DOI: 10.6084/m9.figshare.23563686
title: Multiple morphophysiological responses of a tropical frog to urbanization conform to the pace-of-life syndrome

description: Here, we analysed a suite of physiological variables that reflect whole organism health, reproduction, metabolic and circulatory physiology, and immune function in *Leptodactylus podicipinus*. We collected these data in Campo Grande, a city in central Brazil, and in an Ecological Field Station in the Pantanal. Specifically, we tested how leukocyte profile, red blood cell morphometrics, and germ cell composition, as well as somatic indices and erythrocyte nuclear abnormalities differ throughout adult life span between urban and rural populations. We used Phenotypic Trajectory Analysis to test the effect of age and site on each of multivariate datasets; and a GLM to test the effect of site and age on nuclear abnormalities. Somatic indices, erythrocyte nuclear abnormalities, red cell morphometrics, and leukocyte profile differed between populations, but less so for germ cell composition. We found a large effect of site on nuclear abnormalities, with urban frogs having twice more abnormalities than rural ones. Our results suggest that urban frogs have a faster pace of life, but this is not homogeneous across phenotypic compartments. <br> We divided the data in three files, as described below. Each file describes measurements either taken at the individual, tissue, or cell level. <br> ## Description of the data and file structure <br> This is a freeform section for you to describe how the data are structured and how a potential consumer might use them. Be as descriptive as necessary. Keep in mind that users of your data might be new to the field and unfamiliar with common terminology, metrics, etc. <br> Describe relationships between data files, missing data codes, other abbreviations used. Be as descriptive as possible. <br> **Germ_cells.csv** <br> Contains cell density of each type of germ cell and the interstitial space (columns) for each individual frog (rows). Cells plus insterstitium add up to 100%. There is also information about the age class (1-2 years, 3-4 years, and 5-6 years) and site (BEP = rural site; CG = urban site) in which frogs where collected. <br> For the evaluation of germ cells, we analysed 10 photos per animal in the 40x objective. Structural volumetric density was used to estimate the percentage of each cell type at spermatogenesis stages: spermatogonia, spermatocytes, spermatids, and loose spermatozoa in the lumen. Analysis was conducted using a grid with 252 intersections in Image J software. The density of each structure was calculated following the formula: Dve = (Ip x 100/252), where Ip are the positive intersections for the structure and 252 is the total number of intersections in the image. <br> **Planilha_individual.csv** <br> This file contains all data taken at the individual level, which include leukocyte profile, somatic indices, and erythrocyte nuclear abnormalities. There is also information about the age class (1-2 years, 3-4 years, and 5-6 years) and site (BEP = rural site; CG = urban site) in which frogs where collected. LAGs refers to the continuous Lines of Arrested Growth that were then grouped together in three age classes. <br> Body size (Snout-Vent Length, mm) of animals was measured with a digital calliper. Whole animals and individual organs were weighed on a precision scale (to the nearest 0.001 g) and the testes, spleen, and cardiac ventricle were removed to calculate somatic indices. Splenosomatic (ESI) and gonadosomatic (GSI) indexes and ventricle relative mass (RVM) were calculated as the ratio of organ weight/animal weight*100. We used the Scaled Mass Index (SMI) to estimate the body condition of animals. <br> We sampled blood from 21 individuals (10 from rural and 11 from urban sites). To describe the leukocyte profile of the two populations, we counted 100 leukocytes in each blood smear to establish the percentage of each leukocyte found in frogs (differential leukocyte count): lymphocyte, basophils, neutrophils, monocytes, thrombocyte, and eosinophil. To count the total number of nuclear abnormalities, we evaluated 1,000 erythrocyte nuclei per animal recording all types of abnormalities <br> **Red_cell_morphometrics.csv** <br> We sampled blood from 21 individuals (10 from rural and 11 from urban sites). One blood smear per individual was made immediately after blood collection and stained with May-Grunwald-Giemsa-Wright. Then, we measured cell and nuclear volume, cell and nuclear area, nucleus:cytoplasm ratio, and shape factor in 50 cells per animal (5 cells * 10 photos). There is also information about the age class (1-2 years, 3-4 years, and 5-6 years) and site (BEP = rural site; CG = urban site) in which frogs where collected. <br> <br> ## Code <br> All data analysis workflow is described in the following associated R Markdown dynamic document. R and R packages versions are described in the document below. <br> `Analysis.Rmd` and `Analysis.html`