https://doi.org/10.5061/dryad.tmpg4f553

These data comprise 13 files and one annotated R script entitled 'Asota MS Analysis R 250923'. These data report the alkaloidal compounds extracted from Asota moths (Erebidae: Aganainae). We studied two species of Asota (A. eusemioides and A. heliconia) across three species of Ficus (F. septica, F. pachyrrhachis, and F. hispidioides) in a rearing experiment outlined in detail in Fontanilla et al. (2022). We collected moths at different developmental stages (caterpillars and adults) as well as their frass and leaves from their host plants. All samples were analysed using UHPLC-MS/MS and in total we found 43 alkaloidal compounds and using multivariate data analysis we explored the occurrence of compounds across sample types (including across adult body parts). Univariate analyses were used to explore compound occurrence based on trends discovered in these exploratory analyses. Below we report the experimental details needed to interpret these data.

The first part of our experiment was aimed at rearing caterpillars on each host to adulthood; this allowed accurate identification of species and developmental stage, and confirmation that body size increased predictably across instars (Dyar 1890). We selected two individual trees from each host species (n = 6) and attempted to rear five individual first instar caterpillars (collected from the appropriate host species in the surrounding forest) to adulthood (n = 30). Individual caterpillars were placed on a single branch of the Ficus species from which they were sampled and enclosed in a fine breathable mesh for rearing. Larvae were reared on young, fully expanded, and healthy leaves until their last instar, and frass was regularly removed (at least daily) to keep conditions as clean as possible. Caterpillars were followed from 13/02/2018 until 01/03/18, by which time 21 out of 30
larvae had entered pupation. Eleven larvae were reared to adulthood and stored at − 20 °C before being freeze dried. Caterpillar body length was measured to the nearest 0.1 cm on a daily basis. We also collected 10 leaves from a separate branch of the same tree before placing the caterpillars
(n = 48; leaves from one tree were excluded and one tree was sampled for eight leaves). These leaves were placed in an ice box and transferred to a − 20 °C freezer before being freeze dried at the Binatang Research Centre.

For our main experiment selected 12 trees of each host species. For each tree, we aimed to place 15 individual caterpillars of the appropriate Asota species on individual branches covered by mesh bags. Due to the local availability of first instar caterpillars, however, we ended up with 29 caterpillars on F. septica, 32 on F. hispidioides, and 31 on F. pachyrrhachis, for a total of 92 caterpillars. As with the first part, caterpillars were allowed to feed only on young and healthy leaves.Once caterpillars had molted into the last instar, ca. one half (n = 44) were freeze dried and collected as larval samples while the other half (n = 48) were reared to adults. The former were starved for six hours prior to freeze drying to ensure that there was no contamination from plant metabolites. Caterpillars in the rearing treatment were allowed to feed for 24 hr after their final molt before being placed into pots with fresh leaves. Their frass was collected every 45 min for four and a half hours. The caterpillars were then returned to their host plants to pupate naturally, and adult moths were freeze dried after emergence. A total of 17 caterpillars from the rearing treatment died in the last instar and were removed; 15 of the 17 deaths were due to parasitoid attack. We also removed four caterpillars and one adult (and their frass samples) due to possible degradation, to give a
total of 70 insect samples. Our aim was to collect six frass samples per caterpillar, for a total of 30 frass samples per tree. Because some samples were pooled in the chemical analysis due to low mass, we ended up with 170 frass samples instead of 180. In these cases, samples from the same individual—but not across individuals—were pooled. Frass was frozen immediately upon collection, as were the leaves upon which caterpillars had fed, and were later freeze dried. As the leaves on which the caterpillars fed were in generally poor condition and potentially influenced by caterpillar feeding itself, we again collected and freeze dried 10 leaves from a separate branch of the same tree to serve as baseline comparison.After accounting for excluded specimens, a total of 362 samples were used for subsequent analyses. All materials were sent to the University of Turku for chemical analyses.

Our data consists of 13 files, each file is used in the analyses outlined in the paper and in the R script 'Asota MS Analysis R 250923'. Here we provide a summary of each file but link to sections of the R script for clarity.

Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_2_Comm.txt
=> Community data (compound data by sample) used in multivariate analyses (## 07 PCA, pPCA and pCCA based on tissue type ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_2_Env.txt
=> Environmental data (moth species, host species and sample type) used in multivariate analyses (## 07 PCA, pPCA and pCCA based on tissue type ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_2_Individuals.txt
=> Individual data (a list that links individual moths to samples, e.g., a moth can be analysed as an adult for three body parts or as a caterpillar producing separate frass samples) used in multivariate analyses (## 07 PCA, pPCA and pCCA based on tissue type ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_Adults_Only_2_Comm.txt
=> Community data (compound data by adult body part) used in multivariate analyses (## 09 PCA, pPCA and pCCA based on adult body parts ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_Adults_Only_2_Env.txt
=> Environmental data (moth species, host species and adult body part) used in multivariate analyses (## 09 PCA, pPCA and pCCA based on adult body parts ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_Adults_Only_2_Individuals.txt
=> Individual data (a list that links individual moths to samples, e.g., a moth can be analysed as an adult for three body parts) used in multivariate analyses (## 09 PCA, pPCA and pCCA based on adult body parts ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_Singletons_2_Comm.txt
=> Community data (compound data by sample) used in multivariate analyses using means per individual (## 08 PCA and CCA based on tissue type with no repeated measures across samples ####)
Asota Data 230720_ALKALOIDS_DEADCATS_070222_Means_NO_DEADCATS_Singletons_2_Env.txt
=> Environmental data (moth species, host species and sample type) used in multivariate analyses using means per individual (## 08 PCA and CCA based on tissue type with no repeated measures across samples ####)
Asota Data 230720_ALKALOIDS_DEADCATS_NO_DEADCATS_080222_2.txt
=> Data used for univariate analyses of alkaloid occurence across tissue types (05 Zero inflated linear mixed model DEADCATS 070222)
Asota Data 230720_ALKALOIDS_DEADCATS_NO_DEADCATS_080222_Adults_Only_2.txt
=> Data used for univariate analyses of alkaloid occurence across adult body parts (06 Zero inflated linear mixed model adults only DEADCATS 070222)
Asota Data 230720_ALKALOIDS_DEADCATS_NO_DEADCATS_080222_Adults_Only_Reduced_2.txt
=>Data including only 'compounds of interest for adult body parts (03 Box plot for Adult Body Parts)
Instar Data.txt
=> Data used for linear mixed models to assess size across larval instars (## 02 Instar Analysis ####)
Instar Data_Asota_eusemioides.txt
=> Data used to test for a difference in larval development of Asota eusemoides across F. hispidioides and F. pachyrrhachis (## 02 Instar Analysis ####)

Sharing/Access information

These data were collected for the Fontanilla et al. (2022) and are not published elsewhere.

Code/Software

We have provided an annotated and reproducible R script in the file 'Asota MS Analysis R 250923' deposited on Zenodo.