Solana Abraham

DEMOGRAPHIC AND QUALITY CONTROL PARAMETERS OF ANASTREPHA FRATERCULUS (DIPTERA: TEPHRITIDAE) MAINTAINED UNDER ARTIFICIAL REARING

Abstract The integration of the sterile insect technique (SIT) in the management of the South American fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) is a promising alternative to chemically-based control in those areas where it is sympatric with Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) or other tephritid species for which the SIT is being used. Implementation of the SIT requires the development of a cost effective mass-rearing protocol. In this work, we present demographic and quality control parameters for the A. fraterculus strain reared at the Estación Experimental Agroindustrial Obispo Colombres, Tucumán, Argentina. Considering the rearing cage as the reproduction unit, we observed that fecundity is optimal during the first 3 weeks after the onset of oviposition. Fertility was constant during this period. During 2003 and 2004, some improvements were made to the existing rearing protocol, which resulted in increased larval viability, pupal weight, and adult emergence. Current weekly egg production is 1 million per week. These eggs are used to maintain the colony and to assess quality parameters. Finally, research needs leading to improved yields and fly quality are discussed.

Evolution of pre-zygotic and post-zygotic barriers to gene flow among three cryptic species within the Anastrepha fraterculus complex

Tropical tephritids are ideally suited for studies on population divergence and speciation because they include species groups undergoing rapid radiation, in which morphologically cryptic species and sister species are abundant. The fraterculus species group in the Neotropical genus Anastrepha is a case in point, as it is composed of a complex of up to seven A. fraterculus morphotypes proposed to be cryptic species. Here, we document pre‐ and post‐zygotic barriers to gene flow among adults of the Mexican A. fraterculus morphotype and three populations (Argentina, Brazil, and Peru) belonging to two separate morphotypes (Brazilian 1 and Peruvian). We unveiled three forms of pre‐zygotic reproductive isolation resulting in strong assortative mating. In field cages, free‐ranging male and female A. fraterculus displayed a strong tendency to form couples with members of the opposite sex belonging to their own morphotype, suggesting that male pheromone emission, courtship displays, or both intervene in shaping female choice before actual contact and coupling. In addition, males and females of the Peruvian morphotype became receptive and mated significantly later than adults of the Mexican and Brazilian 1 morphotypes. After contact, Mexican females exhibited greater mating discrimination than males when facing adults of the opposite sex belonging to either the Peruvian or the Brazilian 1 morphotype as evidenced by vigorous resistance to penetration once they had been forcefully mounted by heterotypic males. Forced copulations resulted in production of F1 hybrids that were either less viable (and partially fertile) than parental crosses or even sterile. Our results suggest that the Mexican morphotype is a distinct biological entity and that pre‐zygotic reproductive isolation through divergence in courtship or male‐produced pheromone and other mechanisms appear to evolve faster than post‐zygotic isolation in the fraterculus species group.

Ongoing speciation within the Anastrepha fraterculus cryptic species complex : the case of the Andean morphotype

The Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) cryptic species complex is currently composed of seven taxonomically recognized morphotypes. Both, pre‐ and post‐zygotic isolation has been documented among four of these morphotypes, revealing that in fact they appear to be distinct biological entities. In order to progress in the full delimitation of species within the complex, we examined reproductive isolation between a Colombian population of the Andean morphotype and populations belonging to four other morphotypes spanning from Mexico to Argentina. Flies from the Andean morphotype exhibited strong pre‐zygotic mating isolation through temporal partitioning of mating activity. Post‐zygotic isolation was observed for crosses of males of all morphotypes and Andean morphotype females, yet most of the F1 hybrid ♂ × F1 hybrid ♀ self‐crosses showed normal levels of fertility, a finding suggesting a nuclear–cytoplasmic interaction according to previous studies. Overall, the Andean morphotype within the complex also appears to be a distinct biological entity. We discuss the implications of these findings for the understanding of speciation mechanisms in the Neotropical genus Anastrepha.

Effects of male nutrition on sperm storage and remating behavior in wild and laboratory Anastrepha fraterculus (Diptera: Tephritidae) females

Male physiological condition can affect his ability to modulate female sexual receptivity. Thus, studying this aspect can have biological and practical implications. Here, we examine how male nutritional status affected the amount of sperm stored, remating rate and refractory period of the tephritid fruit fly Anastrepha fraterculus (Wiedemann) females. Both wild and laboratory flies were evaluated. We also examine female sperm storage patterns. Experiments were carried out by manipulating male adult diet and exposing these males to virgin females. Females mated with differently treated males were either dissected to count the amount of sperm stored or exposed to virgin males to determine remating rate and the length of the refractory period. We found that male nutritional status affected the amount of the sperm stored and the renewal of sexual receptivity in wild flies. For laboratory flies, male nutritional status affected the length of the refractory period but not the amount of sperm stored by females. In addition, we report that the ventral receptacle is not an important organ of sperm storage in this species. We conclude that male nutritional condition influences the ability to modulate female sexual receptivity, possibly through a combination of the quantity and quality of the ejaculate. From an applied perspective, providing males with an enriched diet will likely result in increased efficacy of the sterile insect technique.

Multiple mating in Anastrepha fraterculus females and its relationship with fecundity and fertility

The occurrence of female remating has been widely reported in insects and the frequency at which it occurs and the factors driving females’ remating behavior have been shown to be both species specific and variable within species. Herein, we studied the remating behavior of females from a well established laboratory colony and a wild population of the South American fruit fly, Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae), under laboratory conditions. Latency to first mating (number of days from the onset of the experiment until the first copula) was shorter for remating females than for females that did not remate. Two‐day fecundity was higher for females that did remate than for monogamous females. Egg hatch was sustained after remating and was not affected by the number of times the female mated. However, when females willing to remate were prevented from doing so, percent egg hatch showed a significant drop. These results and the fact that remating occurred more often in more fecund females than in less fecund ones suggest that remating may be a response to sperm depletion. Remating frequency was similar in laboratory and wild flies, but 2‐day fecundity was higher for laboratory than for wild females of similar mating status. Also, the length of the refractory period (time between first and second copulation) was longer for wild than for laboratory females. Differences between strains could be the result of artificial selection. Results are discussed from a theoretical and applied perspective in the context of direct benefits to females.

Fruit compounds affect male sexual success in the South American fruit fly, Anastrepha fraterculus (Diptera: Tephritidae)

Although exposure to plants has been shown to influence sexual behaviour in a number of phytophagous insect species, a relatively small number of fruit flies have been investigated in that respect. Here, we evaluated the effect of exposure to the pulp of guava and mango and to essential oils emanating from glands in the flavedo area of lemons on the mating success of Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) males. We also evaluated different durations of exposure and the need for physical contact with the fruit. Results showed that exposure to guava increased the mating success of both wild and laboratory males relative to non‐exposed males. In addition, exposed wild flies copulated earlier than non‐exposed males. Physical contact with the fruit or ingestion of compounds was not a prerequisite for this phenomenon to occur, since just the exposure to volatiles resulted in a significant enhancement in mating success. Exposure to mango did not affect male sexual performance. In contrast, exposure to lemon for 1 day decreased mating success of males relative to unexposed males, whereas exposure for longer periods seemed to eliminate this effect resulting in comparable mating success rates between treated and untreated males. These results provide a better understanding of the sexual behaviour of this species and may also find application for its control.

Female receptivity in Anastrepha ludens (Diptera: Tephritidae) is not modulated by male accessory gland products

In numerous insects, accessory gland products (AGPs) transferred from males to females during mating are responsible for female sexual inhibition, but these products can be affected by male condition. Here, we investigated the effect of AGPs on female receptivity of the Mexican fruit fly Anastrepha ludens (Loew), and the effect of male and female strain, male irradiation, AGP dose and sexual activity period on the effectiveness of these AGPs in inhibiting female remating. Injections of aqueous extracts of male accessory glands into the abdomen of females did not reduce their receptivity either at 0.2 or 0.8 male equivalent. Females injected with AGPs behaved like virgin females and not as mated females. Neither male origin, female origin (wild versus mass-reared), nor male irradiation (sterile versus fertile males) had an effect in inhibiting female remating. Also, injections of glands obtained during the sexual calling period of males, or obtained during the morning when males are not sexually active had no effect on female remating behavior. Mated mass-reared females were more likely to remate than wild females. We conclude that inhibition of female sexual receptivity of A. ludens is mediated by factors other than AGPs, such as the number of sperm stored by females, the stimulus of copulation per se or more probably, mediated by a combination of factors. More research is needed to elucidate the role of AGPs in this species.

Development of a larval diet for the South American fruit fly Anastrepha fraterculus (Diptera: Tephritidae)

Mass-rearing protocols must be developed, in particular, a cost-effective larval diet, to implement the sterile insect technique against Anastrepha fraterculus (Wiedemann). The key elements of this diet are the optimal nutrients and their concentrations, diet supports or bulking agents, and the pH of the diet. To improve the currently used formulation and develop a more cost-effective diet, in the present study, we evaluated various supports, sugar contents, nutrient proportions and pH levels. Egg-to-pupa recovery was found to increase when agar was replaced with sponge cloth in the diet. Although low, the recovery values obtained when using the sugarcane bagasse-based diet were similar to those obtained when using the agar-based diet, but the percentage of adult emergence was lower. Larval viability was found to increase when the amount of sugar in the diet was doubled. Yet, it is still necessary to evaluate this diet on a larger scale and determine the feasibility of reusing the sponge cloth to reduce the costs of larval diets.

Female Age Determines Remating Behavior in Wild Mexican Fruit Flies

Male and female age are important factors that can influence mating and remating behavior. Females can discriminate against or prefer older males, but there have been relatively fewer studies on how female and male age influence female remating. Here we showed in wild flies of the Mexican fruit fly Anastrepha ludens (Loew), that when females were given a choice between males of different ages, younger females preferred to mate with younger males over older males, while older females were less selective. Also, when given a choice between males of different ages, older females had longer copulation durations than younger females. On the other hand, older males and females had lower mating success, compared with young and middle-aged flies under no choice conditions. However, middle-aged females mated faster compared to young females and young males mated faster compared to middle-aged males. Male age did not influence female remating, while female age strongly determined female remating, with no females remating when they were old. It is unclear if female receptivity mechanisms are switched off at older ages, or if females are reluctant to remate due to possible costs of mating. We discuss our results in terms of how male and female age can influence mating decisions.

Male age and strain affect ejaculate quality in the Mexican fruit fly: Age and strain affect Mexican fly ejaculates

Aging in all organisms is inevitable. Male age can have profound effects on mating success and female reproduction, yet relatively little is known on the effects of male age on different components of the ejaculate. Furthermore, in mass‐reared insects used for the Sterile Insect Technique, there are often behavioral differences between mass‐reared and wild males, while differences in the ejaculate have been less studied. The ejaculate in insects is composed mainly of sperm and accessory gland proteins. Here, we studied how male age and strain affected (i) protein quantity of testes and accessory glands, (ii) the biological activity of accessory gland products injected into females, (iii) sperm viability, and (iv) sperm quantity stored by females in wild and mass‐reared Anastrepha ludens (Diptera: Tephritidae). We found lower protein content in testes of old wild males and lower sperm viability in females mated with old wild males. Females stored more sperm when mated to young wild males than with young mass‐reared males. Accessory gland injections of old or young males did not inhibit female remating. Knowledge of how male age affects different ejaculate components will aid our understanding on investment of the ejaculate and possible postcopulatory consequences on female behavior.