Sperm velocity in a promiscuous bird across experimental media of different viscosities.

In species with internal fertilization, the female genital tract appears challenging to sperm, possibly resulting from selection on for example ovarian fluid to control sperm behaviour and, ultimately, fertilization. Few studies, however, have examined the effects of swimming media viscosities on sperm performance. We quantified effects of media viscosities on sperm velocity in promiscuous willow warblers Phylloscopus trochilus. We used both a reaction norm and a character-state approach to model phenotypic plasticity of sperm behaviour across three experimental media of different viscosities. Compared with a standard medium (Dulbecco's Modified Eagle Medium, DMEM), media enriched with 1% or 2% w/v methyl cellulose decreased sperm velocity by up to about 50%. Spermatozoa from experimental ejaculates of different males responded similarly to different viscosities, and a lack of covariance between elevations and slopes of individual velocity-by-viscosity reaction norms indicated that spermatozoa from high- and low-velocity ejaculates were slowed down by a similar degree when confronted with high-viscosity environments. Positive cross-environment (1% versus 2% cellulose) covariances of sperm velocity under the character-state approach suggested that sperm performance represents a transitive trait, with rank order of individual ejaculates maintained when expressed against different environmental backgrounds. Importantly, however, a lack of significant covariances in sperm velocity involving a cellulose concentration of 0% indicated that pure DMEM represented a qualitatively different environment, questioning the validity of this widely used standard medium for assaying sperm performance. Enriching sperm environments along ecologically relevant gradients prior to assessing sperm performance will strengthen explanatory power of in vitro studies of sperm behaviour.

Sperm-related phenotypes implicated in both maintenance and breakdown of a natural species barrier in the house mouse.

The house mouse hybrid zone (HMHZ) is a species barrier thought to be maintained by a balance between dispersal and natural selection against hybrids. While the HMHZ is characterized by frequency discontinuities for some sex chromosome markers, there is an unexpected large-scale regional introgression of a Y chromosome across the barrier, in defiance of Haldane's rule. Recent work suggests that a major force maintaining the species barrier acts through sperm traits. Here, we test whether the Y chromosome penetration of the species barrier acts through sperm traits by assessing sperm characteristics of wild-caught males directly in a field laboratory set up in a Y introgression region of the HMHZ, later calculating the hybrid index of each male using 1401 diagnostic single nucleotide polymorphisms (SNPs). We found that both sperm count (SC) and sperm velocity were significantly reduced across the natural spectrum of hybrids. However, SC was more than rescued in the presence of the invading Y. Our results imply an asymmetric advantage for Y chromosome introgression consistent with the observed large-scale introgression. We suggest that selection on sperm-related traits probably explains a large component of patterns observed in the natural hybrid zone, including the Y chromosome penetration.

When Older Males Sire More Offspring-Increased Attractiveness or Higher Fertility?

Abstract: In birds with extrapair mating, older males usually have higher fertilization success than younger males. Two hypotheses can potentially explain this pattern: 1) females prefer older, and often more ornamented males, or 2) older males invest more in reproduction and fertility than younger males. Here we studied factors associated with age-related male fertilization success in a population of barn swallows Hirundo rustica in Canada. We document that male fertilization success increased gradually up to a minimum age of four-year old. The age effect was especially strong for the number of extrapair offspring obtained and the occurrence of a second brood. The higher fertilization success of older males was also associated with an early start of breeding in spring. The length of the elongated outermost tail feathers, a postulated male ornament preferred by females, also increased with age (in both sexes), but it was not a significant predictor of male fertilization success within age classes. Male fertility traits, especially testis size, but also sperm motility and sperm velocity, increased significantly across age groups. Our results suggest that the higher fertilization success by older males is due to their higher reproductive investments and that their longer tails are an adaptation to early arrival on the breeding grounds. Significance statement: The barn swallow is a socially monogamous passerine with extensive extrapair mating. We found that males become more successful in siring both withinpair and extrapair offspring as they become older. Their increased fertilization success was associated with a higher reproductive effort as indicated by larger testes, more motile sperm, and an earlier start of breeding in spring. The length of the outer tail feathers increased with age in both sexes, but long tails did not enhance male fertilization success among males of the same age. Long tails are probably an adaptation to rapid migration and earlier arrival on the breeding grounds. Our findings suggest that the commonly observed age-related increase in male fertilization success in passerine birds is better explained by life history theory than by sexual selection theory. Supplementary Information: The online version contains supplementary material available at 10.1007/s00265-022-03170-0.

The effects of radiation on sperm swimming behavior depend on plasma oxidative status in the barn swallow (Hirundo rustica).

Sperm are highly susceptible to reactive oxygen species (ROS) that can damage sperm DNA and structure, resulting in reduced fertilizing capacity. Exposure to radioactive contamination can also impair sperm swimming behavior and fertilizing ability, both through a reduction of sperm DNA integrity and via an increased generation of reactive oxygen species (ROS). However, the relationship between individual oxidative status and sperm swimming behavior has never been investigated in any wild population of animals exposed to radioactive contamination. We studied the motility of sperm collected from barn swallows, Hirundo rustica, breeding under different levels of radioactive contamination following the Chernobyl accident in 1986, in relation to individual oxidative status. We tested the hypothesis that the degree of impairment of sperm swimming behavior by radioactive contamination depended on plasma antioxidant capacity, the level of reactive oxygen metabolites (ROMs) and oxidative stress (sensu Costantini et al. 2006), a better oxidative status being associated with higher sperm motility. Sperm behavior parameters were subjected to principal component (PC) analysis, which extracted four PCs explaining 86% of the variance in sperm motility. PC2, representing sperm with high track velocity and ample lateral head displacement, was significantly predicted by the interaction between radiation level and either oxidative damage or oxidative stress. Contrary to our predictions, the highest values of PC2 were associated with relatively high radiation levels, particularly for high levels of either ROMs or oxidative stress. In addition, there was a tendency for values of PC3 (representing the percent of motile sperm) and PC4 (representing slow sperm with high beat cross frequency) to depend on the interaction between radiation level and total plasma antioxidant protection. Our results confirm the importance of oxidative status in determining the genetic and physiological outcome of exposure to radioactive contamination, complementing previous studies relating sperm abnormality to circulating levels of specific antioxidants. Our results also complement previous evidence that oxidative damage of sperm was negatively related to sperm motility, thus indicating a possible trade-off in quenching pro-oxidant compounds in the plasma and the seminal fluid.

Genetic and potential non-genetic benefits increase offspring fitness of polyandrous females in non-resource based mating system.

BACKGROUND: The adaptive significance of female polyandry is currently under considerable debate. In non-resource based mating systems, indirect, i.e. genetic benefits have been proposed to be responsible for the fitness gain from polyandry. We studied the benefits of polyandry in the Arctic charr (Salvelinus alpinus) using an experimental design in which the material investments by the sires and maternal environmental effects were controlled. RESULTS: Embryonic mortality showed a strong paternal genetic component, and it was lower in polyandrously fertilized offspring (sperm competition of two males) than in monandrous fertilizations. We also found that high sperm velocity was associated with low offspring mortality, but not with the size of the offspring or their yolk volume. Although no male effect was found on the size of the offspring yolk reserves, yolk volume was higher in offspring from polyandrous matings than offspring of the either of the two males when mated monandrously. CONCLUSIONS: In support of the "good sperm hypothesis, we found that sperm velocity was positively associated with offspring fitness. In addition, our results suggest that polyandrous females gain genetic advantage (higher offspring survival) from this behavior, but that some benefits of polyandry (larger yolk volume) may not be explained solely by the additive genetic effects. This suggests that sperm competition environment may intensify the selection on genetically superior sperm which in turn may produce offspring that have superior yolk reserves. However, as high sperm velocity was not associated with larger yolk volume, it is possible that also some other non-genetic effects may contribute to offspring fitness. The potential role of polyandrous mating in inbreeding avoidance is discussed.

Experimental evidence for paternal effects on offspring growth rate in Arctic charr (Salvelinus alpinus).

Sexual selection theory predicts that females should choose males that signal viability and quality. However, few studies have found fitness benefits among females mating with highly ornamented males. Here, we use Arctic charr (Salvelinus alpinus), a teleost fish with no parental care, to investigate whether females could gain fitness benefits by mating with highly ornamented and large-sized males. Carotenoid-based coloration signalled by males during spawning is believed to be an indicator of good genes for this species. Paternal effects on offspring size (body length and dry body mass) were examined experimentally by crossing eggs and sperm in vitro from 12 females and 24 males in a split-brood design and raising larvae to 30 days past hatching. We clearly demonstrated that there was a relationship between offspring size and paternal coloration. However, a negative interaction between paternal length and coloration was evident for offspring length, indicating that positive effects of paternal coloration were only present for smaller males. Thus, the red spawning coloration of the male Arctic charr seems to be an indicator of good genes, but the effect of paternal coloration on offspring length, an indicator of 'offspring quality', is size dependent.

No support for cryptic choice by ovarian fluid in an external fertilizer.

Whether the ovarian fluid (OF) represents a selective environment influencing cryptic female choice was tested using an external fertilizer experiencing intense sperm competition and large effects of OF on sperm swimming behavior-the Arctic charr (Salvelinus alpinus). We physically separated the OF from the eggs of reproductively active females and reintroduced either their own OF or fluid from another female to the eggs. The eggs were then fertilized in vitro in a replicated split-brood design with sperm from two males under synchronized sperm competition trials, while also measuring sperm velocity of the individual males in the individual OFs. We found large effects of males, but no effect of females (i.e., eggs) on paternity, determined from microsatellites. More important, we found no effect of OF treatments on the relative paternity of the two competing males in each pair. This experimental setup does not provide support for the hypothesis that OF plays an important role as medium for cryptic female choice in charr. Power analyses revealed that our sample size is large enough to detect medium-sized changes in relative paternity (medium-sized effect sizes), but not large enough to detect small changes in relative paternity. More studies are needed before a conclusion can be drawn about OF's potential influence on paternity under sperm competition-even in charr.

Rapid adjustments of sperm characteristics in relation to social status.

Sperm competition models predict that males typically mating in disfavoured roles should be selected to compensate for their disadvantage by investing more into sperm. We studied the effect of rapid changes in social status on ejaculate investments during experimental trials with an externally fertilizing teleost--the Arctic charr (Salvelinus alpinus). We document that males becoming dominant produce less sperm with lower velocity, but have higher sex steroid concentrations than subordinate males. These differences in sperm characteristics seem mainly to result from a decreased investment in sperm among fish that become dominant compared to pre-trial levels. Moreover, these adjustments of sperm production and sperm velocity seem not to be traded against sperm longevity. Our results support theoretical models of sperm competition, as males forced to mate in disfavoured roles seem to invest more into ejaculate quality than males in favoured roles. Additionally, we are the first to report that males, in a species with status-dependent shifts in reproductive tactics, have evolved rapid tactic specific adjustments of sperm production and sperm velocity corresponding to what could be predicted from their reproductive roles.

Quantitative characteristics of Atlantic halibut, Hippoglossus hippoglossus L., semen throughout the reproductive season.

The overall objective of the study was to investigate changes in quantitative parameters of Atlantic halibut (Hippoglossus hippoglossus L.) semen throughout the reproductive season in order to systematize the knowledge about biology of Atlantic halibut spermatozoa. Semen samples were collected from February to May from broodstock males kept under either a natural or 3-month advanced photoperiod regime. Spermatozoa concentration, semen pH and osmolality, as well as spermatozoa motility parameters were investigated. The use of catheterization of sperm was examined. Also, fertilization tests were performed. We found that spermatozoa concentration increases in a linear-like mode towards the end of the spawning season, which correlated with a decrease in a number of spermatozoa motility parameters, including actual percentage of motile spermatozoa (MOT), curvilinear velocity (VCL) and straight-line velocity (VSL) of spermatozoa. A breakpoint in MOT occurred when spermatozoa reached a concentration in the range of 17-20 x 10(9) spermatozoa/mL. The fertilization ability of sperm from males kept under natural photoperiod decreased in April. Survival of embryos at 80 degrees days produced by fertilizing eggs of single female with sperm from natural photoperiod males was 88, 76 and 41% on April 09 and 17, and May 01, respectively, whereas using sperm from 3-month delayed photoperiod males for fertilizing eggs from the same female on April 27 resulted in 80% of surviving embryos, not differing significantly from the data from April 09. Physical decomposition of spermatozoa was observed towards the end of the season and it was related to an increase in the whole semen osmolality. Catheterization of semen did not improve spermatozoa motility parameters, however, it reduced the variation in recorded values, especially in the case of pH, caused by contamination with feces or urine. Post-seasonal decrease in spermatozoa concentration was likely related to intensive ageing processes. Based on the present study and available data by other researchers, a model of changes of quantitative parameters in Atlantic halibut semen throughout the reproductive season is proposed.

Chilled storage of semen from Atlantic halibut, Hippoglossus hippoglossus L. I: optimizing the protocol.

Asynchrony in gamete production between females and males, and decrease in semen quality towards the end of reproductive season make chilled short-term storage of Atlantic halibut, Hippoglossus hippoglossus, semen a desirable method to apply for artificial propagation of this fish species. The goal of the present study was to determine the critical physiochemical factors that affect the success of chilled storage of halibut spermatozoa, and to develop a reliable, simple and efficient protocol for the storage. The presence and type of gaseous atmosphere, dilution and type of diluent, dilution ratio, and additional factors including spermatozoa sedimentation and replenishing the storage medium were tested in relation to spermatozoa motility parameters. Also, fertilization tests were performed with stored semen. Normoxia (air atmosphere) conditions were superior to both hyperoxia (pure oxygen) and no gaseous atmosphere for chilled storage. Dilution of semen with a diluent was superior to incubating undiluted semen. A dilution factor of between 6 and 10 times the original semen volume resulted in the longest viability of stored spermatozoa. Preventing spermatozoa sedimentation through daily swirling of the samples was superior to weekly swirling, however the effect was negligible for the first month of storage. Replenishing the storage medium showed no advantage to incubating in unchanged medium. Semen diluted in modified Hanks' balanced salt solution 1:5-1:9, supplemented with antibiotics, and kept at 0-1 degrees C in Ziploc bag filled with air retained its viability for exceptionally long time. A decrease in the percentage of motile spermatozoa was observed after 43 days of storage, and a decrease in curvilinear velocity occurred after 15 days. Samples remained motile for at least 79 days of storage and the fertilization ability was retained for at least 70 days of storage. The results demonstrate a high potential of application of chilled storage of semen into reproduction programs in Atlantic halibut aquaculture.

Chilled storage of semen from Atlantic halibut, Hippoglossus hippoglossus L. II: effect of spermiation advancement, catheterization of semen, and production-scale application.

A method for in vitro storage of Atlantic halibut Hippoglossus hippoglossus L. semen can facilitate seed production. The study aimed at determining the effect of male spermiation advancement on viability of chilled stored spermatozoa. The use of catheterization of semen from the sperm duct was examined. Also, large volumes of semen were stored under sub-optimal production-like conditions in order to determine the suitability of the method into practical use. Semen was collected from two broodstocks: natural photoperiod males, being at the first phase of the reproductive season and 3-month advanced photoperiod males, being at the end of the reproductive season. Semen samples were diluted 1:5 (v/v) with modified Hanks' Balanced Salt Solution supplemented with antibiotics, and stored in Ziploc bags filled with air. Sperm motility parameters, assessed by computer-assisted sperm analysis (CASA), were assessed weekly. Experimental and production-scale fertilization trials were performed. Sperm samples from natural photoperiod males showed significantly longer viability under in vitro storage conditions than sperm from advanced photoperiod males. In the natural photoperiod group, the decrease in spermatozoa motility, curvilinear velocity and straight-line velocity occurred on day 50, 14, and 28 of storage, respectively. Spermatozoa from one of five males were still motile on day 80 of storage, and fertilization rates and embryo survival rates obtained using semen stored for 70 days did not differ from control values and they were significantly higher than values obtained with the use of fresh semen of the same male, but being at the end of reproductive season. Catheterization of semen showed no advantage to stripping the semen without a catheter, even for samples stored undiluted for 1 day of collection, before dilution. Under sub-optimal conditions, spermatozoa stored in large volumes (10-100mL of diluted semen) without any special treatments except for weekly swirling, remained viable for more than 1 month. Production-scale fertilizations with samples stored for 5-21 days resulted in high survivals of embryos and hatchlings. Because of its simplicity and efficiency, the method shows a high potential for use in commercial Atlantic halibut farming. It has already been applied to a halibut breeding programme for the next reproductive season at our research station.

Effects of chronic dietary petroleum exposure on reproductive development in polar cod (Boreogadus saida).

Increasing human activities in the Arctic raise the risk of petroleum pollution, thus posing an elevated risk for Arctic organisms to be chronically exposed to petroleum compounds. The endocrine disrupting properties of some of these compounds (i.e. polycyclic aromatic hydrocarbons [PAHs]) present in crude oil may have negative effects on the long and energy intensive reproductive development of polar cod (Boreogadus saida), an Arctic keystone species. In the present study, selected reproductive parameters were examined in feral polar cod exposed to crude oil via a natural diet (0.11, 0.57 and 1.14µg crude oil/g fish/day [corresponding to low, medium and high treatments, respectively]) for 31 weeks prior to spawning. Fish maturing in the current reproductive period made up 92% of the experimental population while 5% were immature and 3% were identified as resting fish. Phase I metabolism of PAHs, indicated by ethoxyresorufin-O-deethylase (EROD) activity, showed a dose-dependent increase in high and medium crude oil treatments at week 6 and 22, respectively. Decreasing EROD activity and increasing PAH bile metabolite concentrations over the experimental period may be explained by reproductive maturity stage. Significant alterations in sperm motility were observed in crude oil exposed males compared to the controls. The investigated somatic indices (gonad and hepatic), germ cell development and plasma steroid levels (estradiol-17ß [females], testosterone [males and females] and 11-ketotestosterone [males]) were not significantly altered by chronic dietary exposure to crude oil. The environmentally realistic doses polar cod were chronically exposed to in this study were likely not high enough to induce adverse effects in this ecologically important fish species. This study elucidated many baseline aspects of polar cod reproductive physiology and emphasized the influence of maturation state on biomarkers of PAH biotransformation (EROD and PAH bile metabolites).

Aspermy, sperm quality and radiation in Chernobyl birds.

BACKGROUND: Following the Chernobyl nuclear power plant accident, large amounts of radionuclides were emitted and spread in the environment. Animals living in such contaminated areas are predicted to suffer fitness costs including reductions in the quality and quantity of gametes. METHODOLOGY/PRINCIPAL FINDINGS: We studied whether aspermy and sperm quality were affected by radioactive contamination by examining ejaculates from wild caught birds breeding in areas varying in background radiation level by more than three orders of magnitude around Chernobyl, Ukraine. The frequency of males with aspermy increased logarithmically with radiation level. While 18.4% of males from contaminated areas had no sperm that was only the case for 3.0% of males from uncontaminated control areas. Furthermore, there were negative relationships between sperm quality as reflected by reduced sperm velocity and motility, respectively, and radiation. CONCLUSIONS/SIGNIFICANCE: Our results suggest that radioactive contamination around Chernobyl affects sperm production and quality. We are the first to report an interspecific difference in sperm quality in relation to radioactive contamination.

210Po, 210Pb, 40K and 137Cs in edible wild berries and mushrooms and ingestion doses to man from high consumption rates of these wild foods.

This paper discusses activity concentrations of (210)Po, (210)Pb, (40)K and (137)Cs in edible wild berries and mushrooms collected from Øvre Dividalen national park, Northern Norway and derives committed effective ingestion doses to man based on high consumption rates of these wild foods. Edible wild berries and mushrooms accumulated similar levels of (210)Pb, but mushrooms accumulated higher levels of (210)Po and (40)K than berries. There appears to be a clear difference in the ability of Leccinum spp. of fungi to accumulate (210)Po and/or translocate (210)Po to mushrooms compared to Russula spp. of fungi. Activity concentrations of (137)Cs in edible wild berries and mushrooms from Øvre Dividalen national park reflected the lower levels of fallout of this radionuclide in Northern Norway compared to more central areas following the Chernobyl accident. For mushrooms, ingestion doses are dominated by (210)Po, while for berries, (40)K is typically the main contributor to dose. Based on high consumption rates, ingestion doses arising from the combination of (210)Po, (210)Pb and (40)K were up to 0.05 mSv/a for berries and 0.50 mSv/a for mushrooms. Consumption of such wild foods may result in a significant contribution to total annual doses when consumed in large quantities, particularly when selecting mushrooms species that accumulate high activity concentrations of (210)Po.

Elevated mortality among birds in Chernobyl as judged from skewed age and sex ratios.

BACKGROUND: Radiation has negative effects on survival of animals including humans, although the generality of this claim is poorly documented under low-dose field conditions. Because females may suffer disproportionately from the effects of radiation on survival due to differences in sex roles during reproduction, radiation-induced mortality may result in male-skewed adult sex ratios. METHODOLOGY/PRINCIPAL FINDING: We estimated the effects of low-dose radiation on adult survival rates in birds by determining age ratios of adults captured in mist nets during the breeding season in relation to background radiation levels around Chernobyl and in nearby uncontaminated control areas. Age ratios were skewed towards yearlings, especially in the most contaminated areas, implying that adult survival rates were reduced in contaminated areas, and that populations in such areas could only be maintained through immigration from nearby uncontaminated areas. Differential mortality in females resulted in a strongly male-skewed sex ratio in the most contaminated areas. In addition, males sang disproportionately commonly in the most contaminated areas where the sex ratio was male skewed presumably because males had difficulty finding and acquiring mates when females were rare. The results were not caused by permanent emigration by females from the most contaminated areas because none of the recaptured birds had changed breeding site, and the proportion of individuals with morphological abnormalities did not differ significantly between the sexes for areas with normal and higher levels of contamination. CONCLUSIONS/SIGNIFICANCE: These findings are consistent with the hypothesis that the adult survival rate of female birds is particularly susceptible to the effects of low-dose radiation, resulting in male skewed sex ratios at high levels of radiation. Such skewed age ratios towards yearlings in contaminated areas are consistent with the hypothesis that an area exceeding 30,000 km(2) in Chernobyl's surroundings constitutes an ecological trap that causes dramatic excess mortality.

No evidence for pre-copulatory sexual selection on sperm length in a passerine bird.

There is growing evidence that post-copulatory sexual selection, mediated by sperm competition, influences the evolution of sperm phenotypes. Evidence for pre-copulatory sexual selection effects on sperm traits, on the other hand, is rather scarce. A recent paper on the pied flycatcher, Ficedula hypoleuca, reported phenotypic associations between sperm length and two sexually selected male traits, i.e. plumage colour and arrival date, thus invoking pre-copulatory sexual selection for longer sperm. We were unable to replicate these associations with a larger data set from the same and two additional study populations; sperm length was not significantly related to either male plumage colour or arrival date. Furthermore, there was no significant difference in sperm length between populations despite marked differences in male plumage colour. We also found some evidence against the previously held assumption of longer sperm being qualitatively superior; longer sperm swam at the same speed as shorter sperm, but were less able to maintain speed over time. We argue that both empirical evidence and theoretical considerations suggest that the evolution of sperm morphology is not primarily associated with pre-copulatory sexual selection on male secondary sexual traits in this or other passerine bird species. The relatively large between-male variation in sperm length in this species is probably due to relaxed post-copulatory sexual selection.

Chernobyl birds have smaller brains.

BACKGROUND: Animals living in areas contaminated by radioactive material from Chernobyl suffer from increased oxidative stress and low levels of antioxidants. Therefore, normal development of the nervous system is jeopardized as reflected by high frequencies of developmental errors, reduced brain size and impaired cognitive abilities in humans. Alternatively, associations between psychological effects and radiation have been attributed to post-traumatic stress in humans. METHODOLOGY/PRINCIPAL FINDING: Here we used an extensive sample of 550 birds belonging to 48 species to test the prediction that even in the absence of post-traumatic stress, there is a negative association between relative brain size and level of background radiation. We found a negative association between brain size as reflected by external head volume and level of background radiation, independent of structural body size and body mass. The observed reduction in brain size in relation to background radiation amounted to 5% across the range of almost a factor 5,000 in radiation level. Species differed significantly in reduction in brain size with increasing background radiation, and brain size was the only morphological character that showed a negative relationship with radiation. Brain size was significantly smaller in yearlings than in older individuals. CONCLUSIONS/SIGNIFICANCE: Low dose radiation can have significant effects on normal brain development as reflected by brain size and therefore potentially cognitive ability. The fact that brain size was smaller in yearlings than in older individuals implies that there was significant directional selection on brain size with individuals with larger brains experiencing a viability advantage.

Sperm velocity and its relation to social status in Arctic charr (Salvelinus alpinus).

Sperm velocity has been shown to be a major determinant of fertilization success of external fertilizers. Yet, sperm velocity varies both within and between ejaculates and only a small number of fast sperm cells within an ejaculate are likely to have the potential of fertilizing the eggs. Having such fast cells should be of special importance during sperm competition, particularly for subordinate males that may release their sperm later or further away from eggs, than dominants. We examined ejaculates of dominant and subordinate male Arctic charr (Salvelinus alpinus), a species with sperm competition. Yet, rather than examining just average sperm velocity values, the aim was to examine whether the fastest fraction of sperm cell from dominant and subordinate males differed in velocity. While there was no difference in the average sperm velocity between dominant and subordinate males, analysis of the fastest swimming sperm cells show that subordinate males have significantly higher initial sperm velocity than dominant males within the 10, 5 and 1% fastest sperm cells. That is, the difference in sperm velocity between dominant and subordinate charr is most predominant among the fastest sperm cells. In sum, this study emphasizes the importance of studying the fastest sperm cells in the ejaculates, as status-dependent differences in sperm velocity may not be detected using average values.

Comparative evidence for the evolution of sperm swimming speed by sperm competition and female sperm storage duration in passerine birds.

Sperm swimming speed is an important determinant of male fertility and sperm competitiveness. Despite its fundamental biological importance, the underlying evolutionary processes affecting this male reproductive trait are poorly understood. Using a comparative approach in a phylogenetic framework, we tested the predictions that sperm swim faster with (1) increased risk of sperm competition, (2) shorter duration of female sperm storage, and (3) increased sperm length. We recorded sperm swimming speed in 42 North American and European free-living passerine bird species, representing 35 genera and 16 families. We found that sperm swimming speed was positively related to the frequency of extrapair paternity (a proxy for the risk of sperm competition) and negatively associated with clutch size (a proxy for the duration of female sperm storage). Sperm swimming speed was unrelated to sperm length, although sperm length also increased with the frequency of extrapair paternity. These results suggest that sperm swimming speed and sperm length are not closely associated traits and evolve independently in response to sperm competition in passerine birds. Our findings emphasize the significance of both sperm competition and female sperm storage duration as evolutionary forces driving sperm swimming speed.