Survival costs and benefits of reproduction: A register-based study in 20th century Estonia.

Patterns of individual variation in lifespan and senescence depend on the associations between parental survival and reproductive rates. We studied the associations between parity and survival among 579,271 Estonians born between 1905 and 1945 and in a cohort with a completed lifespan born in 1905-1927. For this cohort, selection for increased lifespan operated on both sexes, but it was stronger in men than in women. However, the median lifespan increased between the subsequent cohorts in women but stagnated in men. Selection for longer lifespan was caused by the below-average lifespan of individuals with no or single offspring. Despite a general positive selection for lifespan, survival costs of reproduction were also detected among a relatively small proportion of individuals with high parities, as mothers of two and fathers of two and three children had the highest median lifespans. Fathers of more than six children had better survival than fathers of few children in their reproductive age, but this association reversed after age 70. The reversal of this association between survival and parity at old age indicates that relative mortality risks between those with lower versus higher parities change across ages, as predicted by the antagonistic pleiotropy theory of aging.

The effect of environmental pollution on gene expression of seabirds: A review.

One of the biggest challenges for ecotoxicologists is to detect harmful effects of contaminants on individual organisms before they have caused significant harm to natural populations. One possible approach for discovering sub-lethal, negative health effects of pollutants is to study gene expression, to identify metabolic pathways and physiological processes affected by contaminants. Seabirds are essential components of ecosystems but highly threatened by environmental changes. Being at the top of the food chain and exhibiting a slow pace of life, they are highly exposed to contaminants and to their ultimate impacts on populations. Here we provide an overview of the currently available seabird-related gene expression studies in the context of environmental pollution. We show that studies conducted, so far, mainly focus on a small selection of xenobiotic metabolism genes, often using lethal sampling protocols, while the greater promise of gene expression studies for wild species may lie in non-invasive procedures focusing on a wider range of physiological processes. However, as whole genome approaches might still be too expensive for large-scale assessments, we also bring out the most promising candidate biomarker genes for future studies. Based on the biased geographical representativeness of the current literature, we suggest expanding studies to temperate and tropical latitudes and urban environments. Also, as links with fitness traits are very rare in the current literature, but would be highly relevant for regulatory purposes, we point to an urgent need for establishing long-term monitoring programs in seabirds that would link pollutant exposure and gene expression to fitness traits.

Correlations between oxidative DNA damage and formation of hepatic tumours in two flatfish species from contaminated environments.

Many species in aquatic environments face increased exposure to oncogenic pollution due to anthropogenic environmental change which can lead to higher cancer prevalence. The mechanistic relationship connecting environmental pollution and cancer is multi-factorial and poorly understood, and the specific mechanisms are so far still uncharacterized. One potential mediator between pollutant exposure and cancer is oxidative damage to DNA. We conducted a study in the field with two flatfish species, European flounder (Platichthys flesus L.) and common dab (Limanda limanda L.) with overlapping distribution and similar ecological niche, to investigate if the link between oncogenic pollutants and cancer described in ecotoxicological literature could be mediated by oxidative DNA damage. This was not the case for flounders as neither polycyclic aromatic hydrocarbon (PAH) bile metabolites nor metallic trace element concentrations were related to oxidative DNA damage measurements. However, dabs with higher PAH concentrations did exhibit increased oxidative damage. High oxidative DNA damage also did not predict neoplasm occurrence, rather, healthy individuals tended to have higher oxidative damage measurements compared to fishes with pre-neoplastic tumours. Our analyses showed that flounders had lower concentrations of PAH bile metabolites, suggesting that compared to dab this species is less exposed or better at eliminating these contaminants.

Mothers of small-bodied children and fathers of vigorous sons live longer.

Life-history traits (traits directly related to survival and reproduction) co-evolve and materialize through physiology and behavior. Accordingly, lifespan can be hypothesized as a potentially informative marker of life-history speed that subsumes the impact of diverse morphometric and behavioral traits. We examined associations between parental longevity and various anthropometric traits in a sample of 4,000-11,000 Estonian children in the middle of the 20th century. The offspring phenotype was used as a proxy measure of parental genotype, so that covariation between offspring traits and parental longevity (defined as belonging to the 90th percentile of lifespan) could be used to characterize the aggregation between longevity and anthropometric traits. We predicted that larger linear dimensions of offspring associate with increased parental longevity and that testosterone-dependent traits associate with reduced paternal longevity. Twelve of 16 offspring traits were associated with mothers' longevity, while three traits (rate of sexual maturation of daughters and grip strength and lung capacity of sons) robustly predicted fathers' longevity. Contrary to predictions, mothers of children with small bodily dimensions lived longer, and paternal longevity was not linearly associated with their children's body size (or testosterone-related traits). Our study thus failed to find evidence that high somatic investment into brain and body growth clusters with a long lifespan across generations, and/or that such associations can be detected on the basis of inter-generational phenotypic correlations.

Comparative study of the evolution of cancer gene duplications across fish.

Comparative studies of cancer-related genes not only provide novel information about their evolution and function but also an understanding of cancer as a driving force in biological systems and species' life histories. So far, these studies have focused on mammals. Here, we provide the first comparative study of cancer-related gene copy number variation in fish. Fishes are a paraphyletic group whose last common ancestor is also an ancestor of the tetrapods, and accordingly, their tumour suppression mechanisms should include most of the mammalian mechanisms and also reveal novel (but potentially phylogenetically older) previously undetected mechanisms. We have matched the sequenced genomes of 65 fish species from the Ensemble database with the cancer gene information from the COSMIC database. By calculating the number of gene copies across species using the Ensembl CAFE data (providing species trees for gene copy number counts), we used a less resource-demanding method for homolog identification. Our analysis demonstrates a masked relationship between cancer-related gene copy number variation (CNV) and maximum lifespan in fish species, suggesting that a higher number of copies of tumour suppressor genes lengthens and the number of copies of oncogenes shortens lifespan. Based on the positive correlation between the number of copies of tumour suppressors and oncogenes, we show which species have more tumour suppressors in relation to oncogenes. It could be suggested that these species have stronger genetic defences against oncogenic processes. Fish studies could be a largely unexplored treasure trove for understanding the evolution and ecology of cancer, providing novel insights into the study of cancer and tumour suppression, in addition to fish evolution, life-history trade-offs, and ecology.

Effects of environmentally relevant concentrations of microplastics on amphipods.

Lack of microplastics (MP) toxicity studies involving environmentally relevant concentrations and exposure times is concerning. Here we analyzed the potential adverse effects of low density polyethylene (LDPE) MP at environmentally relevant concentration in sub-chronic exposure to two amphipods Gmelinoides fasciatus and Gammarus lacustris, species that naturally compete with each other for their habitats. 14-day exposure to 2 µg/L (8 particles/L corresponding to low exposure) and 2 mg/L (∼8400 particles/L, corresponding to high exposure) of 53-100 µm LDPE MP were used to assess ingestion and egestion of MP, evaluate its effects on amphipod mortality, swimming ability and oxidative stress level. Both amphipod species were effectively ingesting and egesting LDPE MP. On the average, 0.8 and 2.5 MP particles were identified in the intestines of each amphipod exposed to 2 µg/L and 2 mg/L LDPE MP, respectively. Therefore, intestinal MP after 14-day exposure did not fully reflect the differences in LDPE MP exposure concentrations. Increased mortality of both amphipods was observed at 2 mg/L LDPE MP and in case of G. lacustris also at 2 µg/L exposure. The effect of LDPE on swimming activity was observed only in case of G. fasciatus. Oxidative stress marker enzymes SOD, GPx and reduced glutathione GSH varied according to amphipod species and LDPE MP concentration. In general G. lacustris was more sensitive towards LDPE MP induced oxidative stress. Overall, the results suggested that in MP polluted environments, G. lacustris may lose its already naturally low competitiveness and become overcompeted by other more resistant species. The fact that in the sub-chronic foodborne exposure to environmentally relevant and higher LDPE MP concentrations all the observed toxicological endpoints were affected refers to the potential of MP to affect and disrupt aquatic communities in the longer perspective.

Sexually antagonistic selection on educational attainment and body size in Estonian children.

Natural selection is a key mechanism of evolution, which results from the differential reproduction of phenotypes. We describe fecundity selection at different parity transitions on 15 anthropometric traits and educational attainment in Estonian children sampled in the middle of 20th century. The direction of selection on educational attainment and bodily traits was sexually antagonistic, and it occurred via different parity transitions in boys and girls. Compared to boys with primary education, obtaining tertiary education was associated with 3.5 times and secondary education two times higher odds of becoming a father. Transition to motherhood was not related to educational attainment, while education above primary was associated with lower odds (OR = 0.5-0.7) to progression to parities above one and two. Selection on anthropometric traits occurred almost exclusively via childlessness in boys, while among the girls, most of the traits that were associated with becoming a mother were additionally associated with a transition from one child to higher parities. Male (but not female) fitness was thus primarily determined by traits related to mating success. Selection favored stronger and larger boys and smaller girls. Selection on girls favored some traits that associate with perceived femininity, while other feminine traits were selected against.

Parental age does not influence offspring telomeres during early life in common gulls (Larus canus).

Parental age can affect offspring telomere length through heritable and epigenetic-like effects, but at what stage during development these effects are established is not well known. To address this, we conducted a cross-fostering experiment in common gulls (Larus canus) that enabled us distinguish between pre- and post-natal parental age effects on offspring telomere length. Whole clutches were exchanged after clutch completion within and between parental age classes (young and old) and blood samples were collected from chicks at hatching and during the fastest growth phase (11 days later) to measure telomeres. Neither the ages of the natal nor the foster parents predicted the telomere length or the change in telomere lengths of their chicks. Telomere length (TL) was repeatable within chicks, but increased across development (repeatability = 0.55, intraclass correlation coefficient within sampling events 0.934). Telomere length and the change in telomere length were not predicted by post-natal growth rate. Taken together, these findings suggest that in common gulls, telomere length during early life is not influenced by parental age or growth rate, which may indicate that protective mechanisms buffer telomeres from external conditions during development in this relatively long-lived species.

Anthropometrics of Estonian children in relation to family disruption: Thrifty phenotype and Trivers-Willard effects.

BACKGROUND AND OBJECTIVES: The thrifty phenotype hypothesis proposes that at resource limitation, the growth of some organs/tissues is selectively spared to preserve more critical ones, such as the brain or lungs. The Trivers-Willard hypothesis (TWH) predicts that boys are more vulnerable in the case of resource limitation than girls. Both hypotheses were tested in children from disrupted families, differing in the extent of deprivation/adversities imposed on them. METHODOLOGY: In a retrospective cohort study in the mid-20th century Estonia (Juhan Aul's database), different types of orphans and children of divorced parents (treatment groups; n = 106-1401) were compared with children from bi-parental families (control groups; n = 2548-8648) so that children from treatment groups were matched with control children on the basis of sex, age, year of birth, urban versus rural origin and socioeconomic position. RESULTS: Children in orphanages suffered strong growth suppression, best explained by psychosocial deprivation. Their feet were on average 0.5 SD shorter than the feet of the controls, followed by height, leg/torso ratio and cranial volume that differed from controls by ca 0.4 SD. Weight difference was 0.2 SD units, while body mass index did not differ from controls. The growth of boys and girls in orphanages was suppressed to the same extent. Boys whose mothers were dead were relatively smaller and less masculine than girls from such families. Fathers' absence was unrelated to growth suppression. Sons of divorced parents had broader shoulders than boys whose fathers were dead. CONCLUSIONS AND IMPLICATIONS: Prediction of TWH about the greater vulnerability of male growth may hold under some conditions but not universally. Predictions of the thrifty phenotype hypothesis were partly supported: trunk growth was spared at the expense of leg growth; however, no evidence for brain sparing was found. Comparison of children of divorced versus dead fathers may appear useful for indirect assessment of sexual selection on offspring quality.Lay Summary: Boys and girls in orphanages suffered similarly strong growth suppression, best explained by psychosocial deprivation. Boys whose mothers were dead were relatively smaller and less masculine than girls from such families. The occurrence of sex-specific associations between family structure and children's growth depends on the type of family disruption.

Seagrass beds reveal high abundance of microplastic in sediments: A case study in the Baltic Sea.

Microplastic (MPL) contamination in the marine environment is extensively studied yet little is known about the extent of MPL abundance in seagrass beds. The aim of this study was to evaluate MPL accumulation in coastal seagrass (Zostera marina) beds in the Baltic Sea, Estonia. Surface water was sampled by pumping using 40 µm plankton net, and sediments by trowel. MPL was extracted with NaCl, identified by microscopy and ATR-FTIR on selected samples. Surface water in the seagrass beds had 0.04-1.2 (median 0.14) MPL/L, similar to other areas of the Baltic Sea. Sediments had 0-1817 (median 208) MPL/kg (dwt), much higher than previously recorded from adjacent unvegetated and offshore sediments, thereby suggesting a strong ability of the sediments in seagrass beds to retain MPL. Of identified MPL, blue fibres were dominant in both the sampled media. Sediment characterization showed a correlation between MPL counts with poorly sorted sediments.

Exposure to artificial light at night alters innate immune response in wild great tit nestlings.

The large-scale impact of urbanization on wildlife is rather well documented; however, the mechanisms underlying the effects of urban environments on animal physiology and behaviour are still poorly understood. Here, we focused on one major urban pollutant - artificial light at night (ALAN) - and its effects on the capacity to mount an innate immune response in wild great tit (Parus major) nestlings. Exposure to ALAN alters circadian rhythms of physiological processes, by disrupting the nocturnal production of the hormone melatonin. Nestlings were exposed to a light source emitting 3 lx for seven consecutive nights. Subsequently, nestlings were immune challenged with a lipopolysaccharide injection, and we measured haptoglobin and nitric oxide levels pre- and post-injection. Both haptoglobin and nitric oxide are important markers for innate immune function. We found that ALAN exposure altered the innate immune response, with nestlings exposed to ALAN having lower haptoglobin and higher nitric oxide levels after the immune challenge compared with dark-night nestlings. Unexpectedly, nitric oxide levels were overall lower after the immune challenge than before. These effects were probably mediated by melatonin, as ALAN-treated birds had on average 49% lower melatonin levels than the dark-night birds. ALAN exposure did not have any clear effects on nestling growth. This study provides a potential physiological mechanism underlying the documented differences in immune function between urban and rural birds observed in other studies. Moreover, it gives evidence that ALAN exposure affects nestling physiology, potentially causing long-term effects on physiology and behaviour, which ultimately can affect their fitness.

Linking pollution and cancer in aquatic environments: A review.

Due to the interconnectedness of aquatic ecosystems through the highly effective marine and atmospheric transport routes, all aquatic ecosystems are potentially vulnerable to pollution. Whilst links between pollution and increased mortality of wild animals have now been firmly established, the next steps should be to focus on specific physiological pathways and pathologies that link pollution to wildlife health deterioration. One of the pollution-induced pathologies that should be at the centre of attention in ecological and evolutionary research is cancer, as anthropogenic contamination has resulted in a rapid increase of oncogenic substances in natural habitats. Whilst wildlife cancer research is an emerging research topic, systematic reviews of the many case studies published over the recent decades are scarce. This research direction would (1) provide a better understanding of the physiological mechanisms connecting anthropogenic pollution to oncogenic processes in non-model organisms (reducing the current bias towards human and lab-animal studies in cancer research), and (2) allow us to better predict the vulnerability of different wild populations to oncogenic contamination. This article combines the information available within the scientific literature about cancer occurrences in aquatic and semi-aquatic species. For the first aim, we use available knowledge from aquatic species to suggest physiological mechanisms that link pollution and cancer, including main metabolic detoxification pathways, oxidative damage effects, infections, and changes to the microbiome. For the second aim, we determine which types of aquatic animals are more vulnerable to pollution-induced cancer, which types of pollution are mainly associated with cancer in aquatic ecosystems, and which types of cancer pollution causes. We also discuss the role of migration in exposing aquatic and semi-aquatic animals to different oncogenic pollutants. Finally, we suggest novel research avenues, including experimental approaches, analysis of the effects of pollutant cocktails and long-term chronic exposure to lower levels of pollutants, and the use of already published databases of gene expression levels in animals from differently polluted habitats.

Age-dependent expression of cancer-related genes in a long-lived seabird.

Studies of model animals like mice and rats have led to great advances in our understanding of the process of tumorigenesis, but this line of study has less to offer for understanding the mechanisms of cancer resistance. Increasing the diversity of nonmodel species from the perspective of molecular mechanisms of natural cancer resistance can lead to new insights into the evolution of protective mechanisms against neoplastic processes and to a wider understanding of natural cancer defense mechanisms. Such knowledge could then eventually be harnessed for the development of human cancer therapies. We suggest here that seabirds are promising, albeit currently completely ignored candidates for studying cancer defense mechanisms, as they have a longer maximum life span than expected from their body size and rates of energy metabolism and may have thus evolved mechanisms to limit neoplasia progression, especially at older ages. We here apply a novel, intraspecific approach of comparing old and young seabirds for improving our understanding of aging and neoplastic processes in natural settings. We used the long-lived common gulls (Larus canus) for studying the age-related pattern of expression of cancer-related genes, based on transcriptome analysis and databases of orthologues of human cancer genes. The analysis of differently expressed cancer-related genes between young and old gulls indicated that similarly to humans, age is potentially affecting cancer risk in this species. Out of eleven differentially expressed cancer-related genes between the groups, three were likely artifactually linked to cancer. The remaining eight were downregulated in old gulls compared to young ones. The downregulation of five of them could be interpreted as a mechanism suppressing neoplasia risk and three as increasing the risk. Based on these results, we suggest that old gulls differ from young ones both from the aspect of cancer susceptibility and tumor suppression at the genetic level.

Corticosterone levels correlate in wild-grown and lab-grown feathers in greenfinches (Carduelis chloris) and predict behaviour and survival in captivity.

Level of corticosterone (CORT), which is a predominant glucocorticoid in birds, has become the main indicator for assessing the stress level of birds in ecological studies. Feather corticosterone (CORTf) provides information about corticosterone levels during feather growth, however, the underlying causes of individual variation of CORTf between individuals and individual persistency of CORTf are not yet fully understood. Therefore, this study addresses individual consistency in CORTf and the association of variation in CORTf with behaviour that results in damage to tail feathers. We studied relations between CORTf, plasma CORT, and behaviour in wild-caught male greenfinches in captivity. CORTf in wild-grown feathers correlated positively with CORTf in lab-grown feathers. CORTf levels were about 20% lower in lab-grown feathers than in those grown in the wild. Four birds that died in captivity had significantly higher average CORTf levels in wild-grown feathers than the survivors. Plasma CORT levels of two measurements taken in the lab seven days apart correlated positively, however, no correlations between plasma CORT and CORTf were found. In order to study the link between CORTf and behaviour, the extent of tail damage from flapping against cage bar was assessed. Contrary to our prediction, birds with higher CORTf had less tail damage. This study adds to the evidence that CORTf levels can be considered as informative markers of some persistent component of individual phenotypic quality that can predict survival under standardized laboratory conditions.

Morphometric traits predict educational attainment independently of socioeconomic background.

BACKGROUND: Aim of this study is to describe the relationship between anthropometric traits and educational attainment among Estonian schoolchildren born between 1937 and 1962. We asked whether height, cranial volume and face width (a testosterone-dependent trait), measured in childhood predict later educational attainment independently of each other, family socioeconomic position (SEP) and sex. Associations between morphometric traits and education and their interactions with biosocial variables are of scholarly importance because higher education is nearly universally associated with low fertility in women, and often with high fertility in men. Hence, morphometric traits associated with educational attainment are targeted by natural selection and describing the exact nature of these associations is relevant for understanding the current patterns of evolution of human body size. METHODS: Data on morphometric measurements and family background of 11,032 Estonian schoolchildren measured between seven and 19 years of age were obtained from the study performed by Juhan Aul between 1956 and 1969. Ordinal logistic regression was used for testing the effects of morphometric traits, biosocial variables and their interaction on the cumulative probability of obtaining education beyond primary level. RESULTS: Of biosocial variables, family SEP was the most important determinant of educational attainment, followed by the sex, rural vs urban origin and the number of siblings. No significant interactions with morphometric traits were detected, i.e., within each category of SEP, rural vs urban origin and sex, taller children and those with larger heads and relatively narrower faces were more likely to proceed to secondary and/or tertiary education. The effect of height on education was independent of cranial volume, indicating that taller children did not obtain more educations because their brains were larger than those of shorter children; height per se was important. CONCLUSIONS: Our main finding - that adjusting for other morphometric traits and biosocial variables, morphometric traits still robustly predicted educational attainment, is relevant for understanding the current patterns of evolution of human body size. Our findings suggest that fecundity selection acting on educational attainment could be partly responsible for the concurrent selection for smaller stature and cranial volume in women and opposite trends in men.

Increasing predation risk with light reduces speed, exploration and visit duration of invasive ship rats (Rattus rattus).

Exploiting predation cues to deter pests remains an untapped management tool for conservationists. We examined foraging and movement patterns of 20 wild ship rats (Rattus rattus) within a large, outdoor 'U maze' that was either illuminated or dark to assess if light (an indirect predation cue) could deter rodents from ecologically vulnerable locations. Light did not alter rats' foraging behaviour (latency to approach seed tray, visits to seed tray, time per visit to seed tray, total foraging duration, foraging rate) within the experimental resource patch but three of seven movement behaviours were significantly impaired (53% fewer visits to the maze, 70% less exploration within the maze, 40% slower movement within the maze). The total time males spent exposed to illumination also declined by 45 minutes per night, unlike females. Individual visits tended to be longer under illumination, but the latency to visit and the latency to cross through the U maze were unaffected by illumination. Elevating predation risk with illumination may be a useful pest management technique for reducing ship rat activity, particularly in island ecosystems where controlling mammalian predators is paramount to preserving biodiversity.

Uropygial gland size: a marker of phenotypic quality that shows no senescence in a long-lived seabird.

Studies of senescence in the wild have traditionally focused on traits like survival or fecundity. Although efforts to measure other salient phenotypic traits and markers of relevant physiological processes are rapidly increasing, traits related to self-maintenance remain understudied in the context of aging. Uropygial or preen gland is a holocrine gland, exclusive to birds, directly linked to self-maintenance of the quality of plumage. We measured the size of uropygial glands of common gulls (Larus canus) in a cross sectional manner in order to test whether it shows the similar age-related decline as reproductive traits previously recorded in this species. Gulls with larger glands started breeding earlier in the season, indicating that gland size is a marker of individual phenotypic quality. We found a senescent decline in the onset of breeding and the size of white wing patches, a sexually dimorphic ornamental trait, while in contrast, preen gland increased with advancing age. This finding supports the view of life-history theory that in long-lived species whose lifetime reproductive success depends heavily on lifespan, self-maintenance is prioritized over reproduction. Altogether our results support the concept that senescence in the wild can be asynchronous for traits related to maintenance versus reproduction.

Simple and noninvasive method for assessment of digestive efficiency: Validation of fecal steatocrit in greenfinch coccidiosis model.

Animals' capability to absorb energy and nutrients from food poses a major internal constraint that affects the amount of resources available for allocation to maintenance, growth, signaling, and reproduction. Intestinal surface is the largest area of contact between immune system and microbial antigens; gut thus appears the main arena where trade-offs between immune function and other components of fitness arise. Assessment of the integrity of digestive machinery should therefore be of high priority in ecophysiological research. Traditional methods of digestive physiology, however, appear unsuitable for most ecological applications due to lethality or complexity of the procedure.Here, we test the reliability of a simple, cheap, and noninvasive procedure, an acid steatocrit that assesses fat content in feces. It is based on centrifugation of a fecal sample, diluted in acid medium, in hematocrit capillary tube and quantifying the percentage of fat in fecal matter. The method has been previously validated in humans and mice; here, we apply it for the first time in birds.When applied to captive wild-caught greenfinches, the method showed reasonable internal consistency (rs = 0.71 for steatocrit values, sampled from the same fecal aliquot in duplicate but processed separately). Individual steatocrit values were significantly repeatable in time in different intervals from eight to at least 20 days (rs = 0.32-0.49). The relationship between intestinal health and steatocrit values was tested by experimental manipulations. Medication against coccidiosis (a naturally pervasive intestinal infection) reduced, and experimental infection with heterologous coccidian strains increased steatocrit. Individual changes in steatocrit correlated negatively with changes of two markers of nutritional state-plasma triglyceride levels and body mass.Findings of this study suggest that steatocrit has a wide application potential as a marker of intestinal health in ecophysiological research. In particular, we see the perspective of this method for increasingly popular immunoecological research, conservation medicine, and studies of animal coloration.

Variation in the Markers of Nutritional and Oxidative State in a Long-Lived Seabird: Associations with Age and Longevity.

Age-related declines in life-history traits have been widely observed in free-living animals. Several theories link senescence to oxidative stress. The aim of this study was to measure several widely used markers of oxidative and nutritional state in a long-lived seabird, the common gull (Larus canus), in order to assess the suitability of these markers for describing deterioration in physiological condition associated with chronological age and survival. Associations with longevity and individual consistency of these parameters over the years (repeatability) were also assessed. Senescence in fitness parameters was observed during the study period: in females, laying date and clutch mass were related to bird age in a curvilinear manner, with middle-aged birds breeding earlier and laying heavier eggs. The only parameter associated with aging processes was glutathione concentration in erythrocytes, which was lower in female birds with longer life spans. Of indexes of nutritional state, plasma triglyceride concentration showed a between-individual increase with age, suggesting selective mortality of birds with low levels. Additionally, total plasma protein levels of individual males increased with age. The mostly negative results of this study hint that the commonly used parameters of physiological condition and oxidative state used in this study do not adequately reflect an individual's long-term health condition. Alternatively, it is possible that in common gulls, senescence occurs in reproductive mechanisms but not in mechanisms responsible for maintaining an organism's redox balance, consistent with the idea that different aspects of an organism's physiological condition age at different rates. Significant interannual repeatability was detected in three plasma constituents-carotenoids, uric acid, and total protein-all of which can possibly be linked to variation in dietary habits.

Profile of whole blood gene expression following immune stimulation in a wild passerine.

BACKGROUND: Immunoecology aims to explain variation among hosts in the strength and efficacy of immunological defences in natural populations. This requires development of biomarkers of the activation of the immune system so that they can be collected non-lethally and sampled from small amounts of easily obtainable tissue. We used transcriptome profiling in wild greenfinches (Carduelis chloris) to detect whole blood transcripts that most profoundly indicate upregulation of antimicrobial defences during acute phase response. The more general aim of this study was to obtain a functional annotation of a substantial portion of the greenfinch transcriptome that would enable to gain access to more specific genomic tools in subsequent studies. The birds received either bacterial lipopolysaccharide or saline injections and RNA-seq transcriptional profiling was performed 12 h after treatment to provide initial functional annotation of the transcriptome and assess whole blood response to immune stimulation. RESULTS: A total of 66,084 transcripts were obtained from de novo Trinty assembly, out of which 23,153 could be functionally annotated. Only 1,911 of these were significantly upregulated or downregulated. The manipulation caused marked upregulation of several transcripts related to immune activation. These included avian-specific antimicrobial agents avidin and gallinacin, but also some more general host response genes, such as serum amyloid A protein, lymphocyte antigen 75 and copper-transporting ATPase 1. However, links with avian immunity for most differentially regulated transcripts remained rather hypothetical, as a large set of differentially expressed transcripts lacked functional annotation. CONCLUSIONS: This appears to be the first large scale transcriptional profiling of immune function in passerine birds. The transcriptomic data obtained suggest novel markers for the assessment of the immunological state of wild passerines. Characterizing the function of those possible novel infection markers would assist future vertebrate genome annotation. The extensive sequence information collected enables to identify possible target and housekeeping genes needed to gain access to more specific genomic tools in future studies.