DOI: 10.5061/dryad.ttdz08m47
title: Dataset for: Warmer ambient temperatures reduce protein intake by a mammalian folivore

description: The interplay between ambient temperature and nutrition in wild herbivores
 is frequently overlooked, despite the fundamental importance of food. We
 tested whether different ambient temperatures (10˚C, 18˚C, and 26˚C)
 influenced the intake of protein by a marsupial herbivore, the common
 brushtail possum (Trichosurus vulpecula). At each temperature, possums
 were offered a choice of two foods containing different amounts of protein
 (57% vs 8%) for one week. Animals mixed a diet with a lower proportion of
 protein to non-protein (P:NP, 0.20) when held at 26˚C compared to that at
 both 10˚C and 18˚C (0.22). Since detoxification of plant secondary
 metabolites imposes a protein cost on animals, we then studied whether
 addition of the monoterpene 1,8-cineole to the food changed the effect of
 ambient temperature (10˚C and 26˚C) on food choice. Cineole reduced food
 intake but also removed the effect of temperature on P:NP ratio and
 instead animals opted for a diet with higher P:NP (0.19 with cineole vs
 0.15 without cineole). These experiments show the proportion of P:NP
 chosen by animals is influenced by ambient temperature and by plant
 secondary metabolites. Protein is critical for reproductive success in
 this species and reduced protein intake caused by high ambient
 temperatures may limit the viability of some populations in the future.
description: These data were collected
 using twelve adult male common brushtail possums
 (<em>Trichosurus</em> <em>vulpecula</em>) which
 were caught in wire cage traps on the Australian National University
 campus (Canberra, 35.2809° S, 149.1300° E) in November 2016. The possums were housed inside constant
 temperature rooms (at 10, 18, or 26 °C ± 2°C) with a 12:12 h light cycle
 and offered artificial diets prepared daily (composition is described in
 the uploaded dataset). Food was offered at 1630 h and refusals were
 collected at 0900 h daily. Dry matter intake (DMI) of the two diets was
 measured by drying a subsample of the food offered at 80 °C and
 subtracting the dry mass of all food refusals. In addition, we calculated
 the daily grams of protein (P) and “non-protein” (NP; sum of fats and
 non-structural carbohydrates) ingested across 7 days for each possum for
 each temperature. Possums were weighed weekly. 
 The first experiment tested
 whether possums balanced macronutrients differently depending on the
 ambient temperature. The second tested how the addition of a PSM, cineole
 influenced the effect of ambient temperature on macronutrient
 balancing. Experiment 1: Macronutrient choice at different
 ambient temperatures Possums were randomly allocated into three
 groups of four, and each group was kept at a constant temperature of
 either 10, 18 or 26 ± 2°C for seven days. The possums were then rotated between the
 temperature treatments for two more rounds of the experiment, using a
 randomized crossover design so the number of observations on each
 treatment was n=12. Experiment 2: The effects of ambient temperature
 and cineole on macronutrient choice
 Possums were randomly
 allocated into two groups. One group was kept at 26 ± 2°C for seven
 nights, while the other was kept at 10 ± 2°C. Possums were offered the two
 diets differing in macronutrient composition without addition of any
 1,8-cinoele for the first six nights. DMI and diet preferences were
 measured on the fifth and sixth nights and the mean was taken as an
 estimate of intake of the basal diets. On the seventh night 1,8-cineole
 was added to both diets at a concentration of 2.45% WM (~7% DM) and DMI
 and diet preferences were measured again over two
 nights.
description: # Macronutrient\_Temperature Experiments 1. Title of Dataset: Warmer
 ambient temperatures reduce protein intake by a mammalian folivore [Access
 this dataset on Dryad](https://doi.org/10.5061/dryad.ttdz08m47) 2.
 Principal investigator: Phillipa K Beale Australian National University
 phillipa.beale@gmail.com Co-authors: Ben D Moore, William J Foley, Karen J
 Marsh ## Summary of dataset collection: These data were collected using
 twelve adult male common brushtail possums (*Trichosurus* *vulpecula*)
 which were caught on the Australian National University campus (Canberra,
 35.2809° S, 149.1300° E) in November 2016. The possums were housed inside
 constant temperature rooms (at 10, 18, or 26 °C ± 2°C) with a 12:12 h
 light cycle and offered artificial diets prepared daily (composition is
 described in the uploaded dataset). Food was offered at 1630 h and
 refusals were collected at 0900 h daily. Dry matter intake (DMI) of the
 two diets was measured by drying a subsample of the food offered at 80 °C
 and subtracting the dry mass of all food refusals. In addition, we
 calculated the daily grams of protein (P) and “non-protein” (NP; sum of
 fats and non-structural carbohydrates) ingested across 7 days for each
 possum for each temperature. Possums were weighed weekly. * Experiment 1:
 Macronutrient choice at different ambient temperatures Possums were
 randomly allocated into three groups of four, and each group was kept at a
 constant temperature of either 10, 18 or 26 ± 2°C for seven days. The
 possums were then rotated between the temperature treatments for two more
 rounds of the experiment, using a randomized crossover design so the
 number of observations on each treatment was n=12. * Experiment 2: The
 effects of ambient temperature and 1,8-cineole on macronutrient choice
 Possums were randomly allocated into two groups. One group was kept at 26
 ± 2°C for seven nights, while the other was kept at 10 ± 2°C. Possums were
 offered the two diets differing in macronutrient composition for the first
 six nights. DMI and diet preferences were measured on the fifth and sixth
 nights and the mean was taken as an estimate of intake of the basal diets.
 On the seventh night 1,8-cineole was added to both diets at a
 concentration of 2.45% WM (\~7% DM) and DMI and diet preferences were
 measured again over two nights. * pretrial: the data set also includes
 results from a short acclimation period between the two experiments to
 introduce cineole to the possums less suddenly. The results from this
 period were not used in the final analysis of the data for publication. ##
 Abbreviations used: * HP = high protein diet * LP = low protein diet * DMI
 = dry matter intake * WM = Wet matter * O/N = overnight ## Description of
 the data and file structure Number of files: 1 Number of tabs: 9
 (described below) ## Info: General information about the experiments
 including number of animals, time of food collection, and weights of
 animals in grams ### HP&LP Diets: (HP = high protein diet, LP =
 low protein diet) A description of the dietary formulation in percentage
 of WM (%WM) #### Designs: Experiment 1 has purple headers and experiment 2
 has orange headers. * Possum (individual animal identifier numbers are
 used), * Group (possums were allocated to groups and moved together), *
 Round 1 (Which temperature treatment allocated in experimental round 1), *
 Round 2 (which temperature treatment allocated in round 2), * Round 3
 (which temperature treatment allocated in round 3). #### Raw data 1: The
 raw data from experiment 1 in terms of... * exp round (first second or
 third round of temperature treatments), * day of round (how many days at
 that temperature), * date offered (the date food was given), * possum (in
 terms of individual identifier number), * temp (temperature in degrees
 celsius of the room, 10, 18 or 26) * HP offered (WM g) = how much of the
 high protein diet was offered in grams of wet matter * HP offered (DM g) =
 how much of the high protein diet was offered in grams of dry matter,
 calculated from the percentage of dry matter in dried control samples. *
 HP %DM = percentage of dry matter in the high protein diet, based on the
 control samples * HP dish (g) = the weight of the dish the high protein
 food was given in * dish + HPDM left (g) = weight in grams of the dish and
 the dried refusals from the high protein diet * HP DMI (g) = how much of
 the high protein diet was eaten by the possum in grams of dry matter. * LP
 offered (WM g) = how much low protein diet was offered in grams of wet
 matter * LP offered (DM g) = how much of the low protein diet was offered
 in grams of dry matter based on the percentage dry matter in the control
 sample * LP %DM = percentage of the low protein diet that is dry matter
 based on dried control samples * LP dish (g) = weight in grams of the dish
 the low protein diet was offered in * dish +LPDM left (g) = weight in
 grams of the dish and the dried refusals of the low protein diet * LP DMI
 = grams of dry matter intake of the low protein diet * Proportion of
 HPvsLP: proportion of high protein diet compared to low protein diet eaten
 by the possum. * total DMI = total (HP plus LP) dry matter intake CONTROLS
 1: the controls from experiment 1 which were subsamples from the diets
 offered that day, dried to calculate dry matter content of the diet. *
 Date = date that diet was offered to possums * HP WM (g) = weight of the
 wet matter in grams of the high protein diet control subsample * HP dish
 (g) = weight of the dish the sample is being dried in in grams * HP DM (g)
 = weight in grams of the high protein diet subsample once dried * HP%DM =
 percentage of the high protein diet that was dry matter * LP WM (g) =
 grams of wet matter of low protein diet being weighed * LP dish (g) =
 weight in grams of the dish that the low protein diet subsample is being
 weighed in * LP DM (g) = Weight in grams of the low protein diet subsample
 * LP%DM = percentage of the low protein diet that is dry matter * Notes
 PRETRIAL: raw data from an acclimation period between experiment 1 and
 experiment 2 not used in final analysis for publication. Headings are: *
 exp round (either half the cineole concentration used in the experiment or
 the full 7% cineole) * day of round (day number), * date offered (the date
 food was given), * possum (in terms of individual identifier number), *
 temp (temperature in degrees celsius of the room, 10, 18 or 26) * HP
 offered (WM g) = how much of the high protein diet was offered in grams of
 wet matter * HP offered (DM g) = how much of the high protein diet was
 offered in grams of dry matter, calculated from the percentage of dry
 matter in dried control samples. * HP %DM = percentage of dry matter in
 the high protein diet, based on the control samples * HP dish (g) = the
 weight of the dish the high protein food was given in * dish + HPDM left
 (g) = weight in grams of the dish and the dried refusals from the high
 protein diet * HP DMI (g) = how much of the high protein diet was eaten by
 the possum in grams of dry matter. * LP offered (WM g) = how much low
 protein diet was offered in grams of wet matter * LP offered (DM g) = how
 much of the low protein diet was offered in grams of dry matter based on
 the percentage dry matter in the control sample * LP %DM = percentage of
 the low protein diet that is dry matter based on dried control samples *
 LP dish (g) = weight in grams of the dish the low protein diet was offered
 in * dish +LPDM left (g) = weight in grams of the dish and the dried
 refusals of the low protein diet * LP DMI = grams of dry matter intake of
 the low protein diet * Proportion of HPvsLP: proportion of high protein
 diet compared to low protein diet eaten by the possum. * total DMI = total
 (HP plus LP) dry matter intake #### PRETRIAL CONTROLS: control subsamples
 used to calculate dry matter content of the diets offered in the pretrial
 * Date = date that diet was offered to possums * HP WM (g) = weight of the
 wet matter in grams of the high protein diet control subsample * HP dish
 (g) = weight of the dish the sample is being dried in in grams * HP DM (g)
 = weight in grams of the high protein diet subsample once dried * HP%DM =
 percentage of the high protein diet that was dry matter * LP WM (g) =
 grams of wet matter of low protein diet being weighed * LP dish (g) =
 weight in grams of the dish that the low protein diet subsample is being
 weighed in * LP DM (g) = Weight in grams of the low protein diet subsample
 * LP%DM = percentage of the low protein diet that is dry matter RAW DATA2:
 raw data collected in the second experiment * exp round (first or second
 round of temperature treatments), * day of round (how many days at that
 temperature), * date offered (the date food was given), * possum (in terms
 of individual identifier number), * temp (temperature in degrees celsius
 of the room, 10, 18 or 26) * psm = plant secondary metabolite treatment,
 basal = basal diet (no cineole) or cineole (basal with cineole added) * HP
 offered (WM g) = how much of the high protein diet was offered in grams of
 wet matter * HP offered (DM g) = how much of the high protein diet was
 offered in grams of dry matter, calculated from the percentage of dry
 matter in dried control samples. * HP %DM = percentage of dry matter in
 the high protein diet, based on the control samples * HP dish (g) = the
 weight of the dish the high protein food was given in * dish + HPDM left
 (g) = weight in grams of the dish and the dried refusals from the high
 protein diet * HP DMI (g) = how much of the high protein diet was eaten by
 the possum in grams of dry matter. * LP offered (WM g) = how much low
 protein diet was offered in grams of wet matter * LP offered (DM g) = how
 much of the low protein diet was offered in grams of dry matter based on
 the percentage dry matter in the control sample * LP %DM = percentage of
 the low protein diet that is dry matter based on dried control samples *
 LP dish (g) = weight in grams of the dish the low protein diet was offered
 in * dish +LPDM left (g) = weight in grams of the dish and the dried
 refusals of the low protein diet * LP DMI = grams of dry matter intake of
 the low protein diet * Proportion of HPvsLP: proportion of high protein
 diet compared to low protein diet eaten by the possum. * total DMI = total
 (HP plus LP) dry matter intake CONTROLS2 = the control subsamples used to
 calculate dry matter content in experiment 2 * Date = date that diet was
 offered to possums * HP WM (g) = weight of the wet matter in grams of the
 high protein diet control subsample * HP dish (g) = weight of the dish the
 sample is being dried in in grams * HP DM (g) = weight in grams of the
 high protein diet subsample once dried * HP%DM = percentage of the high
 protein diet that was dry matter * LP WM (g) = grams of wet matter of low
 protein diet being weighed * LP dish (g) = weight in grams of the dish
 that the low protein diet subsample is being weighed in * LP DM (g) =
 Weight in grams of the low protein diet subsample * LP%DM = percentage of
 the low protein diet that is dry matter Publication relating to this data
 set is in preparation and can be found at the following DOI once
 published. It was supported by grants from the Australian Research Council
 (DE120101263 and DP140100228) and with support from Society for
 Experimental Biology and The Company of Biologist:
 https://doi.org/10.1098/rstb.2022.0543