Evaluation of a pelleted diet for larval mass-rearing of Anastrepha ludens and Anastrepha obliqua

Most artificial diets used for mass-rearing of fruit fly larvae involve the purchase and independent weighing of each ingredient of the formulation, followed by mixing and diet preparation process. A ready-to-use formulation would avoid this time-consuming task and simplify the preparation procedure, leading to a more standardized diet. In this work, we compared life-history traits and survival and developmental parameters of Anastrepha ludens Loew and Anastrepha obliqua (Macquart) (Diptera: Tephritidae) reared on two formulations of an artificial diet. Two experiments were performed; first, a conventional non-pelleted diet was compared to a newly developed pelleted diet, which was mixed with water before being given to the larvae. The second experiment consisted of a comparison between the pelleted diet which was stirred in a mixer with water and dissolved preservatives and a pelleted diet which was only soaked in water with dissolved preservatives. We found that the pelleted diet allowed the mass-rearing of A. ludens and A. obliqua. Moreover, feeding larvae with pelleted diet led to increased weight of the larva by 7.4 and 11.8%, and of the pupa by 9.5 and 16.7% for A. ludens and A. obliqua, respectively, compared to conventional diet. For process evaluation, the number of activities, handling time, reception, storage, and larval diet preparation were recorded. The pelleted diet only needed soaking in water containing preservatives, thereby simplifying the mass-rearing process and saving time, space, and labour. Pelletizing is a versatile technology that can be adjusted for better physicochemical quality and reduced microbiological risk. This diet should be of particular interest for the mass-rearing process in ‘sterile insect technique’ applications.     

Enhancing mating performance after juvenile hormone treatment in Anastrepha fraterculus: a differential response in males and females acts as a physiological sexing system

Methoprene (a mimic of juvenile hormone) treatment can reduce the time required for sexual maturation in Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) males under laboratory conditions, supporting its use as a treatment for sterile males within the context of the sterile insect technique (SIT). We evaluated sexual behaviour, mating competitiveness of methoprene-treated males, and female readiness to mate after methoprene-treatment in field cages. The study involved two strains of A. fraterculus from Argentina and Peru, which show several polymorphisms in relation to their sexual behaviour. We also analyzed whether methoprene treatment affected male and/or female behaviour in the same way in these two strains. Methoprene-treated males were equally competitive with untreated mature males, and became sexually competitive 6 days after emergence (3–4 days earlier than untreated males). In contrast, methoprene did not induce sexual maturation in females or, at least, it did not induce a higher rate of mating in 7-day-old females. These results were observed both for the Argentina and the Peru strains. Altogether, our results indicate that methoprene treatment produces sexually competitive males in field cages. In the absence of a genetic sexing system, and when sterile males and females of A. fraterculus are released simultaneously, the fact that females do not respond as do males to the methoprene treatment acts as a physiological sexing effect. Therefore, in the presence of mainly sexually immature sterile females, released sexually mature sterile males would have to disperse in search of wild fertile females, thereby greatly reducing matings among the released sterile insects and thus enhancing sterile insect technique efficiency.     

Sexual competitiveness of a transgenic sexing strain of the Mediterranean fruit fly, Ceratitis capitata

The sterile insect technique (SIT), when used for the control of the Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), generally relies on the release of sterile flies of only the male sex. Male selection is achieved through the use of a genetic sexing strain (GSS) in which females are killed by heat treatment in the generation prior to release. Transgenic sexing strains (TSS) have been developed that perform the same function of female-lethality, this time by withholding tetracycline (or related compounds) from the larval diet. The use of TSS may allow for certain problems associated with conventional GSS, such as strain instability and reduced productivity in mass-rearing, to be avoided. The performance, and principally the sexual competitiveness, of released male flies is important for the success of an SIT control programme. This study describes field cage experiments in which the competitiveness of males from a TSS (OX3376B) was compared with that of a conventional GSS (VIENNA-8) and two wild-type strains (TOLIMAN and ARG). When competing for female mates with wild-type males, OX3376B male performance was acceptable. When OX3376B males competed directly for mates with VIENNA-8 males, VIENNA-8 slightly outperformed the TSS males. Parallel tests, in which wild-type males competed with either OX3376B or VIENNA-8 males, showed that males from both sexing strains were highly competitive with wild-type males. These results suggest that OX3376B in particular, and TSS in general, show sufficiently good mating competitiveness to merit further research into their suitability for eventual use in SIT programmes.     

Seasonal variation of the capture and field distribution of sterile Anastrepha ludens in Tamaulipas,Mexico

Anastrepha ludens (Loew) (Diptera: Tephritidae) is one of the most important citrus pest in Mexico. The sterile insect technique (SIT) is used against pest populations of fruit flies for suppression, eradication, containment and prevention to reduce damages in fruit‐growing areas. In this study, we analyzed the seasonal variation of captures and field distribution of sterile A. ludens released in different seasons of the year in north‐eastern Mexico. Chilled releases were conducted by air at constant densities per ha on a citrus area for a period of 32 weeks that included the coldest and warmest seasons that is winter, spring and summer. Multilure traps baited with torula yeast pellets were used to capture sterile flies. Fly capture data were compared over the three seasons and correlated with climate. The lowest number of captures of the sterile insect occurred in the summer and the highest in winter and spring. High and low temperatures were negatively correlated with fly captures. Field distribution was also negatively correlated with high temperatures in summer, but no relationships were observed in winter and spring. No relationships were observed between rainfall with capture and field distribution of sterile flies. These results indicate that summer is a season involving agro‐ecological and environmental constraints for the capture and field distribution of sterile flies. This study may be useful for enhancing release strategies and optimizing economic resources in north‐eastern Mexico. Further research on the behaviour of sterile flies under stressful environments is suggested.     

Pre-release feeding on yeast hydrolysate enhances sexual competitiveness of sterile male Queensland fruit flies in field cages

Recent laboratory studies of mass‐reared flies in small cages have found that periods of just 24‐ or 48‐h access to yeast hydrolysate can substantially enhance mating performance of mass‐reared male Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (‘Q‐flies’). Using field cage tests that provide a better approximation of nature, we here investigated whether access to yeast hydrolysate for 48 h after adult emergence improves the ability of male and female mass‐reared, sterile Q‐flies to compete sexually with wild‐type flies that had been provided continuous access to yeast hydrolysate. Mating probability of sterile males was significantly increased by 48‐h access to yeast hydrolysate; sterile males provided 48‐h access to yeast hydrolysate had mating probability similar to that of wild males provided continuous access to yeast hydrolysate, whereas sterile males deprived of access to yeast hydrolysate had much lower mating probability. Unlike males, access to yeast hydrolysate for 48 h did not increase mating probability of sterile female Q‐flies. We instead found that wild females provided continuous access to yeast hydrolysate had higher mating probability than sterile females that did or did not have 48‐h access to yeast hydrolysate. This result raises the possibility that a bisexual Q‐fly strain might operate essentially as a male‐only release when the flies are given access to yeast hydrolysate during a 48‐h pre‐release holding period. Sterile males given access to yeast hydrolysate for 48 h mated significantly earlier in the evening than wild males and, as in other recent studies, this tendency was associated with an increased tendency to mate on the trees rather than the cage walls. There was no evidence of sexual isolation in this study, as wild and sterile mass‐reared flies showed no evidence of preferential mating with their own kind. Further studies are now needed to assess the potential for pre‐release access to yeast hydrolysate to improve sexual performance and longevity of sterile, mass‐reared, Q‐flies in the field.     

Demography of a genetic sexing strain of Anastrepha ludens (Diptera: Tephritidae): effects of selection based on mating performance

相似文献   

Sexual competitiveness of sterile Ceratitis capitata males exposed to essential oils from non-host plant species native to Argentina

The sterile insect technique (SIT) is used to control fruit fly pests, such as the Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Enhancing sexual competitiveness of mass-reared sterile males can contribute to making this technique more effective. It has been shown that exposure to volatiles from essential oils (EOs), such as ginger root oil (GRO) and those from host fruits, increases male mating success. We evaluated the effect of EOs from non-host species native to Argentina, Schinus polygama (Cav.) Cabrera (Anacardiaceae) and Baccharis spartioides (Hook. & Arn.) Remy (Asteraceae), on the sexual competitiveness of sterile C. capitata males. In field cage experiments, sterile males exposed to S. polygama EO increased their mating success. In addition, sterile males exposed to this EO achieved more matings on trees than non-exposed males, thus suggesting the former are more prone to locate and defend the pheromone-calling territory. Deprivation of water and/or food significantly reduced males' life span, but exposure to S. polygama EO and GRO did not affect their survival. Schinus polygama EO is composed of mono- and sesquiterpenes with behavioral and electroantennographic responses in medflies, indicating that semiochemicals that boost medfly sexual competitiveness combine in this EO.     

Aromatherapy on a large scale: exposing entire adult holding rooms to ginger root oil increases the mating competitiveness of sterile males of the Mediterranean fruit fly in field cage trials

The sterile insect technique (SIT) is widely used in integrated programs against fruit fly pests, particularly the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Unfortunately, the mass-rearing procedures inherent to the SIT often lead to a reduction in male mating competitiveness. One potential solution involves the pre-release exposure of males to specific attractants. In particular, male exposure to ginger root oil [Zingiber officinale Roscoe (Zingiberaceae); hereafter GRO] has been shown to increase mating success dramatically in field cage trials. Initial studies exposed small groups of males (25 individuals), but more recent work has demonstrated that GRO exposure involving standard storage boxes (containing ≈ 36 000 males) also results in enhanced mating performance. The objective of the present study was to determine whether aromatization of entire trailers, holding ≈ 14 million sterile males from a genetic sexing [temperature sensitive lethal (tsl)] strain, increases male mating success. Independent of the total dose, spatial distribution, or type of dispenser used, sterile males exposed to GRO for a 24-h period displayed greater mating success than non-exposed males in mating cage trials (in which tsl males competed against males from a standard, bisexual strain for females from this same standard strain). Averaged over all experiments, tsl males exposed to GRO obtained 54% of all matings compared to 38% for non-exposed tsl males, an increase of 42%. The implications of these findings for SIT programs against C. capitata are discussed.     

Quantity and safety vs. quality and performance: conflicting interests during mass rearing and transport affect the efficiency of sterile insect technique programs

The sterile insect technique (SIT) requires production of large quantities of sterile males able to successfully compete with wild males for wild females. During eradication of a pest population, the release of fertile insects or capture of non‐marked released flies can have deleterious effects and trigger costly control measures. These perceived risks encourage program managers to apply high radiation doses and high doses of marking dye. In addition, mass rearing factories are strategically located away from release areas to prevent escape of fertile individuals within eradicated areas, raising the need for lengthy transport. Such is the case for Anastrepha obliqua Macquart (Diptera: Tephritidae) released in mango producing areas of Mexico under an SIT‐based eradication campaign. Here, we examined several standard quality‐control parameters for mass‐reared A. obliqua subjected to various time periods under hypoxia during transport, marked with different doses of fluorescent dye, and subjected to different radiation doses. Such factors were evaluated in isolation and in conjunction. Overall, long periods of hypoxia, high marking doses, and high radiation doses reduced the number of flying adults and increased the number of non‐emerged pupae. Some quality‐control parameters such as number of deformed adults, part‐emerged pupae, and non‐flying adults provided less informative guidance or redundant information of fly performance. Some tests such as mortality under stress and mating propensity in small cages were useless in detecting differences in quality among treatments for parameters evaluated during experiments. We discuss the quantity/safety‐quality/performance conflict during eradication using SIT, propose different strategies according to different stages during eradication (management, suppression, eradication, outbreaks in free areas), where males irradiated at low doses and marked with low doses of dye can be released during early suppression, and examine the pertinence of carrying out different quality‐control tests.     

Effect of diet on survival,in the laboratory and the field,of sterile male Mediterranean fruit flies

To improve the effectiveness of the sterile insect technique against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), our objectives in this study were two‐fold. First, to evaluate the ability of sterile males of the Vienna‐8 strain to survive starvation, we compared them to wild males under laboratory conditions. The second objective was to determine the effect of protein‐rich nutrition on sterile male fly survival, under starvation conditions in the laboratory, under semi‐natural conditions in a field enclosure, and under natural conditions in the open field. Therefore, we released marked sterile flies of the two diet regimes, protein‐fed or protein‐deprived, and monitored their survival by recapturing them after 4, 6, and 7 days. In the laboratory, wild males endured starvation significantly better than sterile ones and protein addition to sterile fly diet resulted in even greater reduced capability to endure starvation. On the other hand, the addition of protein to sterile‐male diet did not affect their ability to survive in a field enclosure or in the open field. We conclude that under natural conditions, where food is available, sterile male fly survival is unaffected by protein‐rich pre‐release diet.     

Incipient speciation in the Anastrepha fraterculus cryptic species complex: reproductive compatibility between A. sp.1 aff. fraterculus and A. sp.3 aff. fraterculus

Reproductive isolation among biological strains can be detected by analyzing reproductive barriers between populations. The Anastrepha fraterculus Wiedemann (Diptera: Tephritidae) cryptic species complex comprises eight morphotypes defined by morphometric characteristics. The present study analyzed the existence of pre‐ and post‐zygotic isolation among populations of A. sp.1 aff. fraterculus (A. sp.1) and A. sp.3 aff. fraterculus (A. sp.3) determined by tests of sexual compatibility, analyses of the temporal pattern of calling behavior, and the reproductive performance of the offspring of homotypic and heterotypic crosses. Pre‐zygotic tests indicated asymmetric matings with preferences for homotypic crosses. The A. sp.3 population demonstrated distinct characteristics, including low copulation percentages and a high proportion of homotypic copulations under crowded conditions. The females of the A. sp.1 population demonstrated lower copulation latency. The analyzed morphotypes demonstrated two peaks in pheromone emission activity, at the beginning and at the end of the morning and were differentiated in the abundance of males. The calling behavior of the hybrids between A. sp.1 male and A. sp.3 female crosses had an intermediate pattern compared to either parental type. Post‐zygotic compatibility tests indicated low viability of the cross between A. sp.3 males and A. sp.1 females. Pronounced asymmetries were found in the sex ratios of the offspring of heterotypic crosses, and only hybrids from A. sp.1 male vs. A. sp.3 female crosses produced descendants. The observed pre‐ and post‐zygotic incompatibilities reflect the probable occurrence of incipient speciation between A. sp.1 and A. sp.3. A more adequate taxonomic classification of this species complex that considers the distinct characteristics of each morphotype will be needed to improve environmentally wise control methods against this insect pest.     

Methoprene modulates the effect of diet on male melon fly,Bactrocera cucurbitae,performance at mating aggregations

The effect of access to dietary protein (P) (hydrolyzed yeast) and/or treatment with a juvenile hormone analogue, methoprene (M), (in addition to sugar and water) on male aggregation (lekking) behaviour and mating success was studied in a laboratory strain of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae). Six‐day‐old males were treated with (1) protein and methoprene (M+P+), (2) only protein (M?P+), or (3) only methoprene (M+P?), and compared with 14‐day‐old sexually mature untreated males (M?P?). The lekking behaviour of the four groups of males when competing for virgin sexually mature females (14 –16 days old) was observed in field cages. The following parameters were measured at male aggregations: lek initiation, lek participation, males calling, male–male interaction, female acceptance index, and mating success. For all these parameters, the M+P+ males significantly outperformed the other males. Moreover, for all parameters, there was a similar trend with M+P+ > M?P+ > M?P? > M+P?. More M+P+ males called and initiated and participated in lek activities than all other types of male, which resulted in higher mating success. They had also fewer unsuccessful copulation attempts than their counterparts. Whereas treatment with methoprene alone had a negative effect in young males with only access to sugar, access to dietary protein alone significantly improved young male sexual performance; moreover, the provision of methoprene together with protein had a synergistic effect, improving further male performance at leks. The results are of great relevance for enhancing the application of the sterile insect technique (SIT) against this pest species. The fact that access to dietary protein and treatment of sterile males with methoprene improves mating success means that SIT cost‐effectiveness is increased, as more released males survive to sexual maturity.     

橘小实蝇遗传性别品系的建立及雄性不育技术

为了更好地应用昆虫不育技术防治橘小实蝇,建立了以蛹色区分雌雄性别的橘小实蝇遗传性别品系。经过连续测交表明遗传品系能稳定遗传。质量监测表明遗传性别品系平均孵化率、蛹重、羽化率、飞出率、存活率等指标与普通种无显著差异。取遗传性别品系羽化前1、2、3天（分别用-1 d、-2 d、-3 d表示）的雄蛹,经过100 Gy的⁶⁰Co辐射后,分别与野生雌虫杂交,F1代卵的孵化率均大大低于对照组。对经过辐射处理-1 d、-2 d、-3 d的雄蛹分别进行质量检测,飞出率、存活率与对照组均无显著差异;-1 d、-2 d的羽化率之间及与对照组之间无显著差异,-3 d的羽化率与-1 d、-2 d及对照组差异均显著。取遗传性别品系的-2d的雄蛹辐射100 Gy的⁶⁰Co,获得不育雄虫,和野生雄虫一起竞争与野生雌虫交配的机会,相对不育系数为0.4923,表明不育雄虫与野生雄虫交配能力相当。田间扩散能力研究表明,不育雄虫在田间存在丰富的寄主植物的情况下不会大量扩散很远的距离。     

The sterile insect technique and the Mediterranean fruit fly: assessing the utility of aromatherapy in large field enclosures

The sterile insect technique (SIT) is widely used in integrated programs against tephritid fruit flies, particularly the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae). Unfortunately, the mass‐rearing procedures inherent to the SIT often lead to a reduction in the male mating competitiveness. One potential solution involves the prerelease exposure of males to particular attractants. In particular, male exposure to ginger root oil [Zingiber officinale Roscoe (Zingiberaceae); hereafter GRO], has been shown to increase mating success dramatically in field cage trials. To evaluate more rigorously the effectiveness of GRO exposure, we here describe two projects that compared levels of egg sterility or pupal yield, respectively, following the release of wild flies and either GRO‐exposed (treated) sterile males or GRO‐deprived (control) sterile males in large field enclosures. In both projects, sterile males from a genetic sexing strain were exposed as adults to GRO for 24 h while held in large storage boxes. In Hawaii, we dissected eggs from fruits to determine the percentage of egg hatch at four overflooding ratios, ranging from 5 : 1 to 60 : 1 (sterile : wild males), and found that, at all four ratios, the proportion of unhatched (sterile) eggs was significantly greater in enclosures containing GRO‐exposed males than control males. In Guatemala, we allowed larvae to develop in fruits and counted the number of pupae produced. At the only overflooding ratio tested (25 : 1), pupal yield was approximately 25% lower for enclosures containing GRO‐exposed males than control males, although this difference was not statistically significant. An explanation for the differing outcomes is proposed, and the implications of these findings for the SIT are discussed.     

Manipulation of insect gut microbiota towards the improvement of Bactrocera oleae artificial rearing

Bactrocera oleae (Rossi) (Diptera: Tephritidae) is the main pest of olive trees (Olea europaea L.), causing major damages in olive crops. Improvement of mass rearing is a prerequisite for the successful development of large-scale sterile insect technique (SIT) applications. This can be achieved through the enrichment of artificial diets with gut bacteria isolates. We assessed the efficiency of three gut bacteria previously isolated from Ceratitis capitata (Wiedemann), and four isolated from B. oleae, as larval diet additives in both live and inactivated/dead forms. Our results showed that dead Enterobacter sp. AA26 increased pupal weight, whereas both live and dead cells increased pupal and adult production and reduced immature developmental time, indicating that its bacterial cells serve as a direct nutrient source. Live Providencia sp. AA31 improved pupal and adult production, enhanced male survival under stress conditions, and delayed immature development. Dead Providencia sp. AA31, however, did not affect production rates, indicating that live bacteria can colonize the insect gut and biosynthesize nutrients essential for larval development. Live and dead Bacillus sp. 139 increased pupal weight, accelerated immature development, and increased adult survival under stress. Moreover, live Bacillus sp. 139 improved adult production, indicating that Bacillus cells are a direct source of nutrients. Dead Serratia sp. 49 increased pupal and adult production and decreased male survival under stress conditions whereas live cells decreased insect production, indicating that the live strain is entomopathogenic, but its dead cells can be utilized as nutrient source. Klebsiella oxytoca, Enterobacter sp. 23, and Providencia sp. 22 decreased pupal and subsequent adult production and were harmful for B. oleae. Our findings indicate that dead Enterobacter sp. AA26 is the most promising bacterial isolate for the improvement of B. oleae mass rearing in support of future SIT or related population suppression programs.