The contribution of multiple mating and spermatophore consumption to the lifetime reproductive success of female field crickets (Gryllus bimaculatus)

ABSTRACT.

• 1 The adaptive significance of multiple mating by female Gryllus bimaculatus (De Geer) was investigated.
• 2 Multiple mating prevented the depletion of sperm stores and, therefore, maintained high hatching success. This may not, however, explain the high frequency of remating in this species.
• 3 Male-derived egg stimulants known to be passed with sperm at mating increased the number of eggs produced only when females mated throughout their lifespans.
• 4 Spermatophore consumption appeared to provide nutrients which, while they did not increase the quantity of eggs, increased egg quality as indicated by weight. Females who consumed spermatophores had a greater hatching success.
• 5 While females may derive non-genetic benefits from mating, these are apparently long-term benefits; females must mate throughout their lives in order to accrue them.
• 6 Since the benefits of mating may not be derived from individual males, the spermatophores and their contents in this species are best considered as mating effort.

     

Female choice in the field cricket Gryllus bimaculatus (De Geer)

Females of the field cricket G. bimaculatus show multiple mating. Since such a strategy is likely to incur costs such as time and energy spent in orientation to calling males or increased susceptibility to predation, it must have certain selective advantages. This study shows that multiple mating is a potential mechanism of mate choice. Females removed the spermatophores of some males before complete insemination was achieved. Females also remained with large males in order to mate a second or third time. Since spermathecal morphology may limit last-male sperm precedence in this species, a function of multiple mating may be to dilute the sperm stored from previous matings with that of the current male, so increasing his representation in offspring production.     

Male size,mating potential and lifetime reproductive success in the field cricket,Gryllus bimaculatus (De Geer)

Male size is known to influence short-term, competitive mating success in the field cricket, Gryllus bimaculatus. Short-term measures of mating success are, however, potentially misleading when considering the influence of sexual selection on male morphology and behaviour. The lifetime mating success of males was investigated in the absence of competition to determine the effects of male size per se on reproductive potential and any effects of female behaviour on male lifetime mating success. The cost of spermatophore production was relatively greater for small males who consequently had a longer refractory period between matings, thus limiting their reproductive potential. Large males had a mating advantage resulting from a differential response of females to the courtship attempts of large and small males. Furthermore, small males were ‘sexually incompetent’ in terms of their ability to attach spermatophores once mounted. There was, therefore, an overall positive relationship between male size and lifetime reproductive success.     

Inter-male competition and mating success in the field cricket,Gryllus bimaculatus (de Geer)

Males of the field cricket Gryllus bimaculatus compete aggressively for the acquisition of burrows and females. The effect of population density on the nature of encounters between males, and factors affecting individual competitive ability were examined. Aggression was reduced in high population densities, both in terms of the proportion of encounters leading to aggression and in the intensity of aggressive disputes. The addition of burrows and females increased aggressiveness and removed population density effects. Individual competitive ability was determined primarily by body size and secondly, by an individual's past experience of winning (‘confidence’). Competitive ability, prior residence in burrows and the presence of females all appear to influence the outcome of aggressive disputes. Variation in calling frequency of males was influenced both by population density and competitive ability; males who called more often had a greater encounter rate with females. Females were more likely to mate with burrow residents. However, amongst residents, larger males gained more matings. The potential for female choice in this species is discussed.     

Fertilization,syngamy, and early embryonic development in the cricket Gryllus bimaculatus (De Geer)

Fertilization and early embryonic mitoses of the cricket Gryllus bimaculatus were examined by fluorescence staining of whole-mount as well as squash preparations. Egg meiosis occurs near the ventral surface of the egg, while sperm transforms into a sperm pronucleus in the cytoplasmic island on the dorsal side. After meiosis, the egg pronucleus moves across the egg toward the sperm pronucleus in the island, where union of these nuclei occurs. The first cleavage mitosis is gonomeric, as in insects such as Pyrrhocoris, Drosophila, and Bombyx. After the third cleavage the synchrony of nuclear division is lost and the dividing nuclei are distributed all over the egg by 12 h after deposition.     

Hyperacute directional hearing and phonotactic steering in the cricket (Gryllus bimaculatus deGeer)

Background

Auditory mate or prey localisation is central to the lifestyle of many animals and requires precise directional hearing. However, when the incident angle of sound approaches 0° azimuth, interaural time and intensity differences gradually vanish. This poses a demanding challenge to animals especially when interaural distances are small. To cope with these limitations imposed by the laws of acoustics, crickets employ a frequency tuned peripheral hearing system. Although this enhances auditory directionality the actual precision of directional hearing and phonotactic steering has never been studied in the behaviourally important frontal range.

Principal Findings

Here we analysed the directionality of phonotaxis in female crickets (Gryllus bimaculatus) walking on an open-loop trackball system by measuring their steering accuracy towards male calling song presented at frontal angles of incidence. Within the range of ±30°, females reliably discriminated the side of acoustic stimulation, even when the sound source deviated by only 1° from the animal''s length axis. Moreover, for angles of sound incidence between 1° and 6° the females precisely walked towards the sound source. Measuring the tympanic membrane oscillations of the front leg ears with a laser vibrometer revealed between 0° and 30° a linear increasing function of interaural amplitude differences with a slope of 0.4 dB/°. Auditory nerve recordings closely reflected these bilateral differences in afferent response latency and intensity that provide the physiological basis for precise auditory steering.

Conclusions

Our experiments demonstrate that an insect hearing system based on a frequency-tuned pressure difference receiver achieves directional hyperacuity which easily rivals best directional hearing in mammals and birds. Moreover, this directional accuracy of the cricket''s hearing system is reflected in the animal''s phonotactic motor response.     

the effect of pair formation on diel calling patterns in two cricket species,Gryllus veletis andGryllodes sigillatus (Orthoptera: Gryllidae)

After pair formation, male crickets should reduce calling to minimize the risks to males of attracting predators and/or rivals. We tested this hypothesis in two cricket species, one in which males exhibit a high mating propensity (Gryllus veletis)and another in which the mating propensity of males is constrained by the manufacture of elaborate, bipartite spermatophores (Gryllodes sigillatus).Calling durations of male G. veletisdeclined precipitously after the introduction of females but remained unchanged in G. sigillatus.We attribute the asymmetric effect of female proximity on male calling to differences in the mating propensity of males of the two species. Male G. veletisabstain from further calling in favor of repeated matings with the same female. Male G. sigillatusprobably lack the opportunity to mate repeatedly with the same female and, thus, resume calling shortly after mating to increase their probability of attracting additional mates.     

Neuroplasticity and phonotaxis in monaural adult female crickets (Gryllus bimaculatus de Geer)

Summary One foreleg was amputated at mid-femur in adultGryllus bimaculatus females. In phonotaxis tests these monaural crickets show course deviations and circling towards the intact side (Fig. 1). Mean course stability is best at 60 and 70 dB (Fig. 2). Here it differs significantly from a threshold value for orientated walking in females operated on the day of adult moult, but not in those operated two weeks later. The orientational performance improves with the interval between amputation and test (Fig. 3).Centripetal cobalt backfills reveal degeneration of tympanal nerve fibers on the amputated side (Fig. 4B, C). The mean number of intact afferents crossing the midline of the prothoracic ganglion is increased in monaural versus binaural crickets. Maximum transmidline extension is not correlated with the period of deafferentation (Fig. 5).Intracellular recording and staining of prothoracic auditory interneurons shows some axonal sprouts in ON1i (intact side) and ON2, but no significant physiological changes (Figs. 6A, D; 8A, C, E, G). Apart from axonal sprouts ON1a (amputated side) may show a few dendritic sprouts into the intact auditory neuropil (Figs. 6C, 7). Excitation in some ON1a-cells reveals functional contacts to intact auditory afferents (via crossing dendrites or possibly crossing afferents, Figs. 6e, 7, 8F). Morphological and associated physiological changes start early in AN2a (amputated side). The degree of crossing dendrites and contralateral excitation increases with postoperative age (Figs. 8H, 9).     

Detailed developmental morphology of the spermatophore of the Mediterranean field cricket, Gryllus bimaculatus (De Geer) (Orthoptera: Gryllidae)

The detailed developmental morphology of the spermatophore of a cricket (Gryllus bimaculatus, De Geer) is described for the first time. A specific behavioural reference point is identified from which time developmental processes proceed in a more or less robotic manner. The structures previously identified as forming part of the fully formed spermatophore, that is, the ampulla, the sperm tube and the attachment plate are all identified in detail during their development. Interestingly the attachment plate material is extruded as an amorphous mass from the ejaculatory duct prior to its insertion into the dorsal pouch, the sperm do not enter the sperm sac until the end of spermatophore formation, and the sperm tube itself is formed from a flattened layer of material which rolls up into a tube and involves the use of a die mechanism.     

Agonistic Displays and the Benefits of Fighting in the Field Cricket, Gryllus bimaculatus

Fighting is often composed of discrete agonistic displays. Few studies have partitioned fighting behavior into its component agonistic displays and evaluated the relationships between the frequency of the displays and the potential benefits of fighting, particularly mating success. In this study, we quantified the frequency of male field cricket, Gryllus bimaculatus, agonistic displays. The displays were quantified under three social environments which varied in the potential benefits of fighting: males with other males only, males with other males and female scents, and males with other males and females. We found that (1) the presence of females elicited an increase in agonistic displays characteristic of intermediate levels of escalation, (2) female scents did not produce a similar increase in the frequency of agonistic displays, and (3) in the presence of females, the frequency of agonistic displays was positively correlated with mating success. Aggressive stridulation, an energetically low-cost display, was more strongly associated with mating success than were more costly displays. The results are discussed in the context of the evolutionary theory of aggression and in the context of cricket mating systems.     

The effects of injecting Juvenile Hormone III into the prothoracic ganglion on phonotaxis by female crickets Gryllus bimaculatus

Nanoinjection of Juvenile Hormone III (JH III) into the prothoracic ganglion causes virgin female crickets Gryllus bimaculatus De Geer to become more phonotactically selective for the syllable periods (SPs) of model calling songs. Females responding to all, or almost all, of the SPs presented before JH III injection significantly narrow their responses to a range of SPs that is usually centered on the SPs included in the conspecific males' calling song. Control injections of acetone (i.e. the solvent for JH III) into the prothoracic ganglion do not significantly change the recipient females' phonotactic responses. Injection of JH III into the metathoracic ganglion also has no effect the females' phonotactic choices.     

Juvenile hormone and reproduction in crickets, Gryllus bimaculatus De Geer: corpus allatum activity (in vitro) in females during adult life cycle

ABSTRACT. Incubation conditions were established for a short-term radiochemical assay of spontaneous juvenile hormone (JH) biosynthesis in vitro by corpora allata from adult female Gryllus bimaculatus. The only JH synthesized was shown by HPLC to be JH III. A further incubation product, predominantly extracted from the corpora allata, was thought to be the JH III precursor, methyl farnesoate. In adult females reared at a constant temperature of 27°C the synthetic activities of the corpora allata-corpora cardiaca complexes in vitro increase from almost zero to a high peak value 4 days after the imaginal moult. Thereafter the activity decreases to varying intermediate levels, but always lower than the first maximum. Two days after the first peak in corpus allatum activity, ovarian fresh weight increases dramatically and the first oviposition occurs 2 days later.
Topical application of JH III to females reared at 20°C, which usually have a low fecundity, causes a dose-dependent stimulation of egg production and oviposition.     

Descending brain neurons in the cricket Gryllus bimaculatus (de Geer): auditory responses and impact on walking

The activity of four types of sound-sensitive descending brain neurons in the cricket Gryllus bimaculatus was recorded intracellularly while animals were standing or walking on an open-loop trackball system. In a neuron with a contralaterally descending axon, the male calling song elicited responses that copied the pulse pattern of the song during standing and walking. The accuracy of pulse copying increased during walking. Neurons with ipsilaterally descending axons responded weakly to sound only during standing. The responses were mainly to the first pulse of each chirp, whereas the complete pulse pattern of a chirp was not copied. During walking the auditory responses were suppressed in these neurons. The spiking activity of all four neuron types was significantly correlated to forward walking velocity, indicating their relevance for walking. Additionally, injection of depolarizing current elicited walking and/or steering in three of four neuron types described. In none of the neurons was the spiking activity both sufficient and necessary to elicit and maintain walking behaviour. Some neurons showed arborisations in the lateral accessory lobes, pointing to the relevance of this brain region for cricket audition and descending motor control.     

Expression of allatostatins in the Mediterranean field cricket, Gryllus bimaculatus de Geer (Ensifera, Gryllidae)

The allatostatin (AST) type A gene of the cricket Gryllus bimaculatus encodes a hormone precursor including at least 14 putative peptides with a common C-terminus Y/FXFGL/Iamide. By RT-PCR we have analyzed the expression of the allatostatin precursor in various tissues of 0-21 days old adult virgin and mated females. In 3-day-old virgin females, the gene is strongly expressed in the brain (oesophageal ganglion), the suboesophageal ganglion and the caecum, but to a lower extent in other parts of the digestive tract (ileum, midgut, colon), and in various other tissues such as the fat body, ovaries and female accessory reproductive glands. In the brain and ovaries of virgin females, the AST expression is rather constant throughout adult life, whereas in brains of mated animals, expression is low until day 7, but increases sharply from day 8 onwards to reach values triple those before day 7. In ovaries of mated animals AST gene expression is also age-dependent, with high expression rates during the first 4 days after imaginable moult, a second but smaller peak from day 15 to 21, and very low values in between. In the fat body of virgin crickets allatostatin expression is high during the first 9 days after ecdysis and declines thereafter, whereas in mated animals two peak values, day 1 and day 6, are observed, and a third peak in older animals.     

Diet affects female mating behaviour in a seed‐feeding beetle

In species where males provide nuptial gifts, females can improve their nutritional status and thus increase their fecundity by mating when in need of resources. However, mating can be costly, so females should only mate to acquire resources when the need for resources is large, such as when females are nutritionally‐deprived. Two populations of the seed‐feeding beetle Callosobruchus maculatus, a species that produces relatively large nuptial gifts, are used to test whether female nutritional status affects mating behaviour. Female access to water, sugar and yeast are manipulated and the fitness consequences of these manipulations are examined together with the effects of diet on the propensity of nonvirgin females to mate. Access to water has a small but significant effect on mass loss over time, lifespan and fecundity of females, relative to unfed controls. Access to sugar (dissolved in water) improves female fecundity and lifespan above that of hydrated females but access to yeast has no positive effects on female survival or reproduction. Diet has a large effect on both receptivity of nonvirgin females to a male and how quickly they accept that male. Unfed females are both more likely to mate, and accept a mate more quickly, than females provided access to water, which are more likely to mate and accept a mate more quickly than females provided with sugar. This rank order of behaviours matches the order predicted if females increase their mating rate when nutritionally deprived (i.e. it matches the effect of diet on female fitness). The results obtained also suggest that mate choice may be condition‐dependent: females from one population (Burkina Faso) show a preference for large males when well‐fed but not when unfed, although this result is not found in a second population (South India). It is concluded that nutritionally‐deprived females are more receptive to mates than are well‐fed females, consistent with the hypothesis that females ‘forage’ for nuptial gifts, or at least more willingly accept sperm in exchange for nuptial gifts, when they are nutritionally deprived.     

The influence of auditory and visual experience on the phonotactic behavior of the cricket,Gryllus bimaculatus

Female crickets lacking experience with phonotaxis to conspecific calling song respond to trains of continuously repeated sound pulses (trill), whereas experienced females do not. In the present study such inexperienced crickets were tested for their responsiveness to trills of pulse repetition periods from 30 to 70 ms on a Y- shaped maze. Stimulation with a repetition period of 30 ms led to unexpectedly low phonotactic and exploratory activity. Initial stimulation with trills of 30- ms repetition period drastically reduced the responsiveness of inexperienced animals to conspecific calling song and other attractive stimuli. Effects of visual stimulation on the phonotactic behavior of female crickets are demonstrated. Threatening visual stimuli changed the behavior of experienced animals to a state that resembles that of inexperienced animals. The relevance of these observations is discussed with respect to the development of the auditory pattern recognition mechanism in crickets.