Laboratory-reared Females Research Articles

Chemical Exposure of Synthetic Pyrethroid on Deltamethrin Under the Selection Pressure over the Generations: A Reproductive Potential Study of Anopheles stephensi.

Biochemical synthetic pyrethroids, deltamethrin are presently used insecticides for the control of mosquito vector-borne diseases in worldwide. Mosquito re-emergence with diseases becoming a serious problem due to development of insecticide resistance. The comprehensive knowledge on the underlying mechanisms of resistance against deltamethrin is required for implementation of an efficient vector control programme. The assessment of the biological fitness of a mosquito strain exposed to insecticide pressure is extremely vital because it provides information on the development of resistance. In the present study, the adult stage of malaria vector, Anopheles stephensi, was designated for the study of deltamethrin resistance (F40 generations). The non-blood-fed, laboratory-reared females to sub-lethal doses of deltamethrin (0.004%, 0.005%, 0.007%, or 0.01%) exposed to every generation for up to F40. The adult mosquito susceptibility was performed by WHO standard method for evaluation. After 24h, mortality was recorded in both treated and control groups. Therefore, the biological fitness characteristics such as feeding, fecundity, hatchability, egg retention, immature duration, adult emergence, and adult life span were studied to assess the exposed deltamethrin under selection pressure as compared to the unexposed (control) population. The laboratory selection of An. stephensi exposed deltamethrin over the generations were diminished its biological fitness. Information on biological fitness including reproductive potential of mosquito strain under selection pressure against deltamethrin is incredibly necessary because it would facilitate in resistance management. Baseline information gives in this experiment will guide for future studies on the susceptibilities of wild malaria mosquito populations in India.

The effect of transfluthrin-treated jute and cotton emanator vests on human landing and fecundity of Anopheles minimus in Thailand

Complementary approaches to tackle outdoor and early evening biting mosquitoes are urgently required. Transfluthrin (TFT) is a volatile pyrethroid capable of altering mosquito feeding behaviour. This study investigated the efficacy of TFT-treated jute (TFT-J) and cotton (TFT-C) fabrics on human landing activity, feeding and fecundity of Anopheles minimus in Thailand. Jute and cotton fabrics each measuring 1,024 cm2 were impregnated with 7.34×10-4g/cm2 TFT (20%, w/v), and evaluated in a semi-field screen house system. Two collectors, wearing an untreated control or TFT-treated vests, conducted human-landing collections of released 100 laboratory-reared adult females of An. minimus from 18:00-00:00 h for 16 consecutive nights. Recaptured mosquitoes were given a blood meal for one hour with a membrane feeding system for assessment of blood feeding and fecundity. TFT-J, relative to control, significantly reduced human landings (Odds Ratio (OR) =0.27 (95% Confidence Interval (CI) [0.10-0.74], p=0.011)), however no significant reduction was observed for TFT-C (OR=0.67 [95% CI 0.24-1.82], p=0.43). Blood feeding was significantly lower among mosquitoes exposed to TFT-J (12.45% [95% CI, 2.04-22.85], p=0.029) and TFT-C (13% [95% CI, 0.99-26.84], p=0.016) relative to control. Impregnated fabrics had no effect on the mean number of egg oviposition. However, egg hatchability was reduced in TFT-J (49.5% [95% CI, 21.74-77.26], p=0.029) and TFT-C (40.2% [95% CI, 17.21-63.19], p=0.008) relative to control. TFT-J significantly reduced the landing, blood feeding, and fertility of An. minimus. Further studies are needed to evaluate different treatment methods on fabrics and their incorporation in integrated mosquito management.

Female preferences for high vocal effort in singing mice

Abstract Although vocalizations are essential to mammalian sociosexual behaviours, little is known about female preferences for male vocal displays in non-model mammalian species. Here, we characterized female preferences for the advertisement songs of male Alston’s singing mice (Scotinomys teguina). We developed procedures for inducing oestrus, using vaginal perforation as a morphological indicator of oestrus. We then broadcasted ‘high-effort’ and ‘low-effort’ songs recorded from wild-caught males to virgin laboratory-reared females, using a two-choice experiment. Our results indicate that females spent more time investigating speakers playing high-effort songs; surprisingly, this phonotactic response is independent of oestrus status. In wild singing mice, acoustic characteristics of high-effort male songs positively correlate with body condition. Our data suggest that females could use acoustic cues to select mates in good condition, thus providing preliminary support for adaptive mate choice hypotheses. More generally, our results support the hypothesis that elaborate Scotinomys song may be shaped by female choice.

Differential expression of Phlebotomus tobbi Adler, Theodor & Lourie, 1930 (Diptera: Psychodidae) genes under different environmental conditions

Phlebotomus tobbi is a widely distributed sand fly species in Turkey and is the proven vector of Leishmania infantum and several Phleboviruses. Information regarding the genetic basis of phenotypic plasticity is crucial for managing vector-borne diseases, as the changing environmental conditions have consequences for the survival of arthropods and the disease agents they transmit. However, limited data is available on the impacts of environmental conditions on the traits associated with sand fly survival, reproduction, and vectorial competence. The present study aimed to reveal the changes in the expression levels of three selected P. tobbi genes using laboratory-reared and wild-caught populations. A nervous system protein, Cacophony (PtCac), related to the life history traits of sand flies, and two sand fly salivary protein genes, PtSP32 and PtSP38, influence the infection of the vertebrate hosts, were assessed. Sand flies were maintained at 23°C and 27°C in the laboratory to evaluate the relationship between temperature and the expressed phenotypes. Field collections were carried out in three climatically distinct regions of Turkey to establish the regional differences in the gene expression levels of natural P. tobbi populations. In the laboratory, PtCac expression increased with the temperature. However, PtCac expression was negatively correlated with local temperature and humidity conditions. No differences were detected in the PtSP32 gene expression levels of both laboratory-reared and wild-caught females, but a negative correlation was observed with relative humidity in natural populations. Although the expression levels of PtSP38 did not differ among the females collected from distinct regions, a positive correlation was detected in the laboratory-reared colony. The findings indicated that changes in environmental conditions could drive the expression levels of P. tobbi genes, which influence population dynamics and the transmission risk of the disease.

Diel Periodicity in Males of the Navel Orangeworm (Lepidoptera: Pyralidae) as Revealed by Automated Camera Traps.

Navel orangeworm, Amyelois transitella (Walker), is a key pest of walnuts, pistachio, and almonds in California. Pheromone mating disruption using timed aerosol dispensers is an increasingly common management technique. Dispenser efficiency may be increased by timing releases with the active mating period of navel orangeworm. Past work found that the peak time of sexual activity for navel orangeworm females is 2 h before sunrise when temperatures are above 18°C. Inference of male responsiveness from data collected in that study was limited by the necessity of using laboratory-reared females as a source of sex pheromone emission to attract males and the inherent limitations of human observers for nocturnal events. Here we used camera traps baited with artificial pheromone to observe male navel orangeworm mating response in the field over two field seasons. Male response to synthetic pheromone exhibited diel patterns broadly similar to females, i.e., they were active for a brief period of 2–3 h before dawn under summer conditions and began responding to pheromone earlier and over a longer period of time during spring and fall. But contrary to the previous findings with females, some males were captured at all hours of the day and night, and there was no evidence of short-term change of pheromone responsiveness in response to temperature. Environmental effects on the response of navel orangeworm males to an artificial pheromone source differ in important ways from the environmental effects on female release of sex pheromone.

Electroantennographic Responses of Wild and Laboratory-Reared Females of Xyleborus affinis Eichhoff and Xyleborus ferrugineus (Fabricius) (Coleoptera: Curculionidae: Scolytinae) to Ethanol and Bark Volatiles of Three Host-Plant Species.

Simple SummaryThe ambrosia beetles Xyleborus affinis and Xyleborus ferrugineus are wood borers reported as secondary vectors of pathogenic fungi that cause lethal vascular diseases in mango, cacao, and trees within the laurel family. The use of specific attractants or repellants is one potential method for monitoring or controlling these pests. Chemical ecology studies to develop such tools often use wild or laboratory-reared beetles without first determining whether there are differences in their responses. We compared the antennal olfactory responses of wild and laboratory-reared X. affinis and X. ferrugineus to bark odors of gumbo-limbo (Bursera simaruba), mango (Mangifera indica) and chinini (Persea schiedeana) with different aging times and used GC–MS to analyze the chemical composition of these bark odors. The antennal responses of laboratory-reared and wild females differed in X. affinis and X. ferrugineus when interacting with odors. In addition, both beetle species displayed stronger antennal responses to aged bark odors of gumbo-limbo and chinini, apparently due to changes in volatile emissions over time.Chemical ecology studies on ambrosia beetles are typically conducted with either wild or laboratory-reared specimens. Unlike laboratory-reared insects, important aspects that potentially influence behavioral responses, such as age, physiological state, and prior experience are unknown in wild specimens. In this study, we compared the electroantennographic (EAG) responses of laboratory-reared and wild X. affinis and X. ferrugineus to 70% ethanol and bark odors (host kairomones) of Bursera simaruba, Mangifera indica, and Persea schiedeana aged for 2, 24, and 48 h. Chemical analyses of each odor treatment (bark species x length of aging) were performed to determine their volatilome composition. EAG responses were different between laboratory-reared and wild X. ferrugineus when exposed to ethanol, whereas wild X. affinis exhibited similar EAG responses to the laboratory-reared insects. Ethanol elicited the strongest olfactory responses in both species. Among the bark-odors, the highest responses were triggered by B. simaruba at 48 h in X. affinis, and P. schiedeana at 24 and 48 h in X. ferrugineus. Volatile profiles varied among aged bark samples; 3-carene and limonene were predominant in B. simaruba, whereas α-copaene and α-cubebene were abundant in P. schiedeana. Further studies are needed to determine the biological function of B. simaruba and P. schiedeana terpenes on X. affinis and X. ferrugineus, and their potential application for the development of effective lures.

Tenuipalpus uvae (Acari: Tenuipalpidae) and Calophya spondiadis (Hemiptera: Psyllidae), pests of Spondias in Florida, USA

Two main pests were found attacking purple mombin or Spanish plum, Spondias purpurea (Sapindales: Anacardiaceae) in Homestead, Florida, USA: Tenuipalpus uvae De Leon (Acari: Tenuipalpidae), a flat mite damaging the leaves, and the spondias psyllid, Calophya spondiadis Burkhardt & Mendez (Hemiptera: Psyllidae) feeding on the flowers, fruitlets, and tender expanding leaves. Tenuipalpus uvae populations are composed principally of females. Female longevity was 13.1 ± 2.0 d and unmated females oviposited 5.0 ± 1.0 eggs in their lifetime. On average, eclosure time for eggs from laboratory-reared unmated females was 10.2 ± 0.1 d, and developmental times for the larva, protonymph, and deutonymph were 6.6 ± 0.4 d, 5.7 ± 0.2 d, and 6.8 ± 0.7 d, respectively. In a survey of S. purpurea leaflets, the upper surface had a significantly higher number of nymph and adult mites (6.43 ± 0.60) than the lower surface (2.29 ± 0.35). Calophya spondiadis laid their eggs on flowers at first, because these were the first to sprout in Jan, and they were heavily infested by the end of Mar with an average of 10.9 ± 1.5 eggs and 9.6 ± 1.1 nymphs per flower. Few eggs or nymphs (0.17 ± 0.17 and 0.08 ± 0.08, respectively) were found per fruit; nonetheless, the fruit did have multiple punctures or necrotic spots (16.7 ± 4.3 per fruit), likely acquired as nymphs fed on the flowers and their ovaries. Significantly more eggs were found on the upper surface of the leaves and the petiole portion adjacent to each leaflet than the underside of the leaves, and most nymphs migrated to the undersides. The feeding damage resulted in black, necrotic spots that became brown pits as the leaves matured and hardened.

Microbiota Variation Across Life Stages of European Field-Caught Anopheles atroparvus and During Laboratory Colonization: New Insights for Malaria Research.

The potential use of bacteria for developing novel vector control approaches has awakened new interests in the study of the microbiota associated with vector species. To set a baseline for future malaria research, a high-throughput sequencing of the bacterial 16S ribosomal gene V3-V4 region was used to profile the microbiota associated with late-instar larvae, newly emerged females, and wild-caught females of a sylvan Anopheles atroparvus population from a former malaria transmission area of Spain. Field-acquired microbiota was then assessed in non-blood-fed laboratory-reared females from the second, sixth, and 10th generations. Diversity analyses revealed that bacterial communities varied and clustered differently according to origin with sylvan larvae and newly emerged females distributing closer to laboratory-reared females than to their field counterparts. Inter-sample variation was mostly observed throughout the different developmental stages in the sylvan population. Larvae harbored the most diverse bacterial communities; wild-caught females, the poorest. In the transition from the sylvan environment to the first time point of laboratory breeding, a significant increase in diversity was observed, although this did decline under laboratory conditions. Despite diversity differences between wild-caught and laboratory-reared females, a substantial fraction of the bacterial communities was transferred through transstadial transmission and these persisted over 10 laboratory generations. Differentially abundant bacteria were mostly identified between breeding water and late-instar larvae, and in the transition from wild-caught to laboratory-reared females from the second generation. Our findings confirmed the key role of the breeding environment in shaping the microbiota of An. atroparvus. Gram-negative bacteria governed the microbiota of An. atroparvus with the prevalence of proteobacteria. Pantoea, Thorsellia, Serratia, Asaia, and Pseudomonas dominating the microbiota associated with wild-caught females, with the latter two governing the communities of laboratory-reared females. A core microbiota was identified with Pseudomonas and Serratia being the most abundant core genera shared by all sylvan and laboratory specimens. Overall, understanding the microbiota composition of An. atroparvus and how this varies throughout the mosquito life cycle and laboratory colonization paves the way when selecting potential bacterial candidates for use in microbiota-based intervention strategies against mosquito vectors, thereby improving our knowledge of laboratory-reared An. atroparvus mosquitoes for research purposes.

Videographic analysis of flight behaviours of host-seeking Anopheles arabiensis towards BG-Malaria trap.

The BG-Malaria trap (BGM) is an adaptation of the well-known BG-Sentinel trap (BGS) with greater trapping efficiencies for anopheline and culicine mosquitoes. Its continued optimization requires greater understanding of mosquito flight behaviors near it. We used three high-resolution infrared cameras (68 frames/second) to track flight behaviors of laboratory-reared Anopheles arabiensis females in vicinity of the BGM in comparison with BGS. Additional comparisons were done for BGM at 20, 40 and 80cm heights, and for BGMs baited with Ifakara blend plus CO2, CO2 alone, or no bait. More mosquitoes were observed near BGM than BGS. Both BGMs installed 20cm above the floor and baited with CO2 received more visits by host-seeking mosquitoes than the other BGMs evaluated in their respective experiments. Trap designs, height and attractants all influence mosquito activity in vicinity of the traps which can be readily visualized using infrared cameras to accelerate trap development and testing. The greater activity of host-seeking mosquitoes near BGM than BGS supports the proven superiority of BGM traps in field and semi-field settings.

Effect of overwintering on survival and vector competence of the West Nile virus vector Culex pipiens

BackgroundWest Nile virus (WNV) is a mosquito-borne virus that is mainly transmitted among birds by Culex pipiens mosquitoes. The species Cx. pipiens consists of two biotypes named pipiens and molestus, which together can form hybrids. One of the major distinctions between the biotypes is their overwintering behaviour. Adults of biotype pipiens diapause during winter, whereas biotype molestus remains actively blood-feeding. Diapausing may affect survival and vector competence of biotype pipiens. The aims of this study were therefore to identify the biotype composition of diapausing Cx. pipiens mosquitoes, to quantify survival throughout the autumn and winter months, and to determine effects of overwintering on vector competence of emerging Cx. pipiens mosquitoes for WNV.MethodsDiapausing mosquitoes were collected at two typical overwintering locations in the Netherlands. A selection of Cx. pipiens mosquitoes was identified to biotype using real-time PCR. Survival of diapausing Cx. pipiens mosquitoes during autumn and winter was monitored by placing cages with either field-collected or laboratory-reared females in houses and sheds. Vector competence of field-collected (diapausing) and laboratory-reared (non-diapausing) Cx. pipiens mosquitoes was determined to gain insight in the effect of overwintering on WNV transmission.ResultsThe majority (92%) of diapausing Cx. pipiens females were identified as biotype pipiens. More than 70% of diapausing Cx. pipiens mosquitoes was able to survive for more than four months in sheds, whereas diapausing in houses resulted in 100% mortality in that same period. In contrast, non-diapausing Cx. pipiens biotype pipiens mosquitoes reared in the laboratory died within a week in both houses and sheds. Vector competence of Cx. pipiens mosquitoes that had diapaused during the autumn and winter months was comparable to non-diapausing laboratory-reared mosquitoes.ConclusionsThis study confirms that the majority of Cx. pipiens mosquitoes in their typical overwintering site belongs to the pipiens biotype. It shows that more than two-third of diapausing Cx. pipiens mosquitoes is able to survive winter under sheltered winter conditions. Finally, vector competence for WNV of mosquitoes that emerge from overwintering sites is not affected by their relatively old age.

Effects of γ radiation on the reproduction and enteroendocrine cells of the spruce bark beetle Ips typographus and prospects for its control

The European spruce bark beetle Ips typographus is one of the most important pests of spruce forests in Europe. The present study investigated the feasibility of a sterile insect technique (SIT) for this pest control. Laboratory-reared males were exposed to various doses of γ radiation (0, 10, 30, 50, 70 and 90 Gy) and allowed to mate with laboratory-reared untreated females. The radiation significantly affected the number of viable offspring. Although females oviposited on average 16.0–18.8 eggs per maternal tunnel in all treatments, the number of larval tunnels per maternal tunnel observed after 4 weeks of development decreased from 17.3 in the control (0 Gy) to 4.5 in the 90-Gy group. Numerous oviposition sites without larval tunnels were observed in galleries when males received high doses (70 and 90 Gy) of γ radiation. Comparison of the sperm viability in the control and irradiated males did not reveal any statistically significant differences. Side effects of the irradiation were examined by immunostaining of the enteroendocrine cells. Two distinct types of cells were revealed with antibodies to neuropeptides allatostatin A and tachykinin in the midgut. The alimentary tracts of males receiving a high dose of radiation (90 Gy) showed a significantly decreased number of tachykinin-like immunoreactive enteroendocrine cells. This observation suggests potential radiation-induced damage of the digestive system, which could lead to a reduction in male fitness. The implications of these findings for successful use of SIT in spruce bark beetle control are discussed.

Male white grub beetles prefer the pheromone composition of young females in the field.

Females of the white grub beetle, Dasylepida ishigakiensis, release both (R)- and (S)-2-butanol as sex pheromones, but the males are only attracted to (R)-2-butanol. In laboratory-reared females, the proportion of the (R)-isomer decreased significantly as their calling opportunities increased and as they aged. We examined whether such qualitative changes also occur in field populations. We collected virgin females from the field and then trapped and analysed the volatiles emitted during their first and second callings. The ratio of (R)- to (S)-2-butanol (R/S) was 78:22 at the first calling, but shifted to 39:61 at the second calling. While investigating the composition of the female pheromones, the question arose as to whether the male preferences change in response to the shift in female pheromone composition. To answer this question, we observed the behaviour of young and old males in response to various R/S ratios as lures in the laboratory and in the field. In the flight tunnel assay of laboratory-reared individuals, young males touched female models with a 9:1 R/S ratio lure less than those with pure (R)-2-butanol; however, older males touched the two groups with equivalent frequency. In the field trap test, older males were much more attracted to (R)-2-butanol-scented lures. When we tested using lures with the same amount of (R)-2-butanol but added different amounts of the (S)-isomer, we found that increased levels of (S)-2-butanol resulted in lower attractiveness to males. (S)-2-butanol was confirmed to have an inhibitive activity in the attractiveness of (R)-2-butanol.

Paternity analyses in wild-caught and laboratory-reared Caribbean cricket females reveal the influence of mating environment on post-copulatory sexual selection.

Polyandry is ubiquitous in insects and provides the conditions necessary for male- and female-driven forms of post-copulatory sexual selection to arise. Populations of Amphiacusta sanctaecrucis exhibit significant divergence in portions of the male genitalia that are inserted directly into the female reproductive tract, suggesting that males may exercise some post-copulatory control over fertilization success. We examine the potential for male-male and male-female post-copulatory interactions to influence paternity in wild-caught females of A. sanctaecrucis and contrast our findings with those obtained from females reared in a high-density laboratory environment. We find that female A. sanctaecrucis exercise control by mating multiple times (females mount males), but that male-male post-copulatory interactions may influence paternity success. Moreover, post-copulatory interactions that affect reproductive success of males are not independent of mating environment: clutches of wild-caught females exhibit higher sire diversity and lower paternity skew than clutches of laboratory-reared females. There was no strong evidence for last male precedence in either case. Most attempts at disentangling the contributions of male-male and male-female interactions towards post-copulatory sexual selection have been undertaken in a laboratory setting and may not capture the full context in which they take place--such as the relationship between premating and post-mating interactions. Our results reinforce the importance of designing studies that can capture the multifaceted nature of sexual selection for elucidating the role of post-copulatory sexual selection in driving the evolution of male and female reproductive traits, especially when different components (e.g. precopulatory and post-copulatory interactions) do not exert independent effects on reproductive outcomes.

Effect of Spring and Winter Temperatures on Winter Moth (Geometridae: Lepidoptera) Larval Eclosion in the Northeastern United States.

Field and laboratory experiments were conducted to elucidate various factors influencing the temperature-dependent larval eclosion of winter moth, Operophtera brumata L, in New England. We found no difference in duration of the embryonic stage of eggs reared from larvae collected in Massachusetts (MA) and on Vancouver Island, British Columbia (BC), where winter temperatures are rarely below freezing. The number of growing degree days (GDD) required for larval eclosion declined with the number of days chilled in the laboratory and number of days below freezing in the field, confirming the findings of previous studies. Thus, eggs hatched with fewer GDD, when the spring came later than usual. Date of oviposition had no effect on date of hatch. Eggs laid by naturally occurring (feral) females hatched sooner, with lower GDD, than eggs from laboratory-reared females from MA and BC held on the same trees over the winter. South-facing eggs on the stems of trees hatched on average 1.6 days sooner than north-facing eggs. GDD calculated from bihourly measures of temperature were 15% greater than GDD estimates based on the average of daily maximum and minimum temperatures, as used by many GDD estimates made for online sources. Over two years, the mean GDD in °C for hatch of feral eggs was 176.53 ± 6.35 SE based on bihourly temperature measurements, a 1 January start date, and a 3.9°C developmental threshold. This value varied markedly, however, between the two years.

Effect of the symbiont Candidatus Erwinia dacicola on mating success of the olive fly Bactrocera oleae (Diptera: Tephritidae)

Mutualistic bacterial endosymbionts provide many benefits to their insect hosts, but their role in mating has not been studied in the past. In this study, we examined copulatory success and mating latency as two parameters of mating success to assess the influence of Candidatus Erwinia dacicola on mating between a laboratory population of olive flies (Bactrocera oleae Rossi) of Israel origin and a wild population of olive flies from Israel. Previous studies have shown that in many species of tephritid flies, laboratory-reared males have lower fitness and achieve fewer matings than wild males. Our research has shown that this Israeli population of olive flies reared in the laboratory on an artificial diet lacked an endosymbiont, Ca. E. dacicola, found in wild-caught insects from Israel. We hypothesized that decreased fitness and mating ability in laboratory-reared flies could be due to the absence of this endosymbiont. Mating assays between both sexes of these two Israeli populations revealed matings to occur primarily between laboratory-reared females and wild males. Laboratory-reared males achieved only 22% of the total matings. Candidatus Erwinia dacicola was found in significantly fewer insects from the laboratory population than in the wild population; within populations, male and female olive flies were equally likely to have the endosymbiont. However, differences in readiness to mate between the two populations, and not the presence of the endosymbiont, explained mating latency.

Reproduction, development and habits of the large turkey louse Chelopistes meleagridis (Phthiraptera: Ischnocera) under laboratory conditions.

The bionomy of Chelopistes meleagridis off the host was observed with the aim of better understanding the aspects of this species' life cycle. For this purpose, C. meleagridis adults were collected and maintained under controlled conditions to reproduce (35°C and RH > 80%), with turkey feathers as the food source. From the offspring of these lice, the development of 150 individuals was observed from the egg to the adult phase. These eggs were divided into two groups of 75 each. After hatching, one group was given a diet composed of feathers while the other received feathers plus skin of the host turkey (Meleagris gallopavo). The "feather + skin" diet resulted in the greatest number of adults, so this diet was given to the next generation of lice reared in vitro, starting from the first instar, to observe their fertility, fecundity and longevity. High reproduction rates were found in relation to other lice of the Ischnocera sub-order, particularly the number of eggs per day and number of eggs produced per female over the lifetime (means of 2.54 and 26.61 eggs, respectively, for wild females and 2.11 and 29.33 eggs for laboratory-reared females). The inclusion of skin in the diet was a determining factor for development to the adult stage, since 48% of the lice fed this diet reached that stage, versus 1.3% that reached maturity fed only with feathers. The development time of the males and females was similar (mean of 29.38 days), without any difference in the sexual proportion of the adults.

Managing the white grub beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) in sugarcane fields on Miyako Island, Japan, using sex attractant pheromone: effects of mating delay on the reproductive ability of laboratory-reared and field-collected females

The use of pheromones to disrupt mating behaviours is perhaps the most promising method for controlling the sugarcane pest Dasylepida ishigakiensis Niijima et Kinoshita, also known as the white grub beetle. Herein, based on laboratory observations of mating, oviposition and hatching behaviours in the white grub beetle, we suggest guidelines for the timing and length of sex-pheromone release in the field. In the present study, we collected virgin female adults early in the mating season (4 February) in sugarcane fields in Okinawa, Japan and mated them in the laboratory after different time intervals with males collected at the same time. We investigated the effects of delayed mating on mating success, timing of oviposition, female longevity after mating and progeny hatchability. Mating success - defined as the proportion of mated females that produced progeny — and hatchability were not significantly affected by mating delays of less than 31 days, although further delays dramatically decreased these parameters. Female life span and the time between mating and oviposition were unaffected by mating delay. Similar results were obtained for laboratory-reared adults mated on day 45, 60 or 80 after adult emergence. Based on these findings, we propose that releasing synthetic sex pheromones between mid-January (i.e. shortly before the start of the mating season) and late March would be optimal for disrupting the mating behaviours of this beetle in the field.

A Transect for Reproductive Compatibility and Evidence for a “Hybrid Sink” in a Hybrid Zone ofHyalophora(Insecta: Lepidoptera: Saturniidae)

Egg fertility, embryo viability, and fecundity were measured, using experimental hybridization, to produce a profile of reproductive compatibility across a hybrid zone between Hyalophora euryalus (Boisduval, 1855) and H. columbia gloveri (Strecker, 1872) (Lepidoptera: Saturniidae) in the Sierra Nevada mountains of California. Three aspects of the genetic structure of the hybrid zone are discussed: 1) hybrid zone females have normal fecundity, in contrast to typically barren laboratory-reared F1 hybrid females from crosses between widely allopatric parental taxa; 2) hybrid zone females from defined sites along the transect, in comparison to allopatric species matings, showed no reduction in fertility or viability in matings with males from the same site; and 3) a “hybrid sink” was discovered, the moths from which were the least compatible among various inter-site crosses. Topography affects the genetic structure of the hybrid zone, especially by reducing gene flow in relation to rain shadows and associated ecological features. Within the hybrid zone, natural selection has apparently modified postzygotic isolation by favoring compatible genotypes regulating öogenesis. The long-term structure and stability of the hybrid zone is discussed in terms of ecological heterogeneity in complex montane landscapes. The suitability of the “tension zone” model, which depicts an equilibrium between the effects of selection balanced against dispersal, is discussed for the Hyalophora hybrid zone.

Pheromonal Divergence Between Two Strains of Spodoptera frugiperda

Spodoptera frugiperda consists of two genetically and behaviorally different strains, the corn- and the rice-strain, which seem to be in the process of sympatric speciation. We investigated the role of strain-specific sexual communication as a prezygotic mating barrier between both strains by analyzing strain-specific variation in female pheromone composition of laboratory and field strains, and also male attraction in wind tunnel and field experiments. Laboratory-reared and field-collected females from Florida exhibited strain-specific differences in their relative amount of (Z)-7-dodecenyl acetate (Z7-12:OAc) and (Z)-9-dodecenyl acetate (Z9-12:OAc). In wind tunnel assays, we did not find strain-specific attraction of males to females. However, in field experiments in Florida, we observed some differential attraction to synthetic pheromone blends. In a corn field, the corn-strain blend attracted more males of both strains than the rice-strain blend, but both blends were equally attractive in a grass field. Thus, habitat-specific volatiles seemed to influence male attraction to pheromones. In dose-response experiments, corn-strain males were more attracted to 2 % Z7-12:OAc than other doses tested, while rice-strain males were attracted to a broader range of Z7-12:OAc (2-10 %). The attraction of corn-strain males to the lowest dose of Z7-12:OAc corresponds to the production of this compound by females; corn-strain females produced significantly smaller amounts of Z7-12:OAc than rice-strain females. Although corn-strain individuals are more restricted in their production of and response to pheromones than rice-strain individuals, it seems that differences in sexual communication between corn- and rice-strain individuals are not strong enough to cause assortative mating.

Mortality and reproductive effects of ingested spinosad on adult bollworms

Upon emergence from their pupal cells, bollworm, Helicoverpa zea (Boddie), adults actively seek and feed on plant exudates before they disperse and reproduce on suitable host plants. This nocturnal behavior of the bollworm may be exploited as a pest management strategy for suppression of the insect by using an attractant/stimulant mixed with an insecticide to induce feeding to cause adult mortality or reproductive reduction/inhibition. This study aimed to determine in the laboratory whether or not spinosad when mixed with sucrose solution as a feeding stimulant and ingested by bollworm could influence mortality and reproduction of the insect. Sublethal concentrations of spinosad fed to laboratory-reared females confined with males significantly reduced percentage hatch of eggs at 0.1 mg L(-1), and it was reduced to near zero at 2.5 mg L(-1) when compared with females fed 2.5 M sucrose solutions only. The lethal concentration (LC(99)) for males captured from the field in sex-pheromone-baited traps was 73 mg L(-1) for 24 h response. Proboscis extension response was not inhibited significantly even at 10 g L(-1). In spite of a 137-fold increase in lethal dose concentration, spinosad did not inhibit feeding. A detailed study of laboratory-reared and field-collected bollworm adults relative to mortality and reproduction after ingestion of spinosad indicates that spinosad would be useful in an attract-and-kill strategy to control the insect when mixed with a feeding attractant/stimulant. Field validation of the data is warranted.