DOI: 10.5061/dryad.2jm63xsxr
title: Reproduction results in parallel changes of oxidative stress and immunocompetence in a wild long-living mammal - edible dormouse <em>Glis glis</em>

description: Oxidative stress (OS) and impaired immune function (IF) have been proposed
 as key physiological costs of reproduction. The relationship between IF
 and OS, particularly in long-living iteroparous species, remains
 unresolved. Here, we studied OS and IF in breeding and non-breeding
 females of edible dormice- a long-living rodent prioritizing
 self-maintenance. We tested whether free-ranging reproducing females
 experience the trade-off between breeding investments, preventing OS, and
 sustaining IF. Because of senescence, the OS markers (oxidative damage -
 ROMs, lower antioxidant capacity - OXY) should increase with age and
 reproductive activity; the IF (indexed as the level of white blood cells –
 WBC and increased neutrophil to lymphocyte ratio - N/L) should decrease
 with age and reproductive effort. WBC decreased, N/L increased, while OS
 markers remained unaffected by age. ROMs (but not OXY) and age-adjusted
 WBC were the lowest in lactating, higher in post-lactating, and the
 highest in non-breeding females. The path analysis testing the causal
 relationship between ROMs and WBC revealed that IF is more likely to
 affect OS than vice versa. Our study indicates the trade-off between
 reproductive investments and sustaining IF and demonstrates that females
 suppress IF to reduce the risk of OS and, therefore, mitigate oxidative
 costs of reproduction.
description: The population of edible dormice was studied in the central part
 of Kozienice Forest (51°30′15″N, 21°27′27″E) in central Poland. We
 monitored this population from 2013 and marked animals individually
 (RF-IDW-1, CBDZOE, Poland). In August-September 2019-2021, females were
 captured (21 lactating, 17 post-lactating, and 44 non-lactating) and
 transported to the laboratory of Forest Research Institute in Sękocin
 Stary. Blood sampling protocol was performed after ~30 hours spent in the
 animal room to reduce the stress associated with capture and transport to
 the laboratory. During the procedure (which lasted less than 3 minutes),
 animals were anaesthetised under a 2 % isoflurane and atmospheric air.
 Blood samples (~50 µL) were taken from the retro-orbital sinus by
 heparinized microhematocrit capillary tubes. To establish the leukocyte
 status, the blood smear was prepared immediately from 4µL of full blood.
 15 min after sampling~~ blood was centrifuged at 13,000 rpm for 20 min at
 4°C to separate red blood cells from plasma. Plasma was immediately stored
 at -80°C until the assays were performed.
description: # Reproduction results in parallel changes of oxidative stress and
 immunocompetence in a wild long-living mammal - edible dormouse *Glis
 glis* #### Description of the data and file structure Data are provided as
 a single CSV file, **dormice_data.csv,** with columns * **Animal_ID** (the
 individual number of each female dormouse marked with an RFID tag) *
 **Age** (age of female marked as juvenile during previous studies, data
 available only for 17 individuals, empty cells left unfilled to not
 interfere with the R code to analyse the data) * **year** (year in which
 individual was captured and included to the study) * **sex_act**
 (reproductive status of female: 1lac - lactating, 2post-lac -
 post-lactating, 3non-lac - non-lactating) * **hw** (measured head width of
 female in the broadest point - zygomatic arches with caliper to nearest
 0.1 mm) * **dROM** (measured reactive oxygen metabolites in blood plasma
 of female, by the use of the dROM assay kit (Diacron, Grosseto, Italy)) *
 **OXYadsorbent** (measured antioxidant capacity by OXY-Adsorbent Test
 (Diacron, Grosseto, Italy)) * **neutro_sum** (sum of neutrophiles per 400
 counted leukocytes, established by evaluation of blood smear prepared from
 blood of female) * **eosinoph_sum** (sum of eosinophiles per 400 counted
 leukocytes, established by evaluation of blood smear prepared from blood
 of female) * **basoph_sum** (sum of basophiles per 400 counted leukocytes,
 established by evaluation of blood smear prepared from blood of female) *
 **mono_sum** (sum of monocytes per 400 counted leukocytes, established by
 evaluation of blood smear prepared from blood of female) * **limpho_sum**
 (sum of lymphocytes per 400 counted leukocytes, established by evaluation
 of blood smear prepared from blood of female) * **n_l** (proportion of
 neutrophiles counted per 400 leukocytes to lymphocytes counted per 400
 leukocytes; index of inflammation and stress) * **wbc** (average amount of
 white blood cells counted per 10 000 of red blood cells in 10 repeats,
 established by evaluation of blood smear prepared from blood of female) *
 **mb** (body mass of female measured after capture with a resolution of
 2g) * **body_size** (body size of female calculated as common variance of
 head width and body mass, obtained in principal component analysis,
 explaining 78% of variation in body mass and size, estimated as head
 width) * **PC_2** (condition of female calculated as common variance of
 head width and body mass, obtained in principal component analysis,
 explaining 22% of variation in body mass and size, estimated as head
 width) #### Code/Software In addition to the data, we provide the code for
 reproducing analyses in **dormice_analyses.R**. This script analyzes
 dormouse physiological data by testing relationships between reproductive
 status, body size, and immune/oxidative stress markers using mixed and
 linear models, model comparisons, and path analysis. It also checks
 assumptions, estimates marginal means, tests interactions, and evaluates
 alternative causal pathways between white blood cells and oxidative
 damage.