Effects of calcium supplementation on oxidative status and oxidative damage in great tit nestlings inhabiting a metal-polluted area.

Calcium has been proposed to diminish metal toxicity by the modulation of the oxidative stress. This study explores the effects of Ca availability and metal exposure on oxidative stress biomarkers in great tit (Parus major) nestlings. Nests were supplemented with Ca (Ca-supplemented group) or not supplemented (Control group) in a metal-polluted and a background zone in SW Finland. Metal concentrations were analyzed from feces. We analyzed antioxidants (tGSH, GSH:GSSG ratio, CAT, GST, GPx, SOD), protein carbonylation and lipid peroxidation in red cells of nestlings. Ca-supplemented and fast-growing nestlings showed higher CAT activity to cope with reactive oxygen species (ROS) generated during intensive growth and metabolism. SOD and GPx (the latter not statistically significant) were more active in the polluted area, possibly reflecting higher ROS production in nestlings from this zone due to the enhanced metal exposure and smaller size. Antioxidant levels changed over the range of metal concentrations depending on the Ca levels in plasma, suggesting that higher Ca levels stimulate antioxidants and mitigate the impacts of metals. Ca supplementation may improve nestling traits and reproductive output when antioxidants are enhanced in a situation of oxidative challenge. Therefore, Ca should be considered in future studies assessing metal exposure and effects on wild birds.

Telomere length, sibling competition and development of antioxidant defense in wild house mice.

Antioxidants and telomere length are potential biomarkers for individuals' exposure and ability to cope with environmental stressors. However, intraspecific variations in antioxidant alterations due to natural, life cycle related stress, have been rarely estimated. We investigated those changes in wild-derived house mice in a longitudinal study with natural sibling competition as a stressor. Blood was used for telomere length measurements at 8-weeks age and for several selected antioxidants at 8-weeks and 6-months age. Our results show that most of the antioxidants increase during that time, indicating that antioxidant-system continues to develop after early development and sexual maturation. In addition females had higher antioxidant-levels than males. Mice with longer telomeres had also higher superoxide dismutase-activity and more glutathione than mice with shorter telomeres, meaning that long telomeres are associated with better antioxidant defense at maturation and during later life. Sibling competition at early age affected superoxide dismutase-levels at 6-months, but only in females. Females, which were lighter than the average of the litter had low superoxide dismutase -activity in later adulthood, indicating delayed negative effect of sibling competition on antioxidant defense. Our results highlight that sex and developmental stage are crucial in intraspecific comparisons of the antioxidant status and its alterations.

Telomere length and antioxidant defense associate with parasite-induced retarded growth in wild brown trout.

Early growth conditions can have profound impacts on individuals' development, growth and physiology, with subsequent long-term consequences for individuals' fitness and life expectancy. Telomere length (TL) has been suggested to indicate both individual fitness and life expectancy in wide range of species, as the telomere attrition rate at early age can be accelerated due to exposure to various stressors, including parasites and inflammatory diseases, which increase production of reactive oxygen species (ROS) and influence antioxidant (AO) levels. We investigated impacts of Tetracapsuloides bryosalmonae infection, a causative agent of proliferative kidney disease (PKD), on AO status and TL in a natural population of juvenile brown trout (Salmo trutta). The fish with higher parasite load showed more severe kidney hyperplasia, anemia and smaller body size compared to less parasitized fish. Furthermore, fish with severe PKD symptoms had lower SOD-, CAT- and GST activity than fish with milder kidney hyperplasia. However, parasite load was not directly correlated either with AOs or with TL. Smaller fish showed shorter TLs, potentially reflecting lower individual quality. The fish, which were less sensitive to parasite-induced impaired growth, quantified as parasite load-adjusted fork length, showed also longer TLs, lower GR- and GST activity and less GSHtot compared to more sensitive fish. These results provide novel knowledge about the impacts of the PKD in brown trout at the molecular level and support the idea that TL may reflect individual quality and ability to cope with parasitic infections.

Telomere damage and redox status alterations in free-living passerines exposed to metals.

Telomere length may reflect the expected life span and possibly individual quality. Environmental stressors are known to increase oxidative stress and accelerate telomere attrition: however the interactions between redox status and telomere dynamics are not fully understood. We investigated whether exposure to heavy metal pollution is associated with oxidative stress and telomere damage in two insectivorous passerines, the Great tit (Parus major) and the Pied flycatcher (Ficedula hypoleuca). We were also interested to know whether within-brood competition could influence the nestling redox status or telomere length. Breeding females and nestlings were sampled near the point pollution source and compared to birds in non-polluted control zone. We measured heavy metal concentrations, calcium, metallothioneins, telomere lengths and redox status (oxidative damage, and enzymatic and non-enzymatic antioxidants) in liver samples. Great tit nestlings in the polluted zone had significantly shorter telomeres compared to those in the unpolluted control zone. In addition, those great tit nestlings that were lighter than their average siblings, had shorter telomeres compared to the heavier ones. In pied flycatchers neither pollution nor growth stress were associated with telomere length, but adult females had significantly shorter telomeres compared to the nestlings. All the results related to redox status varied remarkably among the species and the age groups. In both species antioxidants were related to pollution. There were no significant associations between redox status and telomere length. Our results suggest that wild birds at a young age are vulnerable to pollution and growth stress induced telomere damage. Redox status seems to interact with pollution and growth, but more studies are needed to clarify the underlying physiological mechanisms of telomere attrition. Our study highlights that all the observed associations and differences between the sampling zones varied depending on the species, age, and degree of exposure to pollution.

Effects of early-life lead exposure on oxidative status and phagocytosis activity in great tits (Parus major).

Lead is a highly poisonous metal with a very long half-life, distributing throughout the body in blood and accumulating primarily in bones and kidney. We studied the short and long-term effects of early-life lead exposure on antioxidant defense and phagocytosis activity in a small passerine bird, the great tit (Parus major) by manipulating dietary lead levels of the nestlings. We had three experimental groups, exposed to environmentally relevant lead concentrations; high (4 µg/g body mass), low (1 µg/g body mass) and control (0 µg/g body mass) group. As a comparison, a great tit population breeding in the vicinity of a metal smelter was included to the experimental set-up. We measured glutathione, the ratio of reduced and oxidized glutathione, and the antioxidant enzymes: glutathione peroxidase, glutathione-S-transferase, catalase and superoxide dismutase together with protein carbonylation and phagocytosis activity to study the effects of lead on the oxidative status and immune function of birds. We found differences in enzyme activities between the study groups, but in most cases the smelter group differed from the other groups. Despite the differences observed in antioxidant enzymes, our results indicate only minor short-term effects of lead exposure on oxidative status, since either glutathione ratio or protein carbonylation were not affected by lead. Phagocytosis activity was not linked to higher lead concentrations either. Interestingly, protein carbonylation was positively associated with enzyme activities and glutathione level. Our results did not show major long-term effects of lead on the oxidative status of great tits.

No delayed behavioral and phenotypic responses to experimental early-life lead exposure in great tits (Parus major).

Early-life exposure to pollutants, such as lead, may have long-lasting consequences on health, behavior, and cognition. However, experiments on delayed effects of specific pollutants are very rare in wild animals. We experimentally exposed wild nestling great tits (Parus major) to dietary lead (high, low, or control group) in levels relevant to exposure levels of wild populations in Europe and studied delayed effects on phenotypic and behavioral traits in captivity. We also included a group of birds from a vicinity of a copper smelter, exposed to a mixture of toxic metals and altered food supply during development. This experimental setup allowed us to compare the strength of direct (exposure to lead per se) and indirect (pollution-related changes in diet) effects of pollutants. Our experimental lead treatment significantly increased lead levels in bone and feces compared with controls. However, we found no carry-over effect of early-life dietary lead on morphology, plumage coloration, or heat shock proteins. Treatment did not affect activity, exploration, neophobia, or success in learning and spatial memory task. We conclude that with the exposure levels and relatively short exposure period used, delayed effects on the measured traits were not found. However, it is important to further study other types of behavioral traits and ultimately fitness effects.

Experimental manipulation of dietary lead levels in great tit nestlings: limited effects on growth, physiology and survival.

We manipulated dietary lead (Pb) levels of nestlings in wild populations of the great tit (Parus major L) to find out if environmentally relevant Pb levels would affect some physiological biomarkers (haematocrit [HT], fecal corticosterone metabolites [CORT], heat shock proteins [HSPs], erythrocyte delta-aminolevulinic acid dehydratase activity [ALAd]), growth (body mass, wing length), phenotype (plumage coloration) or survival of nestlings. The responses to three experimental manipulation (control, low and high: 0, 1 and 4 µg/g body mass/day) are compared with those in a P. major population breeding in the vicinity of a heavy metal source, a copper smelter. Our Pb supplementation was successful in raising the fecal concentrations to the levels found in polluted environments (high: 8.0 µg/g d.w.). Despite relatively high range of exposure levels we found only few effects on growth rates or physiology. The lack of blood ALAd inhibition suggests that the circulating Pb levels were generally below the toxic level despite that marked accumulation of Pb in femur (high: 27.8 µg/g d.w.) was observed. Instead, birds in the metal polluted environment around the smelter showed decreased growth rates, lower HT, higher CORT, less colorful plumage and lower survival probabilities than any of the Pb treated groups. These effects are likely related to decreased food quality/quantity for these insectivorous birds at the smelter site. In general, the responses of nestlings to metal exposure and/or associated resource limitation were not gender specific. One of the stress proteins (HSP60), however, was more strongly induced in Pb exposed males and further studies are needed to explore if this was due to higher accumulation of Pb or higher sensitivity of males. In all, our results emphasize the importance of secondary pollution effects (e.g. via food chain disruption) on reproductive output of birds.