Patrick Pomeroy

Male mating success and paternity in the grey seal, Halichoerus grypus: a study using DNA fingerprinting

Grey seals breed colonially on remote coastal sites. Within the colony, males compete aggressively for access to the females. We compare field observations of breeding behaviour with paternity, as determined by DNA fingerprinting, in the breeding colony on the island of North Rona. In 89% of cases where paternity was assigned, the father was observed near the mother during her perioestrous period, although in some cases this was discovered retrospectively. However, the most likely candidate male, judged on the basis of behavioural criteria, was shown not to be the father in 36% of cases. Overall, DNA typed males were more dominant, maintained positions amongst the females for longer, and accounted for disproportionately more paternities than untyped males. However, the reproductive success of the typed males is not as great as their behavioural domination of copulatory opportunities would suggest. Possible contributory factors which could explain this include: (i) imprecision in the estimates of copulatory opportunity due to mobility of males or topographical influences on individual behaviour; and (ii) opportunities for subordinate males to copulate with receptive females, either sneakily within the colony or in the water.

The impact of climatic variation on the opportunity for sexual selection.

Many studies have demonstrated influences of climatic variation on a variety of ecological processes, however, its impact on the potent evolutionary force of sexual selection has largely been ignored. The intensity of sexual selection is a fundamental parameter in animal populations, which depends upon the degree of polygamy and will probably be influenced by the impact of local climatic variation upon ‘environmental potential for polygamy’. Here, we provide evidence of a direct effect of local climatic variation on the intensity of sexual selection, by showing a clear correlation between local weather conditions and inter-annual changes in the degree of polygamy in a long-term study of colonially breeding grey seals (Halichoerus grypus). Our results show that changes in local weather conditions alter the annual proportion of males contributing to the effective population size (Ne) by up to 61%. Consequently, over the ‘lifetime’ of a cohort, a broader range of individuals will contribute genetically to the next generation if local weather conditions are variable. In the context of predicted future changes in climatic variation, these findings have broad implications for population genetics of socially structured animal systems through the major influence that the degree of polygamy has upon Ne.

Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure

BACKGROUND The neuropeptide oxytocin is increasingly the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHOD This study determined the sample collection and analysis protocols that cause the least amounts of protocol dependant variation in plasma oxytocin concentrations detected by ELISA. The effect of vacutainer type, sample extraction prior to analysis and capture and restraint protocol were investigated while validating an assay protocol for two novel species, grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina). RESULTS Where samples are extracted prior to analysis, vacutainer type (EDTA mean: 8.25±0.56 pg/ml, heparin mean: 8.25±0.62 pg/ml, p=0.82), time taken to obtain a sample and restraint protocol did not affect the concentration of oxytocin detected. However, concentrations of oxytocin detected in raw plasma samples were significantly higher than those in extracted samples, and varied significantly with vacutainer type (EDTA mean: 534.4±43.7 pg/ml, heparin mean: 300.9±19.6 pg/ml, p<0.001) and capture and restraint methodology. There was no relationship between oxytocin concentrations detected in raw and extracted plasma (p=0.25). COMPARISON WITH EXISTING METHOD(S) Over half the reviewed published studies analysing plasma oxytocin use raw plasma and different vacutainer types are used without consistency or justification through-out the literature. CONCLUSIONS We caution that studies using raw plasma are likely to over estimate oxytocin concentrations, cannot be used to accurately infer true values via correlations and are susceptible to variation according vacutainer type.

Reproductive cycles of marine mammals.

Marine mammals conform to the general mammalian reproductive system centered on the hypothalamic-pituitary-gonadal axis. Most marine mammals are long-lived and of large body size with lesser reproductive rates than many other animals, a consequence of their interaction with the marine environment where the demands of acquiring resources from the ocean must be balanced with the need for bearing offspring in a suitable place for survival. The degree of spatial and temporal separation of these life history phases in many species is a key feature of their ecology. The reproductive physiology of pinnipeds, cetaceans, sirenians, sea otters and polar bears has been more thoroughly characterized for the more accessible species.

Estimating demographic parameters for capture–recapture data in the presence of multiple mark types

In mark-recapture studies, various techniques can be used to uniquely identify individual animals, such as ringing, tagging or photo-identification using natural markings. In some long-term studies more than one type of marking procedure may be implemented during the study period. In these circumstances, ignoring the different mark types can produce biased survival estimates since the assumption that the different mark types are equally catchable (homogeneous capture probability across mark types) may be incorrect. We implement an integrated approach where we simultaneously analyse data obtained using three different marking techniques, assuming that animals can be cross-classified across the different mark types. We discriminate between competing models using the AIC statistic. This technique also allows us to estimate both relative mark-loss probabilities and relative recapture efficiency rates for the different marking methods. We initially perform a simulation study to explore the different biases that can be introduced if we assume a homogeneous recapture probability over mark type, before applying the method to a real dataset. We make use of data obtained from an intensive long-term observational study of UK female grey seals (Halichoerus grypus) at a single breeding colony, where three different methods are used to identify individuals within a single study: branding, tagging and photo-identification based on seal coat pattern or pelage.

Grey seal (Halichoerus grypus) pup mortality not explained by local breeding density on North Rona, Scotland

Pre-weaning pup mortality of grey seals Halichoerus grypus in a breeding colony (North Rona, Scotland, U.K.) was examined in relation to the density of local adult females over two breeding seasons. Accurate, fine-scale maps of daily seal locations within a GIS database permitted the extraction of measures of female density determined at a range of spatial scales from 5 to 50 m around each pup on the date of death. Adult densities around live pups provided the null distribution of adult densities, which were tested against the distribution of densities where deaths occurred. Pre-weaning pup mortality at this colony was not related to any of our density measures. The proportion of pups dying each day showed no seasonal trends. Analysis of topographic features from a sub-meter digital terrain model for locations where deaths occurred suggested that there were fewer mortalities in prime pupping habitats. This could have been a direct effect of habitat or the relative quality of mothers in these habitats. The distance separating mothers and pups tended to be greater on the periphery of the colony compared to more central areas. Pups in the peripheral areas may be more exposed to predation by gulls.

Variation in female grey seal (Halichoerus grypus) reproductive performance correlates to proactive-reactive behavioural types.

Consistent individual differences (CIDs) in behaviour, indicative of behavioural types or personalities, have been shown in taxa ranging from Cnidaria to Mammalia. However, despite numerous theoretical explanations there remains limited empirical evidence for selective mechanisms that maintain such variation within natural populations. We examined behavioural types and fitness proxies in wild female grey seals at the North Rona breeding colony. Experiments in 2009 and 2010 employed a remotely-controlled vehicle to deliver a novel auditory stimulus to females to elicit changes in pup-checking behaviour. Mothers tested twice during lactation exhibited highly repeatable individual pup-checking rates within and across breeding seasons. Observations of undisturbed mothers (i.e. experiencing no disturbance from conspecifics or experimental test) also revealed CIDs in pup-checking behaviour. However, there was no correlation between an individuals’ pup-checking rate during undisturbed observations with the rate in response to the auditory test, indicating plasticity across situations. The extent to which individuals changed rates of pup-checking from undisturbed to disturbed conditions revealed a continuum of behavioural types from proactive females, who maintained a similar rate throughout, to reactive females, who increased pup-checking markedly in response to the test. Variation in maternal expenditure (daily mass loss rate) was greater among more reactive mothers than proactive mothers. Consequently pups of more reactive mothers had more varied growth rates centred around the long-term population mean. These patterns could not be accounted for by other measured covariates as behavioural type was unrelated to a mother’s prior experience, degree of inter-annual site fidelity, physical characteristics of their pupping habitat, pup sex or pup activity. These findings are consistent with the hypothesis that variation in behavioural types is maintained by spatial and temporal environmental variation combined with limits to phenotype-environment matching.

Rapid changes in Atlantic grey seal milk from birth to weaning - immune factors and indicators of metabolic strain

True seals have the shortest lactation periods of any group of placental mammal. Most are capital breeders that undergo short, intense lactations, during which they fast while transferring substantial proportions of their body reserves to their pups, which they then abruptly wean. Milk was collected from Atlantic grey seals (Halichoerus grypus) periodically from birth until near weaning. Milk protein profiles matured within 24 hours or less, indicating the most rapid transition from colostrum to mature phase lactation yet observed. There was an unexpected persistence of immunoglobulin G almost until weaning, potentially indicating prolonged trans-intestinal transfer of IgG. Among components of innate immune protection were found fucosyllactose and siallylactose that are thought to impede colonisation by pathogens and encourage an appropriate milk-digestive and protective gut microbiome. These oligosaccharides decreased from early lactation to almost undetectable levels by weaning. Taurine levels were initially high, then fell, possibly indicative of taurine dependency in seals, and progressive depletion of maternal reserves. Metabolites that signal changes in the mother’s metabolism of fats, such as nicotinamide and derivatives, rose from virtual absence, and acetylcarnitines fell. It is therefore possible that indicators of maternal metabolic strain exist that signal the imminence of weaning.

Selective blubber fatty acid mobilization in lactating gray seals (Halichoerus grypus).

During negative energy balance periods, fatty acids (FAs) are mobilized to cover the metabolic demands of the body. FAs from adipose tissue are selectively mobilized according to their carbon length (CL) and number of double bonds (DBs); however, studies in vivo have focused only on fasting and nonlactating animals. During lactation, UK gray seals fast for 18 d, mobilizing a large amount of lipid from blubber to sustain their own metabolic demands and the nutritional requirements of pups. We investigated FA mobilization in individual gray seal mothers from two UK colonies sampled in 2005 and 2006. Linear mixed-effects models were used to examine to what extent the mobilization observed from FAs in blubber can be explained as a function of FAs’ CL and number of DBs. FAs were mobilized according to their structure, such that for a given CL, mobilization increased with the number of DBs, and for a given number of DBs, mobilization decreased as CL increased. This pattern of selective mobilization was very similar between colonies, although the relative amounts of component FAs in blubber at early lactation were different between them. FAs, which are considered crucial to pup development, were mobilized more than predicted by the model. This suggests that selective mobilization of FAs is not related solely to the physicochemical characteristics of the FAs but also to the needs of a growing pup.

Conspecific recognition and aggression reduction to familiars in newly weaned, socially plastic mammals

Recognising conspecifics and behaving appropriately towards them is a crucial ability for many species. Grey seals (Halichoerus grypus) show varying capabilities in this regard: mother-pup recognition has been demonstrated in some geographical populations but is absent in others, yet there is evidence that individuals aggregate with prior associates. The recognition capabilities of newly weaned grey seal pups were investigated using class recognition trials within the habituation/dishabituation paradigm. Trials took place in pens, using pairs of individuals that either had previously cohabited (familiar) or that had never met before (stranger). Frequencies of olfactory and visual investigative behaviours (‘checks’) and aggressive interactions were recorded during trials. Familiar individuals recognised each other: paired strangers showed significantly more checks and aggressive interactions than were seen in trials pairing familiars. Oxytocin concentrations in post-trial plasma samples were analysed to investigate the underlying physiology modulating recognition abilities; however, no significant differences were detected between familiar or stranger trials. This study demonstrates that at a young age, grey seals can recognise individuals they have previously encountered. Recognition abilities in this species have adaptive value by allowing the reduction of costly aggressive interactions between familiar conspecifics, which is often cited as the first step towards the evolution of sociality in a species. This study is the first with wild subjects to find conspecific recognition abilities in a pinniped species outside of reproductive contexts. It demonstrates that even largely solitary species can be capable of recognition and pro-social behaviours that benefit them during times when they must aggregate.