Pre-mating isolation is determined by larval rearing substrates in cactophilicDrosophila mojavensis. II. Effects of larval substrates on time to copulation,mate choice and mating propensity

Summary It has been hypothesized that reproductive character displacement has evolved in mainland Sonora, Mexico populations of cactophilicD. mojavensis due to the presence of a sympatric sibling speciesD. arizonae. In laboratory tests using ancestral Baja California populations and derived, sympatric mainland populations, asymmetrical sexual isolation has been observed among populations ofD. mojavensis where mainland females discriminate against Baja males. Effects of different pre-adult rearing environments on adult mating behaviour were assessed by comparing fermenting cactus tissues like those used in nature for breeding with laboratory media because previous studies have employed synthetic growth media for fly growth and development. Significant behavioural isolation was evident in all cases when larvae were reared on laboratory food, but was non-significant when flies were reared on fermenting cactus, except for the cactus used by most mainland populations, consistent with previous studies. Time to copulation of Baja females was greater than mainland females over all substrates, but male time to copulation did not differ between populations. Time to copulation for both sexes was significantly greater when flies were reared on laboratory food with one exception. The degree of behavioural isolation was weakly correlated with time to copulation across food types (Spearman rank correlation = 0.58,p = 0.099). Therefore, use of laboratory media in this and previous studies exaggerated adult pre-mating isolation and time to copulation in comparison to natural breeding substrates. These experiments suggest that a change in host substrates by saprophagous insects (where chemical differences exist between hosts) may have subtle effects on mating behaviour in a manner which promotes low levels of sexual isolation as a by-product of their utilization of a particular substrate during larval development. ForD. mojavensis, these results suggest that over evolutionary time, radiation into a new environment (from Baja to the mainland) allowed utilization of new host plants that may have incidentally promoted the sexual isolation patterns that have been observed within this species.See Etges (1992) for the first paper in this series.     

Entomopathogenic fungi associated with Ixodes ricinus ticks

The objective of this study was to demonstrate the occurrence of entomopathogenic fungi on Ixodes ricinus ticks in relation to the tick stage, engorgement and season. Ticks were collected from the vegetation, from small rodents and from deer. All entomopathogenic fungi found belonged to the Hyphomycetes. Paecilomyces farinosus and Verticillium lecanii were the predominant species. Other species, found only on engorged females were: Beauveria bassiana, B. brongniartii, P. fumosoroseus and V. aranearum. Eight out of 1833 ticks collected from the vegetation and three out of 269 engorged nymphs were infected with fungi. Thirty-three out of 149 engorged females were infected, whereas males and engorged larvae were not infected. Throughout the season, a significantly higher proportion of ticks collected in autumn were infected. Entomopathogenic fungi may have a significant impact on the size of the I. ricinus population, since females were the most frequently infected stage.     

Methyl 3-chloro-4-methoxybenzoate, a new candidate semiochemical inhibiting copulation behaviour of Ixodes ricinus (L.) males

Abstract.  Females of Ixodes ricinus (L. 1758) emit several semiochemicals influencing copulatory behaviour in males. One of these candidate compounds, methyl 3-chloro-4-methoxybenzoate, is emitted exclusively by engorged females, and shows a copulation inhibiting effect in males. In Y-tube experiments, the synthetic analogue of this compound at a concentration of 100 µg/mL in ethanol, attracted males in only 25.6% of tests, whereas it repelled them in 74.4% of cases. However, unengorged females do not respond to this substance at all. The number of contacts between males and unengorged females treated with methyl 3-chloro-4-methoxybenzoate (250 µg/mL in ethanol, 1 µL per female) decreases to 28% compared to a control level of 80%, and the percentage of copulating pairs decreases to 12% compared to control (49%) over 2 h. Possibly, a fertilized engorged female with mature eggs uses this semiochemical to send a signal to males indicating that another copulation is no longer needed.     

A hidden beneficial: biology of the tick‐wasp Ixodiphagus hookeri in Germany

The parasitic wasp Ixodiphagus hookeri parasitizes hard ticks and is therefore considered as a potential candidate for biological control of ticks. However, there are still considerable gaps of knowledge about the biology of I. hookeri, especially for European populations. Thus, the present study was performed to assess important life‐history parameters of the parasitoid in Germany. Field studies accomplished in three successive years revealed that unfed Ixodes ricinus nymphs are infested by the parasitoid at a low but constant rate of 1.9–3.8% and that adult wasps are present only during a short period in late summer. The mean developmental time of wasps in I. ricinus nymphs ranged from 28 to 70 days under constant laboratory conditions and was prolonged in the second half of the year. Bioassays on parasitization and host preference behaviour showed that unfed nymphs of the host species I. ricinus are significantly preferred in experiments, in which unfed and engorged larvae as well as fully engorged nymphs were offered as alternatives. The marsh tick Dermacentor reticulatus was not accepted as an alternative host. Our data show that the investigated I. hookeri populations differ markedly from populations in other regions of the world in many aspects. The adaptation of different strains to local conditions explains the limited success of imported strains in earlier biological control attempts and highlights the importance of doing research to enhance the control potential of native strains.     

Life history of the tick parasitoid Ixodiphagus hookeri (Hymenoptera: Encyrtidae) in Kenya

We conducted laboratory and field experiments to elucidate the life history of Ixodiphagus hookeri, a parasitoid of the ixodid tick Amblyomma variegatum in Western Kenya. Ixodiphagus hookeri females oviposited in unfed host nymphs as well as engorged nymphs, but rarely in engorged larvae. While I. hookeri developed to adults in engorged nymphs, the eggs laid in unfed nymphs disappeared within 2 days after oviposition. As temperature increased, development time of I. hookeri from oviposition to adult emergence in engorged nymphs decreased from 46 days at 23 °C to 35 days at 28 °C, and their immature survival in engorged nymphs decreased from 67% at 23 °C to 22% at 28 °C. No parasitoid adult emerged from hosts at 30 °C. Individual hosts parasitized by single females produced 42–53 adult wasps, 73% of which were females. As a typical pro-ovigenic species, I. hookeri females had an average of 84 mature eggs at emergence and lived only for a few days. When laboratory-reared, unfed nymphs of A. variegatum were attached to cattle for 4–9 days in subsistence farmers’ fields in Western Kenya, 25% of the engorged nymphs and 4% of the unfed nymphs on cattle were parasitized by I. hookeri, demonstrating that I. hookeri females search for and oviposit in A. variegatum nymphs on cattle. Unlike other strains of I. hookeri that overwinter as eggs in unfed nymphs, I. hookeri could continuously reproduce throughout the year in Western Kenya.     

Males prefer virgin females,even if parasitized,in the terrestrial isopod Armadillidium vulgare

In many species, males increase their reproductive success by choosing high‐quality females. In natural populations, they interact with both virgin and mated females, which can store sperm in their spermatheca. Therefore, males elaborate strategies to avoid sperm competition. In the terrestrial isopod Armadillidium vulgare, females can store sperm and produce several clutches. Moreover, this species can be parasitized by Wolbachia, which feminizes genetic males, transforming them into functional females. Our study compared attractiveness and mate choice when a male is exposed to both virgin and experienced females (i.e., females who have produced offspring and rested for 6 months), with or without Wolbachia. Our results revealed that males are more attracted to virgin females than experienced females, even if these virgin females are parasitized. Moreover, the chemical analysis highlighted different odors in females according to their reproductive and infection (Wolbachia‐free or vertically Wolbachia‐infected) status. Males attempted copulation more frequently and for longer with virgin females, even if Wolbachia‐infected, while experienced females refused further copulation. The evolutionary consequences of both male choice and female resistance on their fitness are discussed in this study.     

Genetic variation of ticks (Ixodes ricinus L.) in the Lithuanian and Norwegian populations

RAPD markers were used to measure the genetic diversity of 119 individuals of Ixodes ricinus collected from Lithuania and Norway. The samples were analysed within and also between the populations. We analysed 74 loci in each of 6 populations. Our results show high levels of diversity within the populations. The percentage of polymorphic loci of the six analysed populations: Birzai, Vilnius, Kretinga, Tjore, Kjosvik and Odderoya were 68.9%, 58.1%, 78.38%, 62.2%, 44.6% and 68.9%, respectively. The percentage of polymorphic loci in the Lithuanian populations was 93.2%, and in the Norwegian populations 81.08%. The genetic distance ranged from 0.019 to 0.079 within Norwegian populations and from 0.005 to 0.0967 within Lithuanian populations and between the countries from 0.022 to 0.146. The genetic variation of I. ricinus among Norwegian populations was lower than among Lithuanian populations. The highest part of genetic variation in I. ricinus ticks depends on variation within Kretinga (Lithuania) and Odderoya (Norway) populations situated in coastal areas where many migratory and sea birds are aggregated.     

Effets d'un régulateur de croissance d'insectes,le fénoxycarbe,sur la diapause du psylle du poirier Cacopsylla pyri

The nature of the responses of the sheep blowfly Lucilia sericata Meigen (Diptera: Calliphoridae) to carrion odours and carbon dioxide were examined in a windtunnel in the laboratory and the attractiveness of carrion-bait was examined in the field. In the wind tunnel, gravid 9-day-old females increased both the number and duration of their flights, in response to carrion odours. In addition, linear velocity was reduced and the sinuosity of flight increased in the presence of these odours. With 3-day-old females, the response to carrrion odour depended on previous exposure to proteinaceous material. Protein deprived females showed kinetic responses to carrionodour which were similar to those of gravid females. No such changes were observed in protein-fed females. No responses to CO₂ were observed. In the field, the importance of olfactory cues in bait location was demonstrated by the absence of L. sericata from sticky targets lacking an odour bait. Increased bait concentration did not affect the age, sex ratio or ratio of L. sericata to other Lucilia spp. caught, but did increase the numbers caught. Analysis of the reproductive status of females caught, indicated that a greater number of gravid females were caught than expected, while a lower number of females in the final stages of vitellogenesis were caught than expected. The results show that the responses of L. sericata to odours are complex, and are dependent on both exogenous and endogenous stimuli, the latter including the stage of ovarian development.     

Seasonal variation of vocal behaviour in a temperate songbird: assessing the effects of laboratory housing on wild-caught, seasonally breeding birds

Many laboratories are conducting research using songbirds as their animal model. In particular, songbirds are widely used for studying the behavioural and neural mechanisms underlying vocal learning. Many researchers use wild-caught birds to conduct this research, although few studies of behaviour have been conducted to determine the effects of captive housing on these species. We investigated the vocal production pattern of wild-caught black-capped chickadees (Poecile atricapillus) over an entire season in laboratory housing. We documented the frequency of production of four vocalizations (fee-bee song, chick-a-dee calls, dee calls, and gargle calls) across seasons and diurnal pattern and compared the observed pattern of laboratory vocalizations to those previously observed and reported in the wild. Laboratory-housed chickadees had seasonal and diurnal vocal production shifts that were related to both photoperiodic changes (season) and diurnal pattern. For instance, there was significantly more fee-bee song in the spring than summer, autumn, and winter with the most fee-bee song occurring at spring dawn as seen in the wild. Our results also confirmed that the general pattern of vocalizations was consistent between wild and laboratory populations, with no significant differences for either the seasonal or diurnal pattern of fee-bee song production between populations. Differences between settings were observed in the pattern of chick-a-dee calls at dawn and sunset between field and laboratory populations. However, differences in the quantity of vocalization types between laboratory and wild populations suggest that housing conditions are influencing the normal vocal behavioural patterns.     

An experimental investigation into the effects of flooding on the ixodid tick Boophilus microplus (Canestrini)

Summary The effect of water immersion on the non-parasitic stages of Boophilus microplus were investigated both under laboratory and field conditions. Survival of engorged female ticks was influenced by their age and the temperature of the water. Eggs and larvae were more resistant to submerison than were engorged females and their survival was increased at low temperatures and in water with high oxygen content. The persistence of this tick in areas prone to flooding was attributed to the survival of parasitic stages and to larvae on the pasture. Heavy rains produce favourable pasture conditions for tick reproduction so that a large increase in population size may be expected to follow such rain.     

The influence of conspecific chemical cues on walking behavior of <Emphasis Type="Italic">Ixodes ricinus</Emphasis> males

The influence of female and male cues on the male searching behavior of the sheep tick, Ixodes ricinus (Acari: Ixodidae), was studied under constant laboratory conditions by means of a computerized video tracking system. Males were released into an open glass arena in which one half had been exposed for 1 h to 5 freely walking unfed females or five males. The obtained track data were analyzed to reveal if male walking pattern was random or affected by cues of conspecifics left on the arena surface. The results showed that males spent significantly more time and walked a significantly longer distance in the area with female cues compared to the control area. Moreover, the walking path of males was more tortuous on the cues half than the control half. In contrast, walking behavior of males was not affected by male cues in any observed parameter. These results provide the first evidence that unfed females leave sex-specific semiochemicals on a substrate during their walk and that these cues have an effect on male searching behavior. Since this effect was not observed when female cues were 24-h old, the semiochemicals seem to either evaporate or are subject to degradation. The biological significance of these findings for mate-searching in I. ricinus males is discussed.     

Inter‐population mating success in Australian dengue vector mosquitoes: effects of laboratory colonization and implications for the spread of transgenics

Variability between Aedes aegypti populations in north Queensland, Australia, has the potential to impact the successful implementation of new population replacement mosquito releases for dengue control. Four Ae. aegypti colonies originating from different locations (Cairns, Mareeba, Innisfail, and Charters Towers), along with one F1 field‐derived population from Cairns, were inter‐crossed to determine any incompatibilities in copulation, insemination, and production of viable offspring. Greater copulation and insemination rates were observed when males recently introduced from the wild (‘Cairns‐Wild’ population) were mated with long‐term laboratory females. Egg viability rates for all crosses ranged from 90.2–98.2%, with no significant differences observed between crosses. Greater egg production was seen in some populations, and when corrected for wing‐length, egg production was greatest in a Mareeba x Innisfail cross (19.55 eggs/mm wing length) and lowest for the Charters Towers intra‐population cross (14.35 eggs/mm). Additionally, behavioral differences were observed between laboratory and wild mosquitoes from the Cairns location, suggesting possible laboratory conditioning. Finally, despite controlled larval rearing conditions, size differences between populations existed with Charters Towers mosquitoes consistently smaller than the other populations. The spread of genes or bacterial symbionts between these populations is unlikely to be hindered by pre‐existing reproductive barriers.     

Biology of <Emphasis Type="Italic">Dermacentor silvarum</Emphasis> (Acari: Ixodidae) under laboratory conditions

The minimum life cycle of Dermacentor silvarum Olenev had a mean duration of 87.5 days (range 74–102 days) under laboratory conditions [(27±1 °C), 70% RH, 6 L: 18 D]. The mean time in (days) for the different stages of its cycle was as follows: incubation period of eggs was 15.3 days; prefeeding, feeding and premoulting periods of larvae and nymphs averaged 5.5, 4.0 and 7.3 days, and 5.2, 5.0 and 14.6 days, respectively; prefeeding, feeding, preoviposition and oviposition periods of female adults lasted for 7.8, 4.5, 4.3 and 14.0 days, respectively. There existed a highly significant correlation between engorged body weight of females and egg masses laid (r = 0.9877, p<0.001). The reproductive efficiency index (REI) and reproductive fitness index (RFI) in females were 11.09 and 9.58, respectively. No relationship between nymphal engorged body weight and resultant sexes was observed. Delayed feeding and non-oviposition (in June and July) existed in females, and low temperature (−10 °C) treatment for 45 days could terminate oviposition diapause. However, the egg masses laid by post-diapause females were significantly smaller than those laid by females engorged in March, April and May.     

Intrasexual copulation and mate discrimination in a population of Nodilittorina radiata (Gastropoda: Littorinidae)

Intrasexual copulation and mate discrimination by Nodilittorina radiata (Gastropoda: Littorinidae) were studied on a concrete breakwater at Hakodate Bay, southern Hokkaido, Japan. Intrasexual (male–male) copulations were observed in 4.7–21.1% of copulating pairs on the shore. As females were relatively larger than males and males copulated with females larger than themselves, we hypothesized that males choose potential mates larger than themselves. However, two male mates showed no significant size preference in intrasexual copulations, suggesting that males do not choose relatively larger individuals as mates. In a laboratory mate-choice experiment, male N. radiata preferred to mate with females, indicating precopulatory sex identification. They copulated with males, however, at the frequency of 37%, perhaps because of sex misidentification.     

INHIBITION OF FEMALE DROSOPHILA MELANOGASTER REMATING BY A SEMINAL FLUID PROTEIN (ESTERASE 6)

Recent experiments have indicated that EST 6 transferred to D. melanogaster females during copulation mediates a large decrease in their attractiveness within the first six hours after mating. This decrease in attractiveness could increase the male's fitness if it were associated with less frequent remating by their mates, because a large proportion of the first male's sperm is lost if a female remates within one day. Here, I confirm a strong EST 6 influence on the receptivity of mated females; females with EST 6⁺ first mates remate significantly less frequently between 6 and 18 hours after mating than females that have not received EST 6 in the ejaculate. However, the previously observed effect of EST 6 on the attractiveness of mated females could not be substantiated. Transfer of EST 6 did not lead to a significant difference in attractiveness between EST 6°-and EST 6⁺-mated females at any time within the first 10 hours after mating for females from either of two strains.     

Sexual initiation in wild mountain gorillas

A field study of wild mountain gorillas was conducted to elucidate details of sexual initiation required for an interpretation of the increased frequency of copulation during the menstrual cycle, in comparison to the wild, that occurs when this species is tested in traditional laboratory pair tests. Although females in the wild played a clear role in establishing and maintaining proximity to the male and assertively presenting for copulation, all female presenting was preceded by some form of behavior by the male. Of particular relevance were several components of the male’s chest-beat display that had been observed at much higher frequencies in the laboratory tests. Although originally described as male aggression in the laboratory tests, such behavior now appears to be male sexual initiative, elicited by female proximity. The exaggerated form and frequency of the display in the laboratory, and the increased copulation during the cycle that ensued, are likely due to (1) placing females in proximity to a male daily, rather than on 2–3 days of estrus as occurs in the wild, and (2) the inability of the female in the laboratory to withdraw from male proximity during pair tests in a single cage. In addition to clarifying the data on laboratory pair tests, the data on wild gorillas have relevance to the captive breeding of gorillas.     

Life-history traits of the lizard Sceloporus undulatus from two populations raised in a common laboratory environment

Hatchling Sceloporus undulatus elongatus from Washington Co., Utah and S. u. garmani from Woods Co., Oklahoma were raised to maturity and reproduction under identical laboratory conditions with ad libitum food availability. Growth, allometry, age and size of maturity, clutch size and egg mass were compared among lab-raised cohorts from the two populations, among lab-raised and field-caught animals (including their field-caught mothers) and, for growth, with values obtained by previously published field studies on the same or nearby populations. For all traits population differences observed in previous field studies and current field samples resulted from both a plastic response to proximate environmental conditions and intrinsic (possibly genetic) difference. The most plastic traits were growth and age of maturity. Cohorts from both populations expressed the ability to mature in less than 6 months in the laboratory but only the S.u. garmani express early maturity in the field. Allometric differences generated during growth in the lab were not observed in field samples but may reflect an adaptive physiological difference. The least plastic trait was egg mass. The only trait for which the rank order of the difference in the field was reversed in the lab was growth rate. S.u. elongatus grew significantly faster than S.u. garmani in the lab but much slower in the field. The tendency of S.u. garmani females to breed at minimum size of maturity may be greater than that of S.u. elongatus.     

Borrelia burgdorferi sensu lato in ticks (Acari: Ixodidae) from two different foci in Spain

The prevalence of Borrelia burgdorferi sensu lato in several tick species was studied over a 2 year period in two ecologically different areas in Spain. One area was an endemic area for Lyme disease, with a number of autochthonous human cases and supported large populations of Ixodes ricinus on cattle and birds; the second area was characterized by the absence of I. ricinus together with the presence of foxes and their associated tick species. While I. ricinus was the main vector of B. burgdoreri in the endemic area (with a mean prevalence of 14% in adults and 51% in nymphs), adults of both Ixodes canisuga and Ixodes hexagonus had high rates of B. burgdorferi prevalence (30 and 28%, respectively) in the zone where I. ricinus was absent. Immatures of Ixodes frontalis were found to be carriers of the spirochete only in those zones where I. ricinus is present, suggesting evidence for reservoir competence in a tick-bird cycle.     

Toxic effect of endocytobionts fromDermacentor reticulatus ticks in three tick species after in vivo application

The endocytobionts from ovaries of partially engorged femaleDermacentor reticulatus ticks, inoculated intracoelomally into females of three tick species,D. reticulatus, Ixodes ricinus andHaemaphysalis inermis, caused considerable morphological alterations in the examined cells and tissues of the synganglion, fat body, tracheal complex and ovary of these recipients.     

Development and biological characteristics of <Emphasis Type="Italic">Haemaphysalis longicornis</Emphasis> (Acari: Ixodidae) under field conditions

The development and biological characteristics of Haemaphysalis longicornis were investigated under field conditions in Xiaowutai National Natural Reserve Area, North China. Unfed larvae, nymphs and adults were fed on rabbits and exposed to daylight. Three free-living stages were allowed to develop in field plot selected in a tick natural habitat. The host seeking behavior and seasonal occurrence were observed. Haemaphysalis longicornis were active from mid March to mid October. The premoult period of nymphs and preoviposition of females were regulating phases of their life cycle. The developmental durations of eggs, larvae and adults were constant under field conditions regardless when the development started. The oviposition periods in May and June were statistically shorter than those in July and August. The daily oviposition patterns of females engorged in May and June demonstrated unobvious peak, which differed from those engorged in July and August. The daily oviposition peak of the latter occurred on the 4th day of oviposition. Moreover, a positive correlation was found between the mass of the laid egg and the body weight of engorged females (r = 0.62, P < 0.001). The female reproductive efficiency indices were 2.9, 6.1, 10.5 and 9.0 in May, June, July and August, respectively. The mean weight (3.33 mg) of engorged nymphs molting to females was significantly higher than that (2.35 mg) of those molting to males (P < 0.001), but the body weights of both sexes were overlapping.