Genetic evidence for multiple parentage in eastern kingbirds (Tyrannus tyrannus)

Summary Electrophoretic analysis was performed on 28 families of eastern kingbirds (Tyrannus tyrannus) from northern Michigan to estimate the occurrence of multiple parentage. Out of 19 families used in the final analysis, at least one putative parent was excluded in 9 families, or 18 out of 60 offspring (30% of offspring). Distribution of exclusion types conforms most closely to a model of quasi-parasitism, rather than extra-pair fertilizations, with secondary females laying their eggs in the primary female's nest, but random brood parasitism cannot be ruled out as an additional or alternative source of stray genes. Based on the model of random parasitism, an estimated 39% of all offspring in this population may be unrelated to one or both of the putative parents, or 53% based on a model of quasi-parasitism. Heretofore, eastern kingbirds have been considered to be exclusively monogamous; no behavioral evidence for alternative reproductive strategies has ever been reported for this species.     

Group-living in cliff swallows as an advantage in avoiding predators

Summary Cliff swallows (Hirundo pyrrhonota) in SW Nebraska, USA, nest in colonies and associate in groups away from their colonies. The degree to which group-living in this species affords advantages in the avoiding of predators was examined. The distance from the colony at which a snake predator was detected increased with colony size. In flocks away from the colonies, group vigilance increased, but the time that each individual spent vigilant decreased, with flock size. As a result, birds in large flocks had more time for preening and mud-gathering. Cliff swallows did not effectively mob predators and thus were unable to deter predators regardless of group size. Nesting within each colony was highly synchronous, but when the effects of ectoparasites on nesting success were removed, individuals nesting during the peak breeding period were no more successful than those nesting before or after the peak. This suggests that swamping of predators is unlikely in cliff swallow colonies. Nests at the edges of colonies were more likely to be preyed upon than nests nearer the center, suggesting that colonial nesting conferred some selfish herd benefits. Overall reproductive success did not vary with colony size. While cliff swallows receive some anti-predator benefits by living in groups, the avoidance of predators is probably not a major selective force for the evolution of coloniality in this species.     

Weather-mediated natural selection on arrival time in cliff swallows (Petrochelidon pyrrhonota)

An unusually long period of cold weather in May 1996 caused extensive mortality among insectivorous cliff swallows (Petrochelidon pyrrhonota) in the northern and central Great Plains. We analyzed how viability selection affected spring arrival time in a migratory Nebraska population by comparing capture histories of survivors with those of birds known to have died and by documenting how arrival time changed in the year following the selection event. Surviving birds had significantly later first-capture dates (an index of arrival time) in the years prior to selection than those that died; a significant selection differential suggested directional selection for birds that arrived later. Colony sites were occupied significantly later following the selection event, and the distribution of first-capture dates in the season after selection was significantly shifted toward later arrivals. Offspring of the survivors tended to arrive later than birds of the same age prior to the selection event. While major weather-caused mortality events of this magnitude are rare in the study area, spells of cold weather severe enough to cause limited mortality are frequent in April and early May. At least 25 probable mortality events of varying severity were identified in the last 50 years based on climatological data. Periodic weather-mediated selection against early arrival constrains the cliff swallow’s breeding season and may partly prevent directional selection for earlier nesting. Received: 19 October 1999 / Received in revised form: 15 January 2000 / Accepted: 24 January 2000     

Genetic evidence for mixed parentage in nests of the emu (Dromaius novaehollandiae)

Parentage in emus (Dromaius novaehollandiae) was examined by microsatellite analysis using four independent loci. Of 106 chicks sampled in one breeding season from 18 nests, 54 (51%) were not fathered by the nesting male, 12 (11%) were not from the observed mate of the sitting male, and 9 (8%) represented intra-specific brood parasitism, having no alleles in common with either nest parent. Some males (11%) fathered all chicks in their nests, but the majority showed high levels of cuckoldry. Those males commencing incubation earliest in the season tended to have the highest levels of paternity in their own nests. These results reveal a high frequency of extra-pair fertilisations and resultant cuckoldry in a predominantly socially monogamous bird and support recent reports which have described the emu mating system as a complexity of polyandrous, promiscuous and monogamous behaviour. Parentage assignment of chicks resulting from extra-pair fertilisations revealed an evenly scattered pattern of paternity that did not show any particular male dominance in reproductive success. These results lead to a reassessment of behavioural observations of emus, the consequences of parentage distribution, and theories about mating systems and sexual selection. The frequency of extra-pair copulations and intra-specific brood parasitism suggests patterns of descent that differ greatly from those implied by social monogamy. Received: 27 November 1999 / Received in revised form: 8 January 2000 / Accepted: 8 February 2000     

Genetic parentage in the indigo bunting: a study using DNA fingerprinting

Summary Parentage of nestlings in a North Carolina population of indigo buntings (Passerina cyanea) was analyzed using DNA fingerprinting. Three minisatellite DNA probes (wild type M13, Jeffreys' 33.15 and 33.6) were used to analyze nuclear DNA isolated from muscle biopsies of 63 nestlings, their parents, and other local adults. Each probe detected approximately 15 scorable fragments per individual, with 18%–39% overlap between probes. The proportion of bands shared (using all fragments over all three probes) between presumably unrelated adults averaged 0.23. Of the 63 offspring analyzed, 35 had at least one fragment not present in either putative parent. The distribution of offspring with novel fragments was distinctly bimodal. The lower mode (offspring with 0, 1, or 2 novel fragments, N=41) fit a Poisson distribution, a pattern expected if mutation (estimated rate per fragment= 0.01) were the source of the novel fragments. The remaining 22 offspring had more novel fragments than could be explained by mutation alone (minimum of four independent fragments across all three probes, =8.2). A low band-sharing proportion with the resident male ( =0.24) and high band-sharing with the resident female ( =0.60) implicated extra-pair fertilizations as the source of all 22. Thus in this sample, 35% of all nestlings came from extra-pair fertilizations and none from intra-specific brood parasitism. Of 25 broods sampled, 12 (48%) had at least one excluded offspring. In 3 broods all of the offspring excluded the resident male. Band-sharing proportions between excluded nestlings within a brood could not distinguish between single and multiple extra-pair paternity. Although young males tended to be excluded less often than older males, wing length and weight were not associated with the frequency of exclusion. Weight and wing length of females also were not associated with involvement in EPCs. Six of the 22 excluded offspring (in 3 broods) shared a large proportion of bands and had fewer than four novel fragments when compared to the fingerprints of a neighboring territorial male, implicating those males as actual fathers. The parentage of the remaining 16 offspring (in 9 broods) could not be clearly assigned because (1) one or more neighbors were not sampled or (2) difficulties in scoring across gels prevented confident alignment of fingerprint bands, and insufficient DNA was obtained from muscle samples to allow reanalysis of potential actual fathers on the same gel as excluded nestlings.     

Electrophoretic variants of egg white transferrin indicate a low rate of intraspecific brood parasitism in colonial cliff swallows in the Sierra Nevada,California

Summary The frequency of intraspecific brood parasitism in two colonies of cliff swallows (Hirundo pyrrhonota) nesting in the eastern Sierra Nevada in California was assessed through an electrophoretic analysis of egg white (albumin) proteins. Albumin proteins are maternally derived and are presumed to directly reflect maternal genotype. Thus, a comparison of protein banding patterns produced by eggs collected from a single clutch allows any egg laid by a female other than the putative mother to be detected. Eggs were collected from 13 cliff swallow nests in 1984 and 41 nests in 1987, a total of 54 nests. Of the clutches collected in 1984, one showed evidence of intraspecific brood parasitism, giving a nest parasitism rate of 7.6%. In 1987, one of 41 nests contained a parasitic egg, for a parasitism rate of 2.4%. Over both years combined the rate of intraspecific brood parasitism was 2 of 54 nests, or 3.7%. These parasitism rates are much lower than those estimated for Nebraskan cliff swallows (22–43%) and 2nd-year purple martins (36%). Possible explanations for the discrepancy between parasitism rates in Sierran cliff swallows and other groups of swallows are discussed.Correspondence to: A.P. Smyth     

Provisioning rules in tree swallows

Conflict between parents and offspring may result in offspring exaggerating their needs and parents devaluing their begging signals. To determine whether this occurs, it is first necessary to establish the link between need, begging and parental response. The purpose of our study was to examine these relationships in tree swallows (Tachycineta bicolor). Parents preferentially fed nestlings that begged sooner, reached higher and were closer to the front of the nestbox (Fig. 1). Begging intensity of both individuals and entire broods increased with relatively long periods between feeding visits. Within broods, parents responded to increased begging intensity by increasing their feeding rate, although this effect was relatively weak. Large and small nestlings did not differ in their begging behavior and all nestlings, regardless of size, were fed at similar rates. Despite the overall equity in feeding, male parents preferentially fed larger nestlings while female parents fed smaller nestlings. Nestlings did not increase their begging intensity in response to begging by nestmates. Our results suggest that begging is related to need in this species and that parents respond to variation in begging intensity. Received: 4 May 1995/Accepted after revision: 17 December 1995     

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     

Female control of extra-pair fertilization in tree swallows

Summary In a Canadian population of tree swallows, Tachycineta bicolor, DNA fingerprinting has previously shown that half of all broods contain many offspring resulting from extra-pair copulations (EPCs), whereas the other half contain only legitimate offspring. This bimodal pattern of extra-pair paternity might be due to variation in the effectiveness of male paternity guards, variation in female ability to resist EPCs, and/or variation in female pursuit of EPCs. Here we report experimental evidence for female control of copulations and fertilizations and the occurrence of two alternative copulation strategies among females in this population. Ten paired male tree swallows were removed on the day their mates laid the first egg. Replacement males took over the nestbox within 0.5–23 h and attempted to copulate with the widowed female. Assuming that eggs were fertilized approximately 24 h prior to laying, the first two eggs were fertilized before the male was removed, while the third and subsequent eggs could potentially be fertilized by the replacement male. Fingerprinting revealed that the first two eggs were sired by the resident males in five nests and by extra-pair males in the remaining five nests. The widows that had been faithful to their initially chosen mate rejected copulation attempts by the replacement male until most of the eggs had been laid. Consequently, nearly all eggs laid by these females were sired by the original male. The widows that had been unfaithful prior to male removal copulated sooner with the replacement male than females that were faithful to their mate. However, these replacement males also had a very low fertilization success; most eggs were sired by males that were not associated with the nest. This is consistent with the situation in non-experimental nests where unfaithful females copulate with their mate at the same rate as faithful females, yet unfaithful females have a majority of offspring sired by extra-pair males. We conclude that fertilization patterns to a large extent are determined by the female through active selection and rejection of copulation partners, though our results also allow some speculation that females have control over sperm competition. Female copulation tactics are probably determined some currently unknown fitness benefits of having the offspring sired by particular males.Correspondence to: Raleigh J. Robertson     

Genetic composition of a green turtle (Chelonia mydas) feeding ground population: evidence for multiple origins

Migratory marine turtles are extremely difficult to track between their feeding and nesting areas, and the link between juvenile and adult habitats is generally unknown. To assess the composition of a feeding ground (FG) population of juvenile green turtles (Cheloniamydas Linnaeus), mitochondrial DNA control region sequences were examined in 80 post-pelagic individuals (straight carapace length = 31 to 67 cm) sampled in September 1992 from Great Inagua, Bahamas, and compared to those of 194 individuals from nine Atlantic and Mediterranean nesting colonies. Evidence from genetic markers, haplotype frequencies, and maximum likelihood (ML) analyses are concordant in indicating that multiple colonies contribute to the Bahamian FG population. ML analyses suggested that most Bahamian FG juveniles originated in the western (79.5%) and eastern (12.9%) Caribbean regions, and these proportions are roughly comparable to the size of candidate rookeries. These data support a life-cycle model in which individuals become pooled in post-hatchling (pelagic) and juvenile (benthic) habitats as a consequence of ocean currents and movement among FGs. A substantial harvest of immature turtles on their feeding pastures will influence the reproductive success of contributing nesting populations over a wide geographic scale. Received: 1 April 1997 / Accepted: 14 October 1997     

Begging and provisioning in broods of asynchronously-hatched yellow-headed blackbird nestlings

Studies of begging have found a positive relationship between begging level and provisioning level. Studies of unequal nestlings, however, have found that small nestlings generally beg more but are fed less than their larger siblings. We manipulated the begging levels of yellow-headed blackbird (Xanthocephalus xanthocephalus) chicks to investigate how begging benefits individuals in broods of unequal siblings. Food-deprived chicks begged more and were fed more; satiated chicks begged less and were fed less. When we deprived each chick of a brood in turn, large and small chicks generally increased begging and received more provisioning. Small chicks, however, rarely received more food than their larger siblings even when they behged relatively more. Parent yellow-headed blackbirds increase provisioning to hungry begging chicks, but also allocate food based on relative offspring size.     

Need and nestmates affect begging in tree swallows

We conducted an experiment on nestling tree swallows (Tachycineta bicolor) to examine predictions from signalling models for the evolution of conspicuous begging behaviour. Specifically, we examined the relationship between (1) nestling begging intensity and hunger, (2) begging intensity and parental provisioning and (3) begging intensity and nestmate condition. Forty broods of 9-day-old nestlings were removed from their nests for 1 h and assigned to one of the following three treatments: (1) all nestlings in the brood deprived of food (n = 13), (2) all nestlings in the brood fed (n = 11) or (3) half the nestlings in the brood deprived and half fed (n = 16). Videotapes before and after the treatments showed that begging intensity increased in broods in which all of the nestlings had been deprived and decreased in broods in which all of the nestlings had been fed. Deprived nestlings in the half-and-half treatment did not change their begging intensity in response to treatment, while fed nestlings in this treatment group showed a decrease in begging intensity. Parent tree swallows increased their feeding rate to deprived broods and decreased their rate to fed broods. Within broods, parents decreased their feeding rate to fed nestlings, but showed no significant change in feeding to deprived nestlings. Our results suggest that begging intensity is influenced by hunger and that parents appear to respond to variation in begging intensity. The begging of nestmates also appears to influence begging independently of need. These results are consistent with predictions derived from signalling models of begging. Received: 20 June 1997 / Accepted after revision: 19 January 1998     

Patterns of male parentage in the fungus-growing ants

Ant queens from eight species, covering three genera of lower and two genera of higher attine ants, have exclusively or predominantly single mating. The ensuing full-sib colonies thus have a strong potential reproductive conflict between the queen and the workers over male production. This is because, all other things being equal, relatedness incentives should favour traits expressed in both workers and the queen to monopolise the production of the colony's male offspring. Microsatellite genotyping of males from these attine species shows that workers in queenless colonies are able to produce males, but that no worker-produced males were found in queenright colonies. Our results suggest that worker reproduction is rare or even absent in colonies with a fertile queen. This indicates that either the queen directly prevents the workers from raising their own sons, or that worker reproduction is absent in the presence of a fertile queen due to high ergonomic costs of this behaviour.     

Social hierarchy among siblings in broods of the oystercatcher Haematopus ostralegus

Summary On Skokholm Island, Wales, the young of the Oystercatcher (Haematopus ostralegus) are fully precocial, yet are totally dependent on parents for food. Though all young (commonly three) usually hatch within one day, small hatching intervals do exist, and first-hatched nestlings are usually slightly heavier at hatching than those hatched later. Firsthatched young and/or initially heavier young become socially dominant over their siblings; a stable, linear, non-aggressive social hierarchy among siblings develops, which determines the partition of food among them and results in unequal distribution when food is scarce. Food shortage impairs the growth rate of subordinate young. They are rather hungry and restless and consequently predation on them is higher than on dominant young.It is proposed that Oystercatchers achieve effective brood-reduction by the parents' control of differential egg weight, by the eggs' hatching order, and by the young's acceptance of a non-aggressive social hierarchy. The latter is an adaptation associated with the unusual phenomenon of fully precocial wader young being exclusively fed by parents.     

Multiple paternity and offspring quality in tree swallows

There is mounting evidence in a variety of taxa that females increase offspring quality by mating with multiple males, often resulting in multiple paternity. In birds, however, few studies have explicitly examined the benefits of mating with several different males; instead, the focus has been on whether or not extra-pair mating occurs, and its adaptive significance remains controversial. We examined the hypothesis that offspring quality, particularly immune response (phytohaemagglutinin assay) and growth, increases with the number of sires in broods of socially monogamous tree swallows (Tachycineta bicolor). We found one of the highest known levels of multiple paternity in birds (84% of nests with two or more extra-pair young had at least two extra-pair sires). Among nests with extra-pair young, the number and diversity of sires continued to increase linearly with the number of extra-pair young, so there was no evidence that some males monopolized paternity at high levels of extra-pair fertilization. Indeed, the number of sires was actually greater than expected in large broods, suggesting that some females might be seeking more mates. We found no effect of the number of sires on nestling immune response or growth. In mixed paternity broods, the immune response of extra-pair young did not differ from that of their within-pair half-siblings. However, among all broods, nestlings had a stronger immune response in nests with at least one extra-pair nestling than in nests with all within-pair nestlings. These results are not consistent with a good genes benefit of extra-pair mating, but they do suggest that there are environmental effects associated with extra-pair mating that increase nestling immune response. These environmental effects could produce indirect genetic effects on sexual selection if they are heritable. The extraordinarily high number of sires in this species highlights a relatively unexplored source of sexual selection in birds.     

Genetic evidence of self-fertilization in the sea anemone Epiactis prolifera

Biochemical genetic variation provided evidence for the mode of reproduction of brooded young in the sea anemone Epiactis prolifera Verrill, 1869. Individuals of E. prolifera are female when small but hermaphroditic when large (i.e., gynodioecious); juveniles are brooded externally on the column. Brooding individuals collected from 6 intertidal sites (5 in central California and 1 in Washington State, USA) in the spring and summer of 1980 were assayed for gene-enzyme variation by starch-gel electrophoresis. Three of 12 enzyme loci were polymorphic; phosphoglucose isomerase appeared to be encoded by two, closely linked loci. Genotypic frequencies deviated markedly from expected random mating proportions. Only three heterozygotes were found; two were heterozygous at all three polymorphic loci, and the other was polymorphic at the two PGI loci. All 158 juveniles from 25 brooding individuals were assayed (2–19 juveniles per parent). Juveniles on homozygous adults were always identical to their parent. However, brooded young of heterozygous individuals were not identical to their parent. but showed 1:2:1 phenotypic segregation ratios consistent with reproduction by self-fertilization. This genetic evidence together with findings of marked heterozygote deficiencies and genetic identity of homozygous adults and their brooded young supports the conclusion that E. prolifera usually reproduces by self-fertilization, and cross-fertilization is rate.     

Genetic evidence for monogamy in the cooperatively breeding red-cockaded woodpecker

We examined the genetic relationship among putative parents, offspring, and helpers in 224 red-cockaded woodpeckers (Picoides borealis) from the Sandhills of North Carolina. Comparison of DNA similarity with a pedigree constructed from 3,823 individually-marked birds observed from 1979 to 1992 provided verification of observed relatedness in the sampled population (R ² = 0.87,df = 14,P<0.001). In this population, breeding pairs may or may not have helpers, most of which are males that remain on their natal territory. Our sample included helpers assisting their parents, helpers assisting a related male and an unrelated female, and helpers assisting an unrelated pair. Band by band comparison and examination of similarity among DNA profiles indicated that all offspring from non-helped nests were sired by their putative parents (n = 28 families). Similarly, all but one offspring in helped nests were also sired by their putative parents (n = 16 families). In the exceptional case, the offspring evidently was sired by a male external to the group. Analysis of similarity values supported the conclusion that matings by helpers or extra-group males are rare or non-existent. Our results indicate that in this species advantages gained by individuals remaining on their natal territories as helpers do not generally include siring offspring. Correspondence to: S. Haig     

Genetic evidence for monogamy in the cooperatively breeding red-cockaded woodpecker

We examined the genetic relationship among putative parents, offspring, and helpers in 224 red-cock-aded woodpeckers (Picoides borealis) from the Sandhills of North Carolina. Comparison of DNA similarity with a pedigree constructed from 3,823 individually-marked birds observed from 1979 to 1992 provided verification of observed relatedness in the sampled population (R ² = 0.87, df = 14, P < 0.001). In this population, breeding pairs may or may not have helpers, most of which are males that remain on their natal territory. Our sample included helpers assisting their parents, helpers assisting a related male and an unrelated female, and helpers assisting an unrelated pair. Band by band comparison and examination of similarity among DNA profiles indicated that all offspring from non-helped nests were sired by their putative parents (n = 28 families). Similarly, all but one offspring in helped nests were also sired by their putative parents (n = 16 families). In the exceptional case, the offspring evidently was sired by a male external to the group. Analysis of similarity values supported the conclusion that matings by helpers or extra-group males are rare or non-existent. Our results indicate that in this species advantages gained by individuals remaining on their natal territories as helpers do not generally include siring offspring. Correspondence to: S. Haig