Consequences of measurement error in qPCR telomere data:A simulation study

The qPCR method provides an inexpensive, rapid method for estimating relative telomere length across a set of biological samples. Like all laboratory methods, it involves some degree of measurement error. The estimation of relative telomere length is done subjecting the actual measurements made (the Cq values for telomere and a control gene) to non-linear transformations and combining them into a ratio (the TS ratio). Here, we use computer simulations, supported by mathematical analysis, to explore how errors in measurement affect qPCR estimates of relative telomere length, both in cross-sectional and longitudinal data. We show that errors introduced at the level of Cq values are magnified when the TS ratio is calculated. If the errors at the Cq level are normally distributed and independent of true telomere length, those in the TS ratio are positively skewed and proportional to true telomere length. The repeatability of the TS ratio declines sharply with increasing error in measurement of the Cq values for telomere and/or control gene. In simulated longitudinal data, measurement error alone can produce a pattern of low correlation between successive measures of relative telomere length, coupled with a strong negative dependency of the rate of change on initial relative telomere length. Our results illustrate the importance of reducing measurement error: a small increase in error in Cq values can have large consequences for the power and interpretability of qPCR estimates of relative telomere length. The findings also illustrate the importance of characterising the measurement error in each dataset-coefficients of variation are generally unhelpful, and researchers should report standard deviations of Cq values and/or repeatabilities of TS ratios-and allowing for the known effects of measurement error when interpreting patterns of TS ratio change over time

Telomere length measurement by qPCR in birds is affected by storage method of blood samples

Given the potential role of telomeres as biomarkers of individual health and ageing, there is an increasing interest in studying telomere dynamics in a wider range of taxa in the fields of ecology and evolutionary biology. Measuring telomere length across the lifespan in wild animal systems is essential for testing these hypotheses, and may be aided by archived blood samples collected as part of longitudinal field studies. However, sample collection, storage, and DNA extraction methods may influence telomere length measurement, and it may, therefore, be difficult to balance consistency in sampling protocol with making the most of available samples. We used two complementary approaches to examine the impacts of sample storage method on measurements of relative telomere length (RTL) by qPCR, particularly focusing on FTA (Flinders Technology Associates) cards as a long-term storage solution. We used blood samples from wandering albatrosses collected over 14 years and stored in three different ways (n = 179), and also blood samples from captive zebra finches (n = 30) that were each stored using three different methods. Sample storage method influenced RTL in both studies, and samples on FTA cards had significantly shorter RTL measurements. There was no significant correlation between RTL measured in zebra finch blood on FTA cards and the same samples stored either as frozen whole blood or as extracted DNA. These results highlight the importance of consistency of sampling protocol, particularly in the context of long-term field studies, and suggest that FTA cards should not be used as a long-term storage solution to measure RTL without validation

Understanding diversity in telomere dynamics

No abstract available

Data from: Maternal longevity and offspring sex in wild ungulates

In species with sexual size dimorphism, offspring of the larger sex usually have greater energy requirements and may lead to greater fitness costs for parents. The effects of offspring sex on maternal longevity, however, have only been tested in humans. Human studies produced mixed results and considerable debate mainly due to the difficulty of distinguishing the effects of sexual dimorphism from sociocultural factors. To advance this debate, we examined how the relative number of sons influenced maternal longevity in four species of free-living ungulates (Soay sheep Ovis aries; bighorn sheep, Ovis canadensis; red deer, Cervus elaphus; mountain goat, Oreamnos americanus), with high male-biased sexual size dimorphism but without complicating sociocultural variables. We found no evidence for a higher cumulative cost of sons than of daughters on maternal longevity. For a given number of offspring, most females with many sons in all four populations lived longer than females with few sons. The higher cost of sons over daughters on maternal lifespan reported by some human studies may be the exception rather than the rule in long-lived iteroparous species

Dataframes used for analyses in the paper

The first file includes data related to longevity and reproductive success in early life of females that survived beyond the onset of actuarial senescence. This data was used to produce Table 2 and Figure 1. The second file includes data related to longevity and weaning success in early life of females that survived beyond the onset of actuarial senescence. This data was used to produce Table 3 and Figure 1. The third file includes data related to longevity and lifetime reproductive success of adult females. This data was used to produce Table 4 and Figure 2. The fourth file includes data related to longevity and lifetime weaning success of adult females. This data was used to produce Table 5 and Figure 2

Plant community‐specific greening patterns predict population size increases in a temperate herbivore

Climate change‐driven impacts on vegetation productivity have been shown to drive mammalian herbivore population dynamics in Arctic and alpine environments. However, there is less evidence for temperate systems. To address this, we examined the contribution of increasing plant biomass in different vegetation communities (measured by NDVI, normalised difference vegetation index) and winter weather on the observed long‐term upward trend in the population of the Soay sheep of Hirta, St Kilda, UK. We found that biomass had increased in all vegetation communities present and increased the fastest in vegetation types preferred by the sheep. Specifically, those communities with high specific leaf area and Ellenberg's N, low leaf dry matter content. Peak summer NDVI and either winter average wind speed or winter North Atlantic Oscillation data added to the variance explained by a simple density dependence model of yearly sheep population growth rates. The highest explanatory power was found for preferred vegetation types including maritime cliff communities dominated by Plantago species, but also for both inaccessible (Rumex acetosa‐dominated) or unpreferred (Eriophorum vaginatum‐ or Sphagnum‐dominated) communities where seasonal variation more closely reflects productivity due to minimal grazing. Although the climate is getting windier and wetter, it is also getting warmer allowing increased plant productivity and this appears to be behind the long‐term increases in the Soay sheep population. Our study indicates that analysing key vegetation communities may reveal these links better than using landscape‐level averages, and that oceanic‐temperate systems may show similar climate‐driven herbivore population trends to those reported in Arctic and alpine systems.</jats:p

Dryad statement - Brown Mol Ecol

General information about the data used in this paper, including Soay Sheep Project contact information

FEC_lambs

Data for associations between SNPs and FEC in lamb

ANA_yearlings

Data for associations between SNPs and ANA in yearling

FEC_yearlings

Data for associations between SNPs and FEC in yearling