A field test of the effects of familiarity and relatedness on social associations and reproduction in prairie voles

Inbreeding depression is a well-documented phenomenon. In animals, one means of avoiding the costs of inbreeding is through the recognition and avoidance of kin as mates. Prairie voles (Microtus ochrogaster) are short-lived, socially monogamous rodents that demonstrate inbreeding depression in the laboratory. Field data indicate that pair formation in nature is opportunistic but pairing among close relatives seems uncommon. We examined the role of relatedness and familiarity on prairie vole social associations and reproduction by placing adult voles into 0.1-ha enclosures with familiar siblings, unfamiliar siblings, and unrelated, unfamiliar conspecifics. Live-trapping data indicated that indices of social pair bonding were random with respect to relatedness and familiarity. Among females whose litters were sired by a single male, litters were significantly more likely to be sired by unfamiliar than familiar males, but the number of litters sired by males that were unrelated to their partner was not different from the number of litters sired by males that were related to their partner. Additionally, females that produced offspring with familiar siblings were significantly more likely to have litters with multiple paternity than females not producing offspring with familiar siblings. However, multiple paternity was not influenced by relatedness of sires. Finally, older individuals were more likely to produce offspring with each other than with younger individuals. Our findings suggest that prior association is a more important mechanism of inbreeding avoidance than phenotype matching for prairie voles mating under ecologically relevant conditions.     

Sociality,Bateman’s gradients,and the polygynandrous genetic mating system of round-tailed ground squirrels (<Emphasis Type="Italic">Xerospermophilus tereticaudus</Emphasis>)

Historically, most mammals have been classified as polygynous; although recent molecular evidence suggests that many mammals may be polygynandrous, particularly the ground-dwelling sciurids. We genotyped 351 round-tailed ground squirrels (Xerospermophilus tereticaudus) using seven microsatellite loci to determine paternity in 31 litters from 2004 to 2007. Polygyny was evident in all years except in 2007, when the population size was reduced. Multiple paternity occurred in the majority of litters (55%) with 2.5 ± 0.26 sires/litter (n = 31). Forty-nine percent of resident males (n = 114) sired offspring, and of males that sired offspring (n = 56) 27% sired young in multiple litters in a single breeding season. Litter size was positively correlated with the number of sires. Through an indirect analysis of paternity, we found 21 litters (68%) with an average relatedness of 0.5 or less. Males had a greater opportunity for sexual selection (I _s = 1.60) than females (I _s = 0.40); Bateman’s gradient was also greater in males (1.07 ± 0.04, n = 56) than females (0.82 ± 0.08, n = 31). The mating system in round-tailed ground squirrels defined through genetic analyses and Bateman’s gradients is polygynandrous compared to the previously suggested polygynous mating system as established by behavioral observations and fits within the predictions of the ground squirrel sociality models. Upon evaluating the predictions of the sociality models among sciurid species, we found a negative relationship between the level of sociality with litter size and the average percentage of multiple paternity within a litter. Thus, recent genetic information and reclassification of mating systems support the predictions of the ground-dwelling squirrel sociality models.     

Frequency of multiple paternity varies between two populations of brown smoothhound shark,Mustelus henlei

Multiple paternity was recently observed in a population of the brown smoothhound shark, Mustelus henlei, from Las Barrancas, Baja California Sur, Mexico, with litters demonstrating the greatest percentage of multiple paternity for any shark species (93 % of litters and an average number of sires = 2.3). To determine whether this frequency is consistent elsewhere in the species’ range, 4 polymorphic microsatellite loci were used to determine the frequency of multiple paternity in 18 litters of M. henlei from Santa Catalina Island, CA, sampled in 2004, 2008, and 2012. Multiple paternity varied among sampling years with 2004 demonstrating multiple sires for 40 % of sampled litters (n = 10) with an average of 1.4 sires per litter and 2008/2012 demonstrating a total lack of multiply sired litters (n = 8). Although multiple paternity was detected in this study, the range of frequencies observed is lower than that observed in the Mexican population. Based on these findings, investigators should take location into consideration when assessing the existence of multiple paternity in future studies of elasmobranch species.     

Sperm competition and ejaculate investment in red squirrels (<Emphasis Type="Italic">Tamiasciurus hudsonicus</Emphasis>)

Sperm competition is widespread in mammals and occurs when ejaculates from two or more males compete within the female’s reproductive tract to fertilize the ova. Enlarged testes are associated with sperm competition because they produce sperm, but the accessory glands produce fluids and proteins that are also important for fertilization success. Sperm morphology can also have consequences for fertilization success because of its influence on sperm motility. Red squirrels engage in multiple mating, and thus sperm competition is likely. Here, we assess levels of multiple paternity in a natural population of red squirrels, test the prediction that testis size is correlated with size of the accessory glands and sperm morphometry, and test the prediction that ejaculate investment is condition-dependent. Five of six litters (83%) showed evidence of multiple paternity, indicating that sperm competition is likely to have occurred. Testis size was correlated with the size of all three accessory glands (prostate, seminal vesicle, epididymides), and there was a generally positive relationship between the size of the accessory glands and sperm length. Sperm morphology showed significant variation in size and shape among individual male squirrels. There was no evidence of condition dependence of testis size or the size of the accessory glands, but sperm midpiece length was negatively related to body condition. Further work should include determining the fitness consequence of variation in sperm morphometry, testis size, and accessory gland size, and determining the effects of variation in ejaculate investment on sperm motility.     

Multiple paternity in a wild population of northern water snakes,Nerodia sipedon

Summary For most animal species, a single mating is sufficient to fertilize all of a female's offspring. As a result, females do not usually increase their reproductive success with successive matings. However, multiple paternity has been discovered in many animal taxa. We demonstrate that the majority of female water snakes in a wild population mate with more than one male for each litter. Field observations indicated that a highly skewed operational sex ratio (3.6:1 M : F) during the breeding season, while not necessary for multiple paternity to occur, created ample opportunity for females to mate multiply. Protein electrophoretic analysis showed that at least 12 of 14 litters from naturally mated females had more than one father. Since male snakes can not force copulations, multiple matings seem likely to be the result of female choice. Offprint requests to: F.E. Barry     

Experimental assessment of ecological and phenotypic factors affecting male mating success and polyandry in northern watersnakes, Nerodia sipedon

To resolve conflicting field observations regarding the action of sexual selection, we used breeding experiments and paternity analysis of the 927 resulting offspring to assess how male size, condition, tail length, genetic similarity to the female, and variation in operational sex ratio (OSR) affected male reproductive success and the incidence of polyandry in northern watersnakes (Nerodia sipedon). Only size affected male mating success. Large males were more successful, but only when male size varied substantially and competition among males was intense (i.e., male-biased OSR). The conditional nature of the size advantage may explain why studies of free-living watersnakes have produced inconsistent results regarding the relationship between male size and mating success. Size differences between males did not affect the proportion of offspring each male sired within multiply sired litters. We found positive size-assortative mating, but only when the OSR was female biased, suggesting that smaller males had improved access to females when competition among males was reduced, but that competition with larger males still restricted mating opportunities of small males to less preferred, smaller females. Most litters (58%) were multiply sired and larger females were more likely to produce multiply sired litters, similar to free-living watersnakes. There was no association between the incidence of multiple paternity and OSR, however, suggesting that polyandry is not simply a function of opportunity, with females passively waiting for males to court them.     

Genetic evidence of promiscuity in Peromyscus leucopus

Summary We collected pregnant female Peromyscus leucopus from natural populations during the summer of 1987 and 1988 and allowed these females to give birth to their field-conceived young in the laboratory. Blood samples were taken by suborbital puncture and phenotypes of five genetic loci (Esterase-1, trasferrin, hemoglobin, albumin and 6-phosphogluconate dehydrogenase) were studied using horizontal starch-gel electrophoresis to detect multiple paternity in single litters. Only esterase-1 was found to be highly polymorphic, with four alleles in samples of both years. One litter out of 29 in 1987 and 6 litters out of 32 in 1988 contained three different paternal alleles and served as direct evidence of multiple paternity in the field. The proportion of females engaging in multiple matings in natural populations of P. leucopus, assuming that all males were involved in every multiple mating, is 25%–100% (mean 68%). Because it is unlikely that all males are involved in every multiple mating, the actual proportion of females engaging in multiple matings should be greater. Offprint requests to: X. Xia at his Canadian address     

Sexual cannibalism facilitates genital damage in <Emphasis Type="Italic">Argiope lobata</Emphasis> (Araneae: Araneidae)

Sperm competition is a potent driving force in evolution leading to a remarkable variety of male adaptations that prevent or reduce fertilization by rivals. An extraordinary defensive strategy against sperm competition has evolved in a number of web spiders where males break off parts of their paired genitalia in order to obstruct the copulatory openings of females (mating plug). A recent comparative analysis on the family level reports that genital damage is most frequent in species with sexual cannibalism although, as yet, a functional association between sexual cannibalism and genital damage has not been found. Using the moderately sexually cannibalistic orb-web spider Argiope lobata, we show for the first time that males cannibalized during their first copulation damaged their pedipalps with significantly higher probability (74%) than males that escaped (15%). Of all males that damaged their genitalia, 44% were able to place a genital fragment inside the copulatory opening of the female, resulting in a relatively low total plugging rate of 14%. Successful obstruction of the female copulatory opening reduced the share of paternity of subsequent males (P ₂ = 0.06%), thus, indicating that genital damage may have evolved as a response to sperm competition in this species as well. However, the low incidence of successful plugging and the strong relationship between sexual cannibalism and genital damage suggest that apart from paternity protection, the nature of genital damage in A. lobata is further shaped by sexual conflict or cryptic female choice.     

Why do female mice mate with multiple males?

Females often show multi-male mating (MMM), but the adaptive functions are unclear. We tested whether female house mice (Mus musculus musculus) show MMM when they can choose their mates without male coercion. We released 32 females into separate enclosures where they could choose to mate with two neighboring males that were restricted to their own territories. We also tested whether females increase MMM when the available males appeared unable to exclude intruders from their territories. To manipulate territorial intrusion, we introduced scent-marked tiles from the neighboring males into males' territories, or we rearranged tiles within males' own territories as a control. Each female was tested in treatment and control conditions and we conducted paternity analyses on the 57 litters produced. We found that 46 % of litters were multiply sired, indicating that multiple paternity is common when females can choose their mates. Intrusion did not increase multiple paternity, though multiple paternity was significantly greater in the first trial when the males were virgins compared to the second trial. Since virgin male mice are highly infanticidal, this finding is consistent with the infanticide avoidance hypothesis. We also found that multiple paternity was higher when competing males showed small differences in their amount of scent marking, suggesting that females reduce MMM when they can detect differences in males' quality. Finally, multiple paternity was associated with increased litter size but only in the intrusion treatment, which suggests that the effect of multiple paternity on offspring number is dependent on male–male interactions.     

Extra-pair paternity and the evolution of testis size in a behaviorally monogamous tropical mammal,the large treeshrew (<Emphasis Type="Italic">Tupaia tana</Emphasis>)

Monogamy is rare in mammals (<5% spp.) but occurs in greater frequency among primates (15%) and their close relatives, the treeshrews (100%; Order: Scandentia). Two genetic studies of parentage in monogamous primates revealed high rates of extra-pair paternity (EPP), but to date parentage has not been studied in a treeshrew species. I analyzed the genetic parentage of 22 offspring from two populations of large treeshrews in Sabah, Malaysia (NE Borneo), using seven autosomal microsatellite loci and one mitochondrial DNA marker. Half of these offspring were sired by males that were not the presumed partner of the mother (50% EPP), and three litters exhibited evidence of multiple paternity. However, comparative analysis indicated that the high rate of EPP in Tupaia tana is not associated with intense sperm competition. Relative testis size of treeshrews was similar to testis size in 22 primate species with uni-male mating systems but smaller than 44 primates with multi-male mating systems. After factoring out the effects of body size and phylogeny, I also found that the evolution of multi-male mating systems was significantly associated with the evolution of larger testis size. Male–female pairs of T. tana occupy joint territories but forage and sleep alone (“dispersed pair-living”), and I argue that this form of behavioral monogamy renders mate guarding ineffective. The adaptive advantages of behavioral monogamy likely differ from the advantages driving EPP in large treeshrews. However, small testis size suggests that behavioral monogamy is not masking a dispersed multi-male mating system in this species.     

Mating patterns and reproductive success in the bushy-tailed woodrat (Neotoma cinerea), as revealed by DNA fingerprinting

The mating patterns and reproductive success of the bushy-tailed woodrat (Neotoma cinerea) were investigated over a 3-year period (1992–1994) using DNA fingerprinting. Paternity was determined by genetic analysis of 58 juveniles of known maternity from 35 litters. Analysis of DNA fingerprints revealed that all offspring within a litter were fathered by a single male; the statistical probability of detecting multiple males mating with a female was high, indicating that multiple paternity would have been detected had it occurred. However, individual males did not father more than one litter from a given female either within or between years. At least 75% of females and 57% of males successfully produced offspring each year. The finding that all littermates are first-order relatives may contribute to the high level of female cooperation in this species. Received: 28 May 1997 / Accepted after revision: 22 March 1998     

The effect of multiple mating on genetic relatedness in larval aggregations of the imported willow leaf beetle (Plagiodera versicolora,Coleoptera: Chrysomelidae)

Summary The effect of multiple mating on the average genetic relatedness within socially interacting sib groups was studied in the imported willow leaf beetle. Using an electrophoretically determined genetic marker, it was demonstrated that between 49 and 96% of wild caught females produce egg clutches expressing multiple sires. Following theoretical predictions of Wade (1982), this information was used to estimate the harmonic mean number of sires per family and then the average relatedness within families. Average relatedness was estimated to be 0.38. An alternate form of data analysis suggested by Pamilo and Crozier (1982) did not require any knowledge of paternity and resulted in a slightly higher estimate of relatedness, 0.44.     

Variance in male and female reproductive success in a harem-polygynous mammal,the black-tailed prairie dog (Sciuridae: Cynomys ludovicianus)

Summary Black-tailed prairie dogs are colonial rodents that live in contiguous social groups called coteries. A typical coterie contains one adult (2 years old) male, three or four adult females, and several yearlings and juveniles of both sexes. A large coterie sometimes contains two or more adult males. Using detailed behavioral observations on 164 females (of which 160 copulated) and data from four polymorphic loci from parents and offspring of 121 litters, we examined the black-tail mating system.Most females (101/164=62%) copulated with a single adult male, and only 3 of the 102 litters with 2 offspring (3%) showed unequivocal evidence of multiple paternity. Adult males usually copulated with several different adult females.In one-male coteries, females usually copulated exclusively with the resident adult male (RAM) (82/112=73%); this trend was confirmed by electrophoresis of blood smaples. In multimale coteries, each female frequently copulated with at least two different RAMs (28/52=54%); in 4 of 5 multimale coteries (80%) which produced two or more litters whose paternities could be unequivocally resolved by electrophoresis, two different RAMs each sired at least one litter.Of the 164 females, 30 (18%) copulated with both the RAM (or one of the RAMs, in multimale coteries) and an extracoterie adult male, but only 3 (2%) copulated exclusively with an extracoterie adult male. Electrophoresis showed that 9 of 121 litters (7%) were sired by an extracoterie adult male.Intersexual comparisons of annual reproductive success and lifetime reproductive success both indicate that black-tails are polygynous (i.e., that variance in reproductive success is greater for males than for females).     

Sex ratio determination and worker reproduction in the slave-making ant Harpagoxenus sublaevis

Summary In a population of the monogynous slave-making ant Harpagoxenus sublaevis in S.E. Sweden, the mean proportion of dry weight investment in queens was 0.54. This result differed significantly from 0.75 but not from 0.5, matching the prediction from the genetic relatedness hypothesis of sex ratio applied to slave-makers, given (as confirmed by this study) single mating of queens, population-wide mate competition, and relatively low levels of worker male production. Sex investment appeared unaffected by resource availability. In the same 47 colony population sample, fertile slave-maker workers were found in every queenless colony (ca. 30% of all colonies), and in 58% of queen-right colonies. Fertile workers occurred at a significantly higher frequency in the queenless colonies (19.2%) than in the queenright ones (9.8%), confirming that queenless conditions promote worker fertility. Fertile and sterile workers were similar in size. Electrophoretic allozyme analysis of ants from 49 colonies showed that: 1) queens mated singly; 2) female nestmates were full sisters (their regression coefficient of relatedness (±SE) was 0.735±0.044); 3) inbreeding did not occur; 4) queen and worker siblings were not genetically differentiated. Worker male production in queenright colonies was neither confirmed nor ruled out by the genetic data. However, production data indicated that queenless workers produced between 4.4 and 21.6% of all males. Overall colony productivity was largely determined by slave number, itself positively correlated with the number of slave-maker workers. There was an abrupt switch from all worker to all sexual production as colony size rose, as predicted by life history models. In queenright colonies, fertile slave-makers did not discernibly reduce colony productivity. Such workers occurred in queenright colonies with most slaves, suggesting they exploited energetic surpluses. Worker reproduction in H. sublaevis therefore appears to have greater influence at the level of individual behaviour than at colony or population level.     

Multiple paternity in meadow voles (Microtus pennsylvanicus): investigating the role of the female

Multiple paternity in single litters conceived in the wild was recently demonstrated in meadow voles (Microtuspennsylvanicus). In this study, we used an experimental approach (males tethered and females allowed to mate freely with one or several males) to investigate the role of female meadow voles in multiple paternity. We found that among 29 (of 39) females that copulated during our experiment, 79.3% chose to mate with more than one male. Female behavior in meadow voles thus clearly promotes multiple paternity and their role is an active one. Some of the hypotheses explaining promiscuity in meadow voles should be reconsidered in light of this result. We do not know the primary determinant of female mate choice, but male body mass played a secondary role in driving female preferences. The partial dependence between male body mass and female choice, coupled with the active role played by females, indicates that intersexual selection has the potential for reinforcing the effects of intrasexual selection (male-male dominance relationships) in this species. Finally, we demonstrate that the time period over which tests are conducted is an important part of the design of experiments aimed at understanding the role of females in multiple paternity. Received: 14 April 1998 / Accepted after revision: 12 September 1998     

Environmental,social, morphological,and behavioral constraints on opportunistic multiple paternity

Multiple mating and multiple paternity in polytocous species have been mostly studied from an adaptive (i.e., cost–benefit) perspective. Disease, time, energy, and the risk of injuries are well-known costs of multiple mating, yet from both male and female perspectives, a number of genetic and non-genetic benefits have also been identified. The effects of environmental conditions and individual-specific behavior, however, are much less well understood. Using a long-term study on yellow-bellied marmots (Marmota flaviventris), we evaluated the impacts of environmental variation, social structure, female body mass, and female docility (a personality trait) on the occurrence of multiple paternity. Multiple paternity was influenced by environmental constraints, social constraints, a female’s personality, and her body mass at emergence from hibernation. Personality and mass effects were detected only when environmental or social conditions were favorable. Our results suggest that multiple paternity is mainly limited by the opportunity to have access to multiple mates and is influenced by costs or mate choice because heavier females were more likely to have litters with multiple sires than smaller ones. Future studies in other species might benefit from considering environmental constraints when studying multiple paternity.     

Inbreeding and local mate competition in the ant Cardiocondyla batesii

The ant species Cardiocondyla batesii is unique in that, in contrast to all other ant species, both sexes are flightless. Female sexuals and wingless, ergatoid males mate in the nest in autumn and young queens disperse on foot to found their own colonies in spring. The close genetic relatedness between queens and their mates (r_qm=0.76±SE 0.12) and the high inbreeding coefficient (F=0.55; 95%CI 0.45–0.65) suggest that 83% of all matings are between brothers and sisters. As expected from local mate competition theory, sex ratios were extremely female biased, with more than 85% of all sexuals produced being young queens. Despite the common occurrence of inbreeding, we could not detect any adult diploid males. Though the probability of not detecting multiple mating was relatively high, at least one-third of all queens in our sample had mated more than once. Multiple mating to some extent counteracts the effects of inbreeding on worker relatedness (r_ww=0.68±SE 0.05) and would also be beneficial through decreasing diploid male load, if sex was determined by a single locus complementary system.Communicated by L. Sundström     

Low paternity in the hornet Vespa crabro indicates that multiple mating by queens is derived in vespine wasps

Queen mating frequency was studied in the European hornet, Vespa crabro, by analyzing four DNA microsatellite loci in 20 workers from each of 14 nests. Queens were found to be predominantly singly mated (9/14), although double (4/14) and triple mating (1/14) also occurred. For most multiply mated queens, paternity was significantly biased with the majority male fathering on average 80% of the female offspring. The population-wide effective mating frequency was therefore low (1.11), and sister-sister relatedness high (0.701 ± 0.023 SE). Low effective mating frequency in Vespa, in combination with data from other vespines, suggests that high paternity frequency is derived in the group. Some problems with the non-detection of fathers, where the queen was not sampled or shared alleles with males, are analyzed. Received: 16 November 1998 / Received in revised form: 29 March 1999 / Accepted: 12 April 1999     

Alpha-male paternity in elephant seals

The aim of this study was to assess paternity of males that dominated mating in harems at northern (Mirounga angustirostris) and southern (M. leonina) elephant seal rookeries using DNA fingerprinting and microsatellite DNA analysis. Southern alpha males had greater reproductive success than most northern alphas at similar-sized harems. Comparison of the relatedness between pups within harems also suggested that fewer males achieved matings in the southern elephant seal population. This was consistent with behavioral observations that suggest greater competition for mates in northern elephant seal harems. Reproductive success was consistent with estimates of mating success in some cases, but lower than expected for some northern elephant seal alpha males. A lower reproductive success than predicted from mating behavior may arise from a variety of factors including sperm competition, male sperm depletion from frequent mating, or reduced fertility. The alternatives are discussed in the context of environmental and historical factors. Received: 17 August 1998 / Received in revised form: 28 March 1999 / Accepted: 30 May 1999     

Mate replacement in experimentally widowed collared flycatchers (Ficedula albicollis): determinants and outcomes

There is increasing evidence that sexual selection may be intense even in socially monogamous birds, resulting from both mate choice and sperm competition. We studied these two modes of sexual selection experimentally by removing paired male collared flycatchers, Ficedula albicollis, from their mates for 2 days and investigating the factors that influenced the likelihood of a replacement male appearing and how the removals influenced paternity. Replacement males (usually neighbouring males) appeared at 81% (n = 37) of nests where males were removed. The likelihood of this appearance was unaffected by the probable reproductive value of the female's clutch to the replacing male. A replacement was, however, less likely when the original male had a large forehead patch, a trait previously shown to be subject to sexual selection in this population. Experimental removal of males increased the level of sperm competition: 74% of experimental broods were multiply sired, compared to 29% of unmanipulated broods in a previous study. Only two factors predicted how paternity was shared between males: removed males fathered more young if removed closer to laying, and if they had larger forehead patches. The former result is consistent with last-male sperm precedence determining paternity, whereas the latter adds to other evidence that forehead patch size is the target of female preference in this species. Our results suggest that females exert some control over male replacement, and also that they may influence the fertilisation success of males by behavioural means. Received: 15 July 1998 / Received in revised form: 16 March 1999 / Accepted: 28 March 1999