A MICROSATELLITE ASSESSMENT OF SNEAKED FERTILIZATIONS AND EGG THIEVERY IN THE FIFTEENSPINE STICKLEBACK

Attempts by males to steal fertilizations from other males are common in many species. In some sticklebacks, males also are known to steal eggs from the nests of rivals and to carry them back to their own nests. However the genetic consequences of these nest-raiding behaviors seldom have been investigated. Here we assess genetically the prevalence of sneaked fertilizations and egg stealing, and we describe the mating system in a natural population of the fifteenspine stickleback. Six microsatellite markers were developed and employed to assay a total of 1307 embryos from 28 nests. Guardian males and all nest-holding males in the local area also were genotyped for two to six loci Analysis of male genotypes and those of embryos revealed that five of the 28 nests (18%) contained progeny from sneaked fertilizations, and that four of the 24 nests (17%) with resident males contained stolen egg clutches Comparisons of the composite DNA genotypes of nest-holding males against those of inferred sneakers implicated one nest holder as the sneaker of a nest seven meters from his own. Also, the genetic data demonstrated that nests of males frequently contain eggs from multiple females. The multilocus genotypes of inferred mothers indicated that females mate with multiple males, sometimes over distances greater than one kilometer.     

Social network predicts loss of fertilizations in nesting males of a fish with alternative reproductive tactics

Alternative reproductive tactics (ARTs) evolve when there is strong intra-sexual competition between conspecifics for access to mates. Typically, larger “bourgeois” males reproduce by securing the access to reproductive resources while smaller “parasitic” males reproduce by stealing fertilizations from larger males. A number of factors can influence the reproductive success of each tactic, including intrinsic (e.g. size) and extrinsic (e.g. tactic relative frequency) variables. An example where plastic ARTs occur is the peacock blenny Salaria pavo, with large males reproducing by defending nests and attracting females (bourgeois tactic) and small males reproducing by achieving sneaked fertilizations (parasitic tactic). In this study, we conducted field observations on individually tagged animals to determine their social network and collected eggs from 11 nests to determine the fertilization success of each male tactic. Paternity estimates for 550 offspring indicated an average fertilization success for nest-holder males of 95%. Nest-holder male morphological traits and social network parameters were tested as predictors of fertilization success, but only the number of sneakers present in the nest-holder’s social networks was found to be a predictor of paternity loss. Although male morphological traits had been previously found to be strongly correlated with reproductive success of nest-holder males, as measured by the number of eggs collected in the male’s nest, no correlation was found between any of the measured morphological traits and fertilization success for these males. The results suggest a stronger influence of the social environment than of morphological variables in the proportion of lost fertilizations by nest-holder males of this species.     

Bourgeois Males of the Peacock Blenny,Salaria pavo,Discriminate Female Mimics from Females?

In a Portuguese population of Salaria pavo, two types of reproductively active males occur: large bourgeois males that defend nests and have fully developed secondary sex characters (SSC) and small sneaker males that mimic the females’ morphology and behaviour to approach nests and parasitize fertilizations. These two alternative reproductive tactics are sequential, as sneakers develop into bourgeois males. We investigated whether bourgeois males were able to discriminate between sneakers and females of different sizes. In a laboratory experiment bourgeois males were sequentially presented with a female and a sneaker matched for size. Complementary field observations were conducted on the interactions between females and sneakers with bourgeois males. If bourgeois males discriminated females from sneakers, we predicted that they should direct more aggressive behaviours towards sneakers and more courtship acts towards females. In general, bourgeois males courted and attacked small females and sneakers equally. However, large sneakers were courted less and attacked more than large females. Larger sneakers, beginning to develop SSC, were attacked more and courted less by the bourgeois male than smaller sneakers, and larger females were attacked less and courted more than smaller females. These results strongly suggest that only small female‐mimicking males were able to deceive bourgeois males. In S. pavo, if the sneakers’ fertilization success correlates with their female‐mimicking ability, a fitness decrease with an increasing body size can be predicted.     

Extrapair paternity, inclusive fitness, and within-group benefits of helping in western bluebirds

In central coastal California, USA, 3–16% of western bluebird ( Sialia mexicana ) pairs have adult male helpers at the nest. Demographic data on a colour-ringed population over a 13-year period indicate that helpers gain a small indirect fitness benefit through increases in the number of young fledged from nests of close kin. A small proportion of adult helpers (16%) that were able to breed and help simultaneously had higher annual inclusive fitness than males that only bred. These males comprised such a minor proportion of helpers that the mean fitness of helpers was still lower than the mean fitness of independent breeders. We used DNA fingerprinting to determine whether extrapair fertilizations alter within-group benefits enough to tip the balance in favour of helping behaviour. Overall, 19% of 207 offspring were sired by males other than their social father and extrapair fertilizations occurred in 45% of 51 nests. Intraspecific brood parasitism was rare so that mean mother-nestling relatedness approximated the expected value of 0.5. Extrapair paternity reduced putative father-offspring relatedness to 0.38. Mean helper-nestling relatedness was 0.41 for helpers assisting one or both parents and 0.28 for helpers aiding their brothers. Helpers rarely sired offspring in the nests at which they helped. Helping was not conditional on paternity and helpers were not significantly more closely related to offspring in their parents' nests than to offspring in their own nests. Although helpers may derive extracurricular benefits if helping increases their own or their father's opportunities for extrapair fertilizations, within-nest inclusive fitness benefits of helping do not compensate males for failing to breed. Breeding failure and constraints on breeding are the most likely explanations for why most helpers help.     

MOLECULAR GENETIC DISSECTION OF SPAWNING,PARENTAGE, AND REPRODUCTIVE TACTICS IN A POPULATION OF REDBREAST SUNFISH,LEPOMIS AURITUS

Despite a great diversity of reproductive behaviors in fishes, few studies have examined the genetic consequences of alternative reproductive tactics. Here we develop and employ microsatellite markers to assess genetic paternity and maternity of progeny cohorts in a population of redbreast sunfish (Lepomis auritus), a species in which males build and tend nests. Nearly 1000 progeny from 25 nests, plus nest-attendant males and nearby adults, were genotyped at microsatellite loci that displayed more than 18 alleles each. The genetic data demonstrate that multiple females (at least two to six) spawned in each nest, their offspring were spatially dispersed across a nest, and more than 90% of the young were sired by the attendant male. However, about 40% of the nests also showed genetic evidence of low-level reproductive parasitism, and two nests were tended by males that had fathered none of the sampled offspring. Genetically deduced reproductive behaviors in this population of redbreast sunfish contrast with those reported previously in bluegill sunfish (L. macrochirus) wherein heteromorphic males specialized for parasitism or for parental care coexist in high frequency. Thus, nest-parasitic reproductive behaviors in fishes appear to be evolutionary labile.     

A molecular genetic examination of the mating system of pumpkinseed sunfish reveals high pay-offs for specialized sneakers

Intrasexual variation in reproductive behaviour and morphology are common in nature. Often, such variation appears to result from conditional strategies in which some individuals (e.g. younger males or those in poor condition) adopt a low pay-off phenotype as a 'best of a bad job'. Alternatively, reproductive polymorphisms can be maintained by balancing selection, with male phenotypes having equal fitnesses at equilibrium, but examples from nature are rare. Many species of sunfish (genus Lepomis) are thought to have alternative male reproductive behaviours, but most empirical work has focused on the bluegill sunfish and the mating systems of other sunfish remain poorly understood. We studied a population of pumpkinseed sunfish (Lepomis gibbosus) in upstate New York. Field observations confirm the existence of two male reproductive strategies: 'parentals' were relatively old and large males that maintained nests, and 'sneakers' were relatively young and small males that fertilize eggs by darting into nests of parentals during spawning. The sneaker and parental male strategies appear to be distinct life-history trajectories. Sneaker males represented 39% of the males observed spawning, and sneakers intruded on 43% of all mating attempts. Microsatellite analyses revealed that sneaker males fertilized an average of 15% of the eggs within a nest. This level of paternity by sneaker males appears to be higher than seen in most other fishes, and preliminary analyses suggest that the two male reproductive strategies are maintained as a balanced polymorphism.     

Coevolution of female fidelity and male help in populations with alternative reproductive tactics

In socially monogamous species, pair-bonded males often continue to provide care to all offspring in their nests despite some degree of paternity loss due to female extra-pair copulation. Previous theoretical models suggested that females can use their within-pair offspring as ‘hostages'' to blackmail their social mates, so that they continue to provide care to the brood at low levels of cuckoldry. These models, however, rely on the assumption of sufficiently accurate male detection of cuckoldry and the reduction of parental effort in case of suspicion. Therefore, they cannot explain the abundant cases where cuckolded males continue to provide extensive care to the brood. Here we use an analytical population genetics model and an individual-based simulation model to explore the coevolution of female fidelity and male help in populations with two genetically determined alternative reproductive tactics (ARTs): sneakers that achieve paternity solely via extra-pair copulations and bourgeois that form a mating pair and spend some efforts in brood care. We show that when the efficiency of mate guarding is intermediate, the bourgeois males can evolve to ‘specialize'' in providing care by spending more than 90% of time in helping their females while guarding them as much as possible, despite frequent cuckoldry by the sneakers. We also show that when sneakers have tactic-specific adaptations and thus are more competitive than the bourgeois in gaining extra-pair fertilizations, the frequency of sneakers and the degrees of female fidelity and male help can fluctuate in evolutionary cycles. Our theoretical predictions highlight the need for further empirical tests in species with ARTs.     

In Vitro Fertilization Reveals Offspring Recognition via Self-Referencing in a Fish with Paternal Care and Cuckoldry

Male bluegill sunfish (Lepomis macrochirus) exhibit alternative life histories: some males (parentals) delay maturation for up to 7 yr, then build nests, court females, and care for the eggs and fry, whereas other males (cuckolders) mature precociously, attempt to steal fertilizations from parentals, and provide no parental care. Parental males could avoid misdirecting their nepotism (i.e. caring for unrelated young) by abandoning entire broods if they were sired mainly by cuckolders or by discriminating between offspring and non‐kin fry within broods for which they care. We tested for kin discrimination by obtaining sperm from parental and cuckolder males and eggs from several females, and using them to conceive fry in vitro. In ‘blind’ laboratory tests, parental males (but not cuckolder males) distinguished between sources of dripping water that had been conditioned by their own offspring vs. unrelated fry. Parental males that were in the best physical condition were especially choosy. Because the only referents available to our experimental subjects were chemical cues emanating from their own body, our results imply that parental males can use self‐referent phenotype matching for kin recognition. This mechanism enables males to make the adaptive, nepotistic adjustments in paternal care that have been documented in previous studies.     

Effects of androgens on social behavior and morphology of alternative reproductive males of the Azorean rock-pool blenny

In the Azorean rock-pool blenny Parablennius sanguinolentus parvicornis two sequential reproductive tactics occur. Larger and older males establish breeding territories, while some of the smaller males become attached to nest-holder territories, acting as satellites on these territories, which they help to defend while trying parasitic fertilizations when females go in the nests to spawn. In the present paper we tested the effects of the androgens 17alpha-methyltestosterone (MT) and 11-ketotestosterone (KT) in the expression of male secondary sex characters and bourgeois behavior in satellite males. One week after satellites were implanted with Silastic tubes containing MT, KT, or castor oil (control), androgen-treated satellites had developed male secondary sex traits such as longer and wider male-type genital papilla and anal glands that secrete a sex pheromone, both traits being less expressed or absent, respectively, in satellite males. Androgen treatment had no effect on the gonadosomatic index or on the development of the testicular gland. KT treatment had a positive effect on relative liver weight. In terms of behavior, androgen-implanted individuals were less aggressive both in a mirror test and toward females when these were introduced into their tanks. MT-treated individuals spend more time inside the provided nests. Only androgen-implanted satellites managed to have the females entering their nests. When given a chance in a group tank either to try to attract females to their own nest or to act as satellites of an already established nest-holder's nest, MT-implanted males spent significantly more time in their own nest than near the nest-holder nest. These data suggest that androgens, particularly testosterone, may be involved in mating tactic switching in this species.     

Genetic evidence for cuckoldry in bluegill Lepomis macrochirus

In colonies of bluegill Lepomis macrochirus , some males provide parental care for broods in nests, whereas other males steal fertilizations and do not provide parental care. Using experimental pond populations of bluegill of known genotype (determined through protein electrophoresis), we demonstrate that cuckolder males successfully fertilize eggs in parental male nests. Using electrophoretic techniques to assess the fertilization success of nesting parental male bluegill in Lake Opinicon, Ontario, we demonstrate that paternity of these males ranges from 41% to 100% among four colonies studied. This difference among colonies is related to the density of cuckolder males.     

Extra-pair paternity in monogamous and polygynous Savannah sparrows,Passerculus sandwichensis

Extra-pair paternity can influence mating systems by affecting the fitness costs associated with polygyny. Polygyny is disadvantageous to males when the time and energetic demands of multiple pairings decrease either a male's success at gaining extra-pair fertilizations or his ability to ensure paternity among harem members. In Savannah sparrows,Passerculus sandwichensison Kent Island, New Brunswick, Canada, multilocus DNA fingerprinting of 136 adults and young revealed substantial female infidelity: overall, 31 of 92 young (33.7%) in 15 of 24 nests (62.5%) were the product of extra-pair fertilizations. Male mating status was a strong predictor of paternity. Each of seven monogamous females produced at least one extra-pair offspring, but only six of 11 primary females (54.5%) and two of six secondary females (33.3%) were unfaithful. As a result, nearly 80% of the young in nests of polygynous males resulted from within-pair fertilizations, compared with only 40% of the young in nests of monogamous males. Kent Island Savannah sparrows are simultaneously polygynous, and the absence of paternity costs associated with polygyny is surprising. The observed pattern of paternity suggests the operation of female choice, although male control of parentage cannot be excluded.     

Female choice and male competition in longear sunfish

Mating systems with spatial clumping of nests or display sitestheoretically can result from female preference for clumpedmales or from competitive interactions among males. We testedpredictions derived from these two types of models against observationsof nest dispersion, mating success, and alternative male matingtactics in a naturally occurring population of central longearsunfish (Lepomis megalotis megalotis). The data were consistentwith models for male-initiated spatial clumping of nest sites;no evidence of female preference for clumped nests was found.We observed that large males had greater mating success thansmall males and that solitarily nesting males were larger thancolonial males; the most successful males were solitary nesters.Mating success of large males did not increase with colony size.Furthermore, colonial males were often cuckolded by neighbors,whereas solitary males were not cuckolded. We propose that colonialnesting is maintained in longear sunfish because group nestingallows subordinate males to obtain access to females.     

The evolution of bourgeois, parasitic, and cooperative reproductive behaviors in fishes

Among vertebrate classes, fishes exhibit by far the greatest variability in competitive and cooperative behaviors in male reproduction. Scramble competition between reproductive males is one possibility. Another possibility occurs when resources, mates, or locations can be monopolized, in which case males may invest in primary access to fertilizations by adopting a "bourgeois" strategy, or they may employ alternative mating tactics to evade the reproductive monopoly of other males. Adaptations in morphology, physiology, and behavior to bourgeois and alternative phenotypes are highly divergent. Here I review the functional characteristics that differ between bourgeois and parasitic phenotypes, and discuss the variability of alternative reproductive tactics at the levels of plasticity, determination, and selection. Examples will illustrate the importance of ecology, and will suggest that variation in reproductive tactics is largely adaptive. Behavioral solutions to competition for mates and fertilizations often involve agonistic behavior and conflict, but also cooperation among competitors (e.g., when subordinate males pay a price to bourgeois males for gaining access to fertilizable eggs). Application of molecular genetic tools has helped to uncover intricate sexual and social relationships in various fish species, including species that display some of the most complex reproductive and social patterns known among the vertebrates.     

The effectiveness of mate guarding by male black-throated blue warblers

In many socially monogamous birds, males maintain close proximityto their mates during the fertile period. This is often consideredan effort on the male's part to prevent other males from copulatingwith his mate, but other functions have been suggested andthe effectiveness of males in preventing extrapair fertilizationshas come into question. Moreover, it is unclear whether mateguarding conflicts with other male activities, particularlythe pursuit of extrapair fertilizations. We examined mate guardingby male black-throated blue warblers (Dendroica caerulescens).Behavioral observations showed that males that guarded theirmates more closely were less likely to have extrapair youngin their nests. Moreover, the experimental detention of a malefor 1 h during the fertility risk period increased the probabilitythat a brood would contain extrapair young. Thus, male mate guarding was effective in reducing the risk of extrapair fertilization.Males with many opportunities for extrapair copulations appearedto guard their mates less and consequently had more extrapairyoung in their broods than males with few such opportunities.This suggests that mate guarding may conflict with the pursuitof extrapair fertilizations.     

The Relationship Between the Presence of Satellite Males and Nest-Holders' Mating Success in the Azorean Rock-Pool Blenny Parablennius sanguinolentus parvicornis

In the Azorean rock-pool blenny, sexually active males may adopt alternative reproductive tactics. In the present paper the relationship between the presence of satellite males and the reproductive success of nest-holders was investigated by comparing nests with and without an associated satellite male. Males with an associated satellite male suffered more conspecific intrusions but they did not display a higher frequency of attacks towards conspecifics. Nest-holder males were more aggressive towards other conspecifics than towards satellites and the tolerance of nest-holders towards satellites was inversely correlated with the time spent by the satellites in the breeding territory, which suggests control by the nest-holder male of the satellite investment in shared territorial defence. Nest-holders with an associated satellite male had higher condition factors and received more female visits and more spawnings. These results bear two possible interpretations. (1) Nest-holders benefit from the presence of a satellite male by increased attractiveness of their nests to females; satellite males are mutualists helping to defend the nest-owner's territory and to attract females, which is why they are tolerated. (2) Satellite males associate preferentially with more successful nest-holder males which have higher condition factors, and by doing so have more opportunities to achieve parasitic fertilizations. Only experiments will allow these two hypotheses to be distinguished.     

Photoperiodic determination of gynoparae and males of damson-hop aphid Phorodon humuli

Abstract. Sexual morph production in Phorodon humuli is controlled by daylength. Wingless aphids reared from birth in short-day conditions (LD 12 : 12 h) and transferred when adult to long-day conditions (LD 18 : 6 h) produce only gynoparae and males until death some 7 weeks later, whereas those reared in long days and transferred to short days produce 20% wingless parthenogenetic females, 50% gynoparae and 30% of males in an overlapping sequence. No winged morph capable of re-infesting hop is produced. Less mature embryos are more sensitive to short days than older embryos because 100% of the former became gynoparae after 4 days of exposure of their mothers, and 59% when their mothers were exposed for the 4 days immediately preceding the birth of their offspring. Two days of exposure to short days switches 94% of young embryos from wingless to gynopara production when mature. The response to short days is irreversible. Wingless aphids reared from birth to adult in short days produce 30% fewer offspring than those reared to the same stage in long days and are male-biased, with 76–78% of their offspring being male.     

Male sexual polymorphism, alternative reproductive tactics, and androgens in combtooth blennies (pisces: blenniidae)

In species in which intense intermale competition for the access to females is present males of lower competitive ability may adopt alternative reproductive tactics (ART) to get access to mates. These ART translate in many cases into male sexual polymorphism, with individuals following distinctly different tactics. Usually two alternative male morphs can be recognized in species with ART: (1) bourgeois males that compete for access to mates invest in typically male behaviors, such as building elaborated nests or displaying ornaments; and (2) parasitic males that take advantage of the success of the bourgeois males in attracting females and attempt "sneaker" fertilizations (e.g., sneaker and satellite males). In combtooth blennies (Blenniidae) the co-occurrence of ART and male sexual polymorphism has been described for two temperate species: the peacock blenny, Salaria pavo, and the Azorean rock-pool blenny, Parablennius sanguinolentus parvicornis. Interestingly, while in the peacock blenny the alternative male morph adopts a sneaker tactic, in the rock-pool blenny parasitic males act as satellites to nest-holder males. Thus, this variation in the ART expressed in these two closely related species allows for a comparative study of the proximate and ultimate factors affecting the expression of the two ART. In this article we summarize the available information on androgen levels in bourgeois and parasitic males of natural populations of the two species and of recent studies on the effect of exogenous administration of androgens on tactic switching in parasitic males of the two species. The information is discussed within the frame of the relative plasticity hypothesis, which predicts that plastic alternative morphs should show differences in hormone levels and that the administration of sex steroids should be effective in promoting the switch from the parasitic to bourgeois tactic. The evidence is only partly consistent with this hypothesis. Alternatively, a social transduction hypothesis that better fits the available data on androgens and ART in teleost is proposed. It states that the observed differences in androgen levels between alternative morphs should not be interpreted as an organization vs activation effect of steroids, but rather as the limited vs lifelong responsiveness of the neuroendocrine axis to social regulation.     

Maintenance of female mimicry as a reproductive strategy in bluegill sunfish (Lepomis macrochirus)

Synopsis In species where male reproductive success is dependent on male competition and aggression, alternative reproductive patterns, thought to represent a reduction in male reproductive effort, sometimes occur. Female mimicry in bluegill sunfish (Lepomis macrochirus) is an example of an obligate alternative male strategy. Female mimics are small, sexually mature males which mimic the details of female behavior, and gain access to functional females attracted to the nests of large, aggressive territorial males. The costs of female mimicry relative to nesting male behavior are discussed and two hypotheses, deception and mutual gain, are presented to explain the tolerance and courtship of female mimics by nesting males.     

Male share of provisioning is not influenced by actual or apparent loss of paternity in western bluebirds

Approximately 45% of western bluebird (Sialia mexicana) femaleshave some chicks in the nest that are not sired by their socialmates. Extrapair fertilizations account for 42% of offspringin these nests and 19% of nestlings overall. I tested the hypothesisthat males reduce nestling provisioning when their certaintyof paternity or share of paternity is reduced. Capture and detentionof socially monogamous males for 1 h or 24 h during the layingperiod reduced males' copulatory access and their ability tomate guard, increasing the frequency with which extrapair malesintruded and attempted to copulate with resident females. Malesdetained during laying did not reduce their share of feedingtrips compared to control males detained during incubation,compared to unmanipulated males, or compared to males that werecaptured but not detained. Males detained on territory for 1h during the laying period did not reduce their share of feedingtrips when they observed male intrusion, nor when they observedtheir mates accepting extrapair copulations. Males that witnessedtheir mates accepting extrapair copulations did not reduce theirshare of risk in provisioning. Genetic fingerprinting at nonexperimentalnests indicated that males also failed to reduce their feedingcontributions when their estimated share of paternity was reduced,even when a helper male was present to reduce the impact onnestlings. These results suggest that male western bluebirdsdo not make significant adjustments in their share of provisioningwhen they have evidence of partial paternity loss. Togetherwith prior results, this study suggests that western bluebirdmales use an all-or-none rule, contributing approximately halfof the parental provisioning at nests, as long they have somecopulatory access to the female during egg laying.     

Sex-specific embryonic mortality during concurrent pregnancy and lactation in house mice

To evaluate the importance of lactational stress for sex ratio manipulation in postpartum inseminated house mice, 163 sexually inexperienced male and female BALB/c ABom inbred mice were mated and stud males were removed the day following the initial parturition. Randomly chosen dams were allowed to suckle their young for either one or 14 or 21 days. The results showed that two weeks of lactation enhance the incidence of total abortion of the litter conceived postpartum. If lactation continued another week, dams which gave birth to a second litter lost relatively more embryos during uterine development and the percentage of lost embryos was increased by the number of pups suckled. Among these dams, the number of embryos lost in utero correlated positively with viable litter sex ratios (percentage male pups) at second parturition, indicating that lactational stress enhances embryonic mortality and that this additional mortality predominantly affects female embryos.