In the intimacy of the darkness: Genetic polyandry in deep-sea luminescent lanternsharks Etmopterus spinax and Etmopterus molleri (Squaliformes,Etmopteridae)

Multiple paternity seems common within elasmobranchs. Focusing on two deep-sea shark species, the velvet belly lanternshark (Etmopterus spinax) and the slendertail lanternshark (Etmopterus molleri) we inferred the paternity in 31 E. spinax litters from Norway (three to 18 embryos per litter) and six E. molleri litters from Japan (three to six embryos), using 21 and 10 specific microsatellites, respectively. At least two E. spinax litters were sired from multiple fathers each, with highly variable paternal skew (1:1 to 9:1). Conversely, no clear signal of genetic polyandry was found in E. molleri.     

Multiple paternity in a wild population of the yellow-necked mouseApodemus flavicollis

Multiple paternity has been described in a wide range of taxonomic groups (eg invertebrates, fish, reptiles, birds, mammals). In rodents, multiple paternity seems to be common and can lead to both genetic (eg increase in offspring diversity, avoiding inbreeding) and direct (eg higher survival rate of the litter) benefits. The primary aim of this study was to confirm multiple paternity and evaluate its frequency in a wild population of yellow-necked mouseApodemus flavicollis (Melchior, 1834). Animals were trapped in north-eastern Poland in 2004–2006. Five microsatellite loci previously described for members of the genusApodemus were used to examine the occurrence of multiple paternity among the offspring of 10 pregnant females. The analyses were performed using multiplex PCR, estimating the length of amplified fragments with an automated sequencer. The presence of additional alleles indicating multiple paternity was found in 30% (3 out of 10) of the investigated litters. Offspring fathered by a single male were predominant in each litter, with the proportion of individuals originating from other males varying from 16.7 to 20% in the three multiple paternity cases. Our findings indicate that the promiscuous mating system may be considered as an alternative breeding strategy in the yellow-necked mouse.     

Genetic analysis of the mating system and opportunity for sexual selection in northern water snakes (Nerodia sipedon)

We used data collected over 3 years at two study sites to quantifythe rates and consequences of multiple paternity and to determinethe opportunity for selection on male and female northern watersnakes (Nerodia sipedon). We sampled litters from 45 femalesthat gave birth to 811 offspring. Using eight microsatelliteDNA loci (probability of exclusion of nonparental males >0.99), we assigned paternity to 93% of neonates from one studypopulation and 69% of neonates from the other population. Observationsof participation in mating aggregations predicted individual reproductive success poorly for two reasons. First, males regularlycourted nonreproductive females. Second, more than half ofall sexually mature males obtained no reproductive successeach year, despite the fact that many of them participatedin mating aggregations. The number of sires per litter ranged from one to five, with 58% of all litters sired by more thanone male. Multiple paternity increased with female size, apparentlyboth because bigger females mated with more males and becausethe larger litters of big females provide paternity opportunitiesto more males. Multiple paternity was also more prevalent inyears with shorter mating seasons. We detected no advantage to multiple paternity in reducing either the number of unfertilizedovules or stillborn young. Despite the majority of males siringno young each year, some males fathered young with as manyas three different females in one year. Male reproductive successincreased by more than 10 offspring for each additional mate,whereas female success increased by fewer than 2 offspring foreach additional mate. The opportunity for sexual selectionwas more than five times higher in males than females.     

Multiple paternity in side-neck turtles Podocnemis expansa: evidence from microsatellite DNA data

Multiple paternity was found in two clutches of Podocnemis expansa using eight microsatellite loci. When loci were analysed separately a minimum of two males was estimated for nest N23, and three for nest C17. When all loci were combined, three patrilines were detected in N23, and six in C17. The distribution of full-sib cluster sizes indicated a disproportionate contribution of one male to clutch C17, consistent with possible sperm competition, or the mixing of leftover and newly acquired sperm. High mutation rates were detected at several loci. Multiple paternity has positive implications for this endangered species as it may slow the loss of genetic variability caused by drift. This is the first report of multiple paternity in the suborder Pleurodira.     

Paternity assessment in free-ranging wild boar (Sus scrofa) – Are littermates full-sibs?

Multiple paternity within litters occurs in various groups of mammals exhibiting different mating systems. Using seven genetic markers (i.e., microsatellites), we investigated the paternity of littermates in free-ranging wild boar (Sus scrofa) in a Mediterranean habitat. Using the software CERVUS 2.0 we estimated the probability of detecting multiple paternity across all loci (D), the probability of paternity (W) and a statistic Δ that allows the assignment of paternity to the most likely male with strict and relaxed levels of confidence. Multiple paternity was inferred for one of the nine analysed litters at the 80% confidence level. This suggests that a single male may control the access to receptive adult females and it shows that multiple paternity is not very common in the studied free-ranging wild boar population. Despite the possible occurrence of sperm competition and/or female cryptic choice, mate guarding seems to play a significant role in sexual selection. To better understand the wild boar's mating strategies further studies analysing the reproductive success of both sexes and under different environmental conditions should be conducted.     

Molecular phylogeny and distribution of three Apodemus species (Muridae, Rodentia) in Serbia

Phylogenetic relationships among three species of the genus Apodemus ( A. flavicollis, A. sylvaticus and A. agrarius ) from seven different localities in Serbia were estimated using arbitrarily primed PCR (AP-PCR) profiling, which allowed clear distinction between the sibling species A. flavicollis and A. sylvaticus . Eight oligonucleotide primers out of the 22 screened, gained a total of 397 amplified bands. The average pairwise genetic distances within Apodemus species ranged from 0.51 between A. flavicollis and A. sylvaticus , to 0.74 between A. sylvaticus and A. agrarius. The large genetic distances obtained among populations of A. flavicollis (0.13) indicate that this part of the Balkan Peninsula was probably a refuge during the last glaciation. Furthermore, differences obtained between populations south and north from the river Danube suggest that this species survived the Last Glacial Maximum in at least two separate refuges. Although our data for molecular markers are from a limited number of localities, extended analyses of C-band distribution on chromosomes from previously studied localities south of the rivers Danube and Sava, indicate that distribution of A. sylvaticus is limited to northern Serbia. Therefore, it is possible that this part of the Balkan Peninsula was not a refuge for this species during the last glaciation.     

High variation in multiple paternity of domestic cats (Felis catus L.) in relation to environmental conditions

Paternity was analysed in two domestic cat (Felis catus) populations differing in habitat structure (rural versus urban) and density (234 cats km-2 versus 2,091 cats km-2). A total of 312 offspring, 76 mothers and 65 putative fathers were typed at nine microsatellite loci in the two populations. Our data showed a high rate of multiple paternity in the urban population (70-83% of litters with more than one father), whereas it was much lower in the rural population (0-22% of litters with more than one), as most males were able to monopolize the entire litter. However, males reproduced as soon as they reached sexual maturity (i.e. ten months of age) in the urban population, whereas most males delayed reproduction to age three years in the rural population.     

Assessing multiple paternity in three commercially exploited shark species: Mustelus mustelus,Carcharhinus obscurus and Sphyrna lewini

In this study, multiple paternity (MP) was investigated in three commercially important shark species, common smoothhound Mustelus mustelus, dusky shark Carcharhinus obscurus and scalloped hammerhead Sphyrna lewini occurring in southern Africa. Reduced marker panels of between five and six microsatellite loci were constructed for each species and used to genotype and assess the presence of MP in a total of 60 M. mustelus individuals from six litters, 90 C. obscurus individuals from 14 litters and 54 S. lewini individuals from 13 litters. Analysis in GERUD and COLONY revealed the presence of MP in all three species. Multiple paternities were observed in 67, 35 and 46% of the litters of M. mustelus, C. obscurus and S. lewini, with corresponding average sire size of 1·6, 1·4 and 2·0, respectively. The variation in the rate of MP among the three species is in accordance with previous studies whilst the comparatively high frequency of MP observed for M. mustelus, matches what has previously been reported for shark species demonstrating aggregation behaviour.     

Multiple Paternity and Similar Variance in Reproductive Success of Male and Female Wood Mice (Apodemus sylvaticus) Housed in an Enclosure

The mating system and variance in individual reproductive success in wood mice ( Apodemus sylvaticus ) were analysed genetically and using observational studies within a large cage system in an outdoor enclosure. Four experimental groups contained four males and four females, each individually marked with a transponder (small computer chips injected under the skin) allowing individual detection of animals underground or within nest boxes without disturbance. The probability of paternity was analysed by comparing frequencies of cohabitation of males and females. In addition, DNA microsatellite analysis revealed reproductive success of each individual. Multiple paternity was found in 85% of all litters, which were sired by up to all four males. Males with a greater body mass, possibly indicative of a higher rank, sired more offspring than those with lower body mass. Interestingly, variance in the reproductive success of males and females did not differ. There was no indication that paternity could be assessed by the time males resided with a female shortly before she became pregnant. Our results indicate wood mice probably have a promiscuous mating system.     

A tale of two siblings: multiple paternity in big brown bats (Eptesicus fuscus) demonstrated using microsatellite markers

In many bat species, the opportunity for sperm competition or other mechanisms of post-copulatory paternity biasing is thought to be great, due to the long delay between copulation and fertilization, demonstrated sperm storage capabilities, and observed promiscuity. We present the results of the first study to assess whether litters of big brown bats (Eptesicus fuscus) containing dizygotic twins share the same father. We sampled 26 mother-offspring triads from three colonies in Indiana and Illinois, as well as 299 additional adults (237 females and 62 males) from these colonies and six other maternity colonies in the same area in 1997-1998. All individuals were genotyped at nine highly variable autosomal microsatellite loci and one X-linked locus. We assessed multiple paternity using autosomal and X-linked locus exclusions, and using maximum-likelihood methods. All methods confirmed multiple paternity within litters, and the maximum-likelihood analyses indicated that almost half of the sampled litters were composed of maternal half-siblings rather than full-siblings. Our results highlight the potential importance of post-copulatory mechanisms of paternity determination in the mating system of big brown bats, and have important implications for gene flow and population structuring in this species.     

Apodemus sylvaticus, a new host for Acanthocheilonema viteae (Nematoda: filarioidea)

Susceptibility of Apodemus sylvaticus and A. agrarius to infection with Acanthocheilonema viteae was compared with that of hamsters and jirds. Microfilaremia in A. sylvaticus was first noted on day 52 post-infection (p.i.) and lasted during the course of the study (up to day 150 p.i.). Maximum microfilaremic levels (female worm basis) of A. sylvaticus [mean +/- S.D. (n) = 690 +/- 1288(6)] were considerably higher than those of hamsters [16 +/- 18(6)] and jirds [51 +/- 25(5)]. Adult worm recovery in A. sylvaticus ranged from 2 to 40% of the number of infective larvae inoculated. Worm development in A. sylvaticus resembled that in hamsters and jirds. In contrast, microfilaremia was not detected in, nor adult worms recovered from A. agrarius throughout the study.     

Who's My Daddy? Considerations for the influence of sexual selection on multiple paternity in elasmobranch mating systems

Polyandry resulting in multiply‐sired litters has been documented in the majority of elasmobranch species examined to date. Although commonly observed, reasons for this mating system remain relatively obscure, especially in batoids. The round stingray (Urobatis halleri) is an abundant, well‐studied elasmobranch distributed throughout the northeastern Pacific that we used to explore hypotheses regarding multiple paternity in elasmobranchs. Twenty mid‐ to late‐term pregnant females were sampled off the coast of southern California and their litters analyzed for the occurrence of multiple paternity using five nuclear microsatellite loci. In addition, embryo sizes and their position within the female reproductive system (i.e., right or left uterus) were recorded and used to make inferences for patterns of ovulation. Multiple paternity was observed in 90% of litters and male reproductive success within litters was relatively even among sires. High variability in testes mass was observed suggesting that sperm competition is high in this species, although male reproductive success per litter appeared to be relatively even. Using embryo size as a proxy for fertilization, females were found to exhibit a variety of ovulation patterns that could function to limit a male's access to eggs and possibly promote high rates of multiple paternity. Our study highlights that elasmobranch mating systems may be more varied and complex than presumed and further investigation is warranted.     

Multiple paternity and mating patterns in the American alligator, Alligator mississippiensis

Eggs were sampled from 22 wild American alligator nests from the Rockefeller Wildlife Refuge in south-west Louisiana, along with the females guarding the nests. Three nests were sampled in 1995 and 19 were sampled in 1997. Females and offspring from all clutches were genotyped using five polymorphic microsatellite loci and the three nests from 1995 were also genotyped using one allozyme locus. Genotypes of the hatchlings were consistent with the guarding females being the mothers of their respective clutches. Multiple paternity was found in seven of the 22 clutches with one being fathered by three males, and the remaining six clutches having genotypes consistent with two males per clutch. Paternal contributions of multiply sired clutches were skewed. Some males sired hatchlings of more than one of the 22 clutches either as one of two sires of a multiple paternity clutch, as the sole sire of two different clutches, or as the sole sire of one clutch and one of two sires of a multiply sired clutch. There was no significant difference between females that had multiple paternity clutches and those that had singly sired clutches with respect to female total length (P = 0.844) and clutch size (P = 0.861). Also, there was no significant correlation between genetic relatedness of nesting females and pairwise nest distances (r2 = 0.003, F1,208 = 0.623, P = 0.431), indicating that females in this sample that nested close to one another were no more related than any two nesting females chosen at random. Eleven mutations were detected among hatchlings at the five loci over the 22 clutches. Most of these mutations (eight of 11) occurred at Ami(mu)-17, the only compound microsatellite locus of the five used in this study, corresponding to a mutation rate of 1.7 x 10-3. Finally, most of the mutations (82%) were homoplasious, i.e., mutating to an allelic state already present in this Louisiana population.     

Microsatellite markers and multiple paternity in the garter snake thamnophis sirtalis

Data from four microsatellite loci developed for the common garter snake, Thamnophis sirtalis, show that multiple paternity is common in a natural population on Beaver Island, Michigan. Six of eight litters tested, and all litters with five or more neonates, were multiply sired. At least triple paternity was documented in the largest litter examined (n = 13 neonates). Inheritance patterns and genotype frequencies in the wild population indicate the presence of null allele(s) at one of the microsatellite loci. Garter snakes are widely used in quantitative genetics research, and paternity testing is essential in studies that rely on sibling analysis.     

Multiple paternity in free-living root voles (Microtus oeconomus)

We used 10 microsatellite loci to determine the mating system and male reproductive success in a natural population of the root vole (Microtus oeconomus). By genotyping 21 females and their 111 offspring (5.28 ± 0.27 S.E. pups per female), we found evidence for multiple paternity in 38% of the litters sired by two or three males. Paternity was not significantly skewed away from the null expectation of equal proportions of offspring sired in any of the multiple-sired litters, and the most successful male fathered between 40% and 60% of the pups in a litter. The results indicate that promiscuity is a common mode of reproduction, consistent with the previous classification of the mating system based on the spatial structure of the root vole population.     

Geographic variation of multiple paternity in the American lobster, Homarus americanus

We studied the frequency of multiple paternity for American lobster (Homarus americanus) at three Canadian sites differing in exploitation rate and mean adult size. The probability of detecting multiple paternity using four microsatellite loci and 100 eggs per female was in excess of 99% under various scenarios of paternal contribution. Overall, 13% of the 108 examined females carried a clutch sired by two or three males. Multiple paternity was observed at the two most exploited sites (11% at Magdalen Islands and 28% at Grand Manan Island), whereas single paternity only was observed at the least exploited site (Anticosti Island). Within populations females with a clutch sired by more than one male tended to be smaller than females with a clutch sired by a single male. Based on these and other findings, we postulate a link between female promiscuity and sperm limitation in the American lobster.     

Multiple paternity in wild populations of invasive

Multiple paternity within litters has been recorded among a variety of small mammal species, including some species of rodents. Although multiple mating has been observed in wild Rattus populations, whether such mating results in litters with multiple paternities has not been established previously. For studies involving invasive species, awareness is useful of the level of genetic diversity a single pregnant invader can bring to a population. Multiple paternity is a means of providing additional genetic diversity to founding populations of rats on islands, which might improve population fitness. We used a genetic approach to confirm that multiple paternity occurs in wild populations of two rat species (Rattus norvegicus and R. rattus) in New?Zealand. This was accomplished by genotyping litters of embryos in pregnant females, and subtracting the known maternal alleles to find the number of paternal alleles necessary to form the litter. The number of paternal contributors cannot be overestimated by this method, befitting a conservative approach to the detection of multiple paternity, but can be underestimated. We used simulations to investigate the level of underestimation likely under two possible scenarios involving multiple paternity.     

Who's your daddy? Paternity testing reveals promiscuity and multiple paternity in the carnivorous marsupial Dasyurus maculatus (Marsupialia: Dasyuridae)

Female promiscuity is common among mammals but its advantages, particularly for marsupials, remain unclear. Using microsatellite DNA from pouch young of known mothers, we identified the most likely fathers of 25 wild spotted-tailed quolls ( Dasyurus maculatus ) from six litters. We aimed to determine whether young within the same litter had different fathers, and whether breeding success of males was associated with large body mass (consistent with inter-male competition) or scrotal width (consistent with sperm competition). We also explored the possible influence of promiscuity on relatedness within litters. Finally, we used data on paternity and relatedness to make inferences regarding movement and dispersal.
Four litters were sired by more than one male, and three males sired offspring in more than one litter. Known fathers had higher body mass, but not scrotal width, than males of unknown paternity status, suggesting that males may compete for access to females. Sires were less related to dams than expected by chance, and litters with multiple paternity had lower relatedness than litters sired by a single male.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 1–7.     

Genetic variation and differentiation in striped field mouse Apodemus agrarius inferred from RAPD-PCR analysis

Genetic variation and differentiation of the trans-Palearctic species Apodemus agrarius (striped field mouse), whose range consists of two large isolates-European-Siberian and Far Eastern-Chinese, were examined using RAPD-PCR analysis. The material from the both parts of the range was examined (41 individual of A. agrarius from 18 localities of Russia, Ukraine, Moldova, and Kazakhstan); the Far-Eastern part was represented by samples from the Amur region, Khabarovsk krai, and Primorye (Russia). Differences in frequencies of polymorphic RAPD loci were found between the European-Siberian and the Far Eastern population groups of striped field mouse. No "fixed" differences between them in RAPD spectra were found, and none of the used statistical methods permitted to distinguish with absolute certainty animals from the two range parts. Thus, genetic isolation of the European-Siberian and the Far Eastern population groups of A. agrarius is not strict. These results support the hypothesis on recent dispersal of striped field mouse from East to West Palearctics (during the Holocene climatic optimum, 7000 to 4500 years ago) and subsequent disjunction of the species range (not earlier than 4000-4500 years ago). The Far Eastern population group is more polymorphic than the European-Siberian one, while genetic heterogeneity is more uniformly distributed within it. This is probably explained by both historical events that happened during the species dispersal in the past, and different environmental conditions for the species in different parts of its range. The Far Eastern population group inhabits the area close to the distribution center of A. agrarius. It is likely that this group preserved genetic variation of the formerly integral ancestral form, while some amount of genetic polymorphism could be lost during the species colonization of the Siberian and European areas. To date, the settlement density and population number in general are higher than within the European-Siberian isolate, which seems to account for closer interpopulation associations, intense genetic exchange, and "smoothing" of polymorphism within the Far Eastern population group of A. agrarius.     

Multiple paternity in the European hedgehog

Previous observations have suggested that European hedgehogs Erinaceus europaeus are promiscuous, although there have been no direct studies of the mating system to confirm this. In this study, the genetic mating system of the European hedgehog was assessed by analysing mother–offspring groups using six polymorphic microsatellite loci. Evidence of multiple paternity was found in two of five litters analysed and therefore demonstrates the occurrence of polyandry in wild populations of hedgehogs.