decreased significantly following feeding with artificial diet stored for 42 d, when compared with the control, while there were no significant differences in female body size between the control and all the treatments. We discuss the potential causes of such varying effects between yield and body size and conclude that E. postfasciatus artificial diet can be stored for at least 28 d without any adverse effects on weevil yield and weevil quality. To the best of our knowledge, this is the first report revealing the positive effects of long-term storage of the artificial diet on mass-reared insects.

A New System for Detecting Initial Colonization by Invasive Pests and Their Locations.

Quarantine pests in plants can be a serious agricultural problem; many eradication programs using area-wide control measures have been implemented worldwide to combat this threat. Surveillance measures using sex pheromone (in general, male-attractant) traps are also widely implemented for rapid control and eradication of invasive pests. If initial pest colonization can be determined based on temporal count data of trapped insects (i.e., males), and countermeasures are applied only during colonization, costs incurred by these countermeasures would be dramatically reduced, especially in areas with frequent invasions. In this study, we developed a system to detect initial pest colonization, and to narrow down colonized regions using estimated temporal count data of the sweet potato weevil, Cylas formicarius Fabricius (Coleoptera: Curculionidae), in Tsuken Island, Okinawa, Japan. We verified the system by comparing our estimates to actual colonization data obtained via regular host plant surveys. Results indicated that our system was able to successfully detect pest colonization and estimate colonized regions. In this study, we discuss the conditions (i.e., pest biology, environment, etc.) that are optimal for application of our system.

Inbreeding tolerance as a pre-adapted trait for invasion success in the invasive ant Brachyponera chinensis.

Identifying traits that facilitate species introductions and successful invasions of ecosystems represents a key issue in ecology. Following their establishment into new environments, many non-native species exhibit phenotypic plasticity with post-introduction changes in behaviour, morphology or life history traits that allow them to overcome the presumed loss of genetic diversity resulting in inbreeding and reduced adaptive potential. Here, we present a unique strategy in the invasive ant Brachyponera chinensis (Emery), in which inbreeding tolerance is a pre-adapted trait for invasion success, allowing this ant to cope with genetic depletion following a genetic bottleneck. We report for the first time that inbreeding is not a consequence of the founder effect following introduction, but it is due to mating between sister queens and their brothers that pre-exists in native populations which may have helped it circumvent the cost of invasion. We show that a genetic bottleneck does not affect the genetic diversity or the level of heterozygosity within colonies and suggest that generations of sib-mating in native populations may have reduced inbreeding depression through purifying selection of deleterious alleles. This work highlights how a unique life history may pre-adapt some species for biological invasions.

Radiocarbon analysis reveals expanded diet breadth associates with the invasion of a predatory ant.

Invasions are ecologically destructive and can threaten biodiversity. Trophic flexibility has been proposed as a mechanism facilitating invasion, with more flexible species better able to invade. The termite hunting needle ant Brachyponera chinensis was introduced from East Asia to the United States where it disrupts native ecosystems. We show that B. chinensis has expanded dietary breadth without shifting trophic position in its introduced range. Transect sampling of ants and termites revealed a negative correlation between the abundance of B. chinensis and the abundance of other ants in introduced populations, but this pattern was not as strong in the native range. Both termite and B. chinensis abundance were higher in the introduced range than in native range. Radiocarbon (14C) analysis revealed that B. chinensis has significantly younger 'diet age', the time lag between carbon fixation by photosynthesis and its use by the consumer, in the introduced range than in the native range, while stable isotope analyses showed no change. These results suggest that in the introduced range B. chinensis remains a termite predator but also feeds on other consumer invertebrates with younger diet ages such as herbivorous insects. Radiocarbon analysis allowed us to elucidate cryptic dietary change associated with invasion success.

Limited mobility of target pests crucially lowers controllability when sterile insect releases are spatiotemporally biased.

The sterile insect technique (SIT) is a genetic pest control method wherein mass-reared sterile insects are periodically released into the wild, thereby impeding the successful reproduction of fertile pests. In Okinawa Prefecture, Japan, the SIT has been implemented to eradicate the West Indian sweet potato weevil Euscepes postfasciatus (Fairmaire), which is a flightless agricultural pest of sweet potatoes. It is known that E. postfasciatus is much less mobile than other insects to which the SIT has been applied. However, previous theoretical studies have rarely examined effects of low mobility of target pests and variation in the spatiotemporal evenness of sterile insect releases. To theoretically examine the effects of spatiotemporal evenness on the regional eradication of less mobile pests, we constructed a simple two-patch population model comprised of a pest and sterile insect moving between two habitats, and numerically simulated different release strategies (varying the number of released sterile insects and release intervals). We found that spatially biased releases allowed the pest to spatially escape from the sterile insect, and thus intensively lowered its controllability. However, we showed that the temporally counterbalancing spatially biased releases by swapping the number of released insects in the two habitats at every release (called temporal balancing) could greatly mitigate this negative effect and promote the controllability. We also showed that the negative effect of spatiotemporally biased releases was a result of the limited mobility of the target insect. Although directed dispersal of the insects in response to habitats of differing quality could lower the controllability in the more productive habitat, the temporal balancing could promote and eventually maximize the controllability as released insects increased.

Queen-specific volatile in a higher termite Nasutitermes takasagoensis (Isoptera: Termitidae).

In social insect colonies, queen-produced pheromones have important functions in social regulation. These substances influence the behavior and physiology of colony members. A queen-produced volatile that inhibits differentiation of new neotenic reproductives was recently identified in the lower termite Reticulitermes speratus. However, there are no known queen-specific volatiles of this type in any other termite species. Here, we report volatile compounds emitted by live queens of the higher termite Nasutitermes takasagoensis. We used headspace gas chromatography mass spectroscopy (HS GC-MS) to analyze volatiles emitted by live primary queens, workers, soldiers, alates, and eggs collected in a Japanese subtropical forest. Among 14 detected compounds, 7 were soldier-specific, 1 was alate-specific, 1 was egg-specific, and 1 was queen-specific. The queen-specific volatile was phenylethanol, which is different than the compound identified in R. speratus. The identification of this queen-specific volatile is the first step in determining its functions in higher termite social regulation. Comparisons of queen pheromone substances regulating caste differentiation among various termite taxa will contribute to a better understanding of the evolution of social systems in termites.

Identification of a pheromone regulating caste differentiation in termites.

The hallmark of social insects is their caste system: reproduction is primarily monopolized by queens, whereas workers specialize in the other tasks required for colony growth and survival. Pheromones produced by reigning queens have long been believed to be the prime factor inhibiting the differentiation of new reproductive individuals. However, there has been very little progress in the chemical identification of such inhibitory pheromones. Here we report the identification of a volatile inhibitory pheromone produced by female neotenics (secondary queens) that acts directly on target individuals to suppress the differentiation of new female neotenics and identify n-butyl-n-butyrate and 2-methyl-1-butanol as the active components of the inhibitory pheromone. An artificial pheromone blend consisting of these two compounds had a strong inhibitory effect similar to live neotenics. Surprisingly, the same two volatiles are also emitted by eggs, playing a role both as an attractant to workers and an inhibitor of reproductive differentiation. This dual production of an inhibitory pheromone by female reproductives and eggs probably reflects the recruitment of an attractant pheromone as an inhibitory pheromone and may provide a mechanism ensuring honest signaling of reproductive status with a tight coupling between fertility and inhibitory power. Identification of a volatile pheromone regulating caste differentiation in a termite provides insights into the functioning of social insect colonies and opens important avenues for elucidating the developmental pathways leading to reproductive and nonreproductive castes.

Males of the seed bug Togo hemipterus (Heteroptera: Lygaeidae) use accessory gland substances to inhibit remating by females.

In species in which females mate repeatedly, males can adopt several strategies to reduce the risk of sperm competition with future males. The refractory period of females significantly increased as the mating duration increased in the seed bug Togo hemipterus (Heteroptera: Lygaeidae). To elucidate the mechanisms by which mated females are inhibited from remating, we investigated the effects of male-derived substances on the inhibition of mating receptivity of virgin females by injecting the substances into their abdomens. The length of time from injection to mating in virgin females was significantly longer for females injected with accessory gland B solution than for those injected with seminal vesicle, accessory gland A, or control solutions. This is the first report showing that heteropteran males inhibit female remating by using substances from an accessory gland. We discuss and consider the adoption and evolution of this strategy by T. hemipterus males by focusing on female genitalia structures, oviposition habit, and paternity and comparing these traits with those of other heteropterids.