Benjamin G. Fanson

Nutrients, not caloric restriction, extend lifespan in Queensland fruit flies (Bactrocera tryoni).

Caloric restriction (CR) has been widely accepted as a mechanism explaining increased lifespan (LS) in organisms subjected to dietary restriction (DR), but recent studies investigating the role of nutrients have challenged the role of CR in extending longevity. Fuelling this debate is the difficulty in experimentally disentangling CR and nutrient effects due to compensatory feeding (CF) behaviour. We quantified CF by measuring the volume of solution imbibed and determined how calories and nutrients influenced LS and fecundity in unmated females of the Queensland fruit fly, Bactocera tryoni (Diptera: Tephritidae). We restricted flies to one of 28 diets varying in carbohydrate:protein (C:P) ratios and concentrations. On imbalanced diets, flies overcame dietary dilutions, consuming similar caloric intakes for most dilutions. The response surface for LS revealed that increasing C:P ratio while keeping calories constant extended LS, with the maximum LS along C:P ratio of 21:1. In general, LS was reduced as caloric intake decreased. Lifetime egg production was maximized at a C:P ratio of 3:1. When given a choice of separate sucrose and yeast solutions, each at one of five concentrations (yielding 25 choice treatments), flies regulated their nutrient intake to match C:P ratio of 3:1. Our results (i) demonstrate that CF can overcome dietary dilutions; (ii) reveal difficulties with methods presenting fixed amounts of liquid diet; (iii) illustrate the need to measure intake to account for CF in DR studies and (iv) highlight nutrients rather than CR as a dominant influence on LS.

Protein:carbohydrate ratios explain life span patterns found in Queensland fruit fly on diets varying in yeast:sugar ratios

Dietary restriction extends life span across a vast diversity of taxa, but significant challenges remain in elucidating the underlying mechanisms. Distinguishing between caloric and nutrient effects is an essential step. Recent studies with Drosophila and tephritid fruit flies have reported increased life span as dietary yeast-to-sugar ratios decreased and these effects have been attributed to changes in protein-to-carbohydrate (P:C) ratios of the diets rather than calories. However, yeast is a complex mix of macronutrients and micronutrients, and hence changes in yeast content of the diet necessarily alters other nutrients in lockstep. To explicitly test whether studies using yeast are justified in attributing results to diet protein content rather than correlated nutrients, we developed a chemically defined diet allowing manipulation of just the ratio of protein (free amino acids) to carbohydrate (sucrose) levels of diets while holding other nutrients constant. Mated, female Queensland fruit flies (Q-flies) were fed 1 of 18 diets varying in P:C ratios and diet concentration. Diet consumption, egg production, and life span were recorded for each fly. In close concordance with recent studies using yeast diets, flies had increased life span as P:C ratios decreased, and caloric restriction did not extend life span. Similarly, egg production was maximized on high P:C ratios, but lifetime egg production was maximized on intermediate P:C ratios, indicating a life history trade-off between life span and egg production rate. Finally, Q-flies adjusted their diet intake in response to P:C ratios and diet concentration. Our results substantiate recent claims that P:C ratios significantly modulate life span in flies.

Geometry of compensatory feeding and water consumption in Drosophila melanogaster

SUMMARY Feeding behaviour is an expression of an animal’s underlying nutritional strategy. The study of feeding decisions can hence delineate nutritional strategies. Studies of Drosophila melanogaster feeding behaviour have yielded conflicting accounts, and little is known about how nutrients affect feeding patterns in this important model species. Here, we conducted two experiments to characterize nutrient prioritization and regulation. In a choice experiment, we allowed female flies to self-regulate their intake of yeast, sucrose and water by supplying individual flies with three microcapillary tubes: one containing only yeast of varying concentrations, another with just sucrose of varying concentrations, and the last with just water. Flies tightly regulated yeast and sucrose to a constant ratio at the expense of excess water intake, indicating that flies prioritize macronutrient regulation over excess water consumption. To determine the relative importance of yeast and sucrose, in a no-choice experiment, we provided flies with two microcapillary tubes: the first with one of the 28 diets varying in yeast and sucrose content and the other with only water. Flies increased total water intake in relation to yeast consumption but not sucrose consumption. Additionally, flies increased diet intake as diet concentration decreased and as the ratio of sugar to yeast equalized. Using a geometric scaling approach, we found that the patterns of diet intake can be explained by flies prioritizing protein and carbohydrates equally and by the lack of substitutability between the nutrients. We conclude by illustrating how our results harmonize conflicting results in the literature once viewed in a two-dimensional diet landscape.

Cost of reproduction in the Queensland fruit fly: Y-model versus lethal protein hypothesis.

The trade-off between lifespan and reproduction is commonly explained by differential allocation of limited resources. Recent research has shown that the ratio of protein to carbohydrate (P : C) of a flys diet mediates the lifespan–reproduction trade-off, with higher P : C diets increasing egg production but decreasing lifespan. To test whether this P : C effect is because of changing allocation strategies (Y-model hypothesis) or detrimental effects of protein ingestion on lifespan (lethal protein hypothesis), we measured lifespan and egg production in Queensland fruit flies varying in reproductive status (mated, virgin and sterilized females, virgin males) that were fed one of 18 diets varying in protein and carbohydrate amounts. The Y-model predicts that for sterilized females and for males, which require little protein for reproduction, there will be no effect of P : C ratio on lifespan; the lethal protein hypothesis predicts that the effect of P : C ratio should be similar in all groups. In support of the lethal protein hypothesis, and counter to the Y-model, the P : C ratio of the ingested diets had similar effects for all groups. We conclude that the trade-off between lifespan and reproduction is mediated by the detrimental side-effects of protein ingestion on lifespan.

Additive and interactive effects of nutrient classes on longevity, reproduction, and diet consumption in the Queensland fruit fly (Bactrocera tryoni)

Insect lifespan is often closely linked to diet, and diet manipulations have been central to studies of ageing. Recent research has found that lifespan for some flies is maximised on a very low yeast diet, but once all yeast is removed, lifespan drops precipitously. Although effects of yeast availability on lifespan are commonly interpreted in terms of protein, yeast is a complex mix of nutrients and provides a rich source of vitamins, minerals and sterols. Elucidating which components of yeast are involved in this lifespan drop provides insights into more specific nutritional requirements and also provides a test for the commonplace interpretation of yeast in terms of protein. To this end, we fed Queensland fruit flies (Bactrocera tryoni) one of eight experimental diets that differed in the nutrient group(s) found in yeast that were added to sucrose: none, vitamins, minerals, amino acids, cholesterol, vitamin+mineral+cholesterol (VMC), vitamin+mineral+cholesterol+amino acids (VMCA), and yeast. We measured survival rates and egg production in single sex and mixed sex cages, as well as nutrient intake of individual flies. We found that the addition of minerals increased lifespan of both male and female flies housed in single sex cages by decreasing baseline mortality. The addition of just amino acids decreased lifespan in female flies; however, when combined with other nutrient groups found in yeast, amino acids increased lifespan by decreasing both baseline mortality and age-specific mortality. Flies on the yeast and VMCA diets were the only ones to show significant egg production. We conclude that the drop in lifespan observed when all yeast is removed is explained by missing micronutrients (vitamins, minerals and cholesterol) as well as the absence of protein in females, whereas minerals alone can explain the pattern for males. These results indicate a need for caution when interpreting effects of dietary yeast as effects of protein.

The reproductive cycle of the Tasmanian devil (Sarcophilus harrisii) and factors associated with reproductive success in captivity

Numbers of wild Tasmanian devils are declining as a result of the fatal, transmissible Devil Facial Tumor Disease. A captive insurance population program has been initiated but current captive breeding rates are sub-optimal and therefore the goal of this project was to increase our understanding of the estrous cycle of the devil and elucidate potential causes of failed male-female pairings. Temporal patterns of fecal progestagen and corticosterone metabolite concentrations were examined for females (n=41) in three categories of reproductive status (successful: viable young, n=20 estrous cycles; unsuccessful: paired with a male but no young confirmed, n=44 estrous cycles; non-mated: no access to a male during estrus, n=8 estrous cycles) but substantial differences were not found. Females were more likely to produce pouch young if pairing with the male extended into late proestrus (P<0.05), thereby decreasing the time between pairing and presumed ovulation. The interval between the end of proestrous elevation in progestagen metabolite concentrations and the beginning of the luteal phase was 7.6±2.3 days in successful females. The length of the luteal phase in successful females was 12.5±1.4 days which was not different from unsuccessful or non-mated females (P>0.05). Unsuccessful females had 1-3 estrous cycles within a single year. Successful females were predominantly wild-caught (17/19, 90%) and most produced young following the first estrous cycle of the season (18/20, 90%). Unsuccessful females were predominantly captive born (20/27, 74%) in this study. It is possible that a proportion of females that do not produce pouch young achieve conception but the timing of reproductive failure continues to be elusive in this species.

Post-teneral nutrition as an influence on reproductive development, sexual performance and longevity of Queensland fruit flies

Adult Queensland fruit flies, Bactrocera tryoni (Froggatt), require adequate post‐teneral nutrition to complete reproductive development, to perform sexually and for maximum longevity. Recent research has focussed on nutritional requirements of adults released in sterile insect technique (SIT) programmes that are used to manage these pests. Several studies have suggested benefits of providing yeast hydrolysate (YH) in addition to sugar during the 24‐ to 48‐h pre‐release holding period. Current evidence suggests that provision of YH can induce faster development, increased mating probability, longer copulations, increased sperm storage by mates, higher levels of sexual inhibition in mated females and increased longevity. We here review research on adult B. tryoni nutritional requirements, assess the potential application of this information in the context of SIT programmes and highlight future research that will help to determine whether YH, or other supplements, should be included as pre‐release treatment in B. tryoni SIT.

A review of 16 years of quality control parameters at a mass-rearing facility producing Queensland fruit fly, Bactrocera tryoni

From 1996 to 2012, the mass‐rearing facility at Camden (NSW, Australia) has been producing Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). During this time, the facility has regularly recorded fly quality parameters, creating a unique data set that provides an invaluable opportunity to evaluate the interrelationships among standard quality control (QC) parameters and test for redundant QC variables. Here, we conducted an exploratory data analysis to reveal relationships among the QC parameters. We found that pupal weight, adult lifespan, and longevity under nutritional stress (i.e., survival duration without food or water) had distinct monthly trends, suggesting that these QC parameters are sensitive to seasonal conditions. Furthermore, emergence percentage, flight ability, and adult lifespan were adversely affected by the dyeing/handling/irradiation process associated with sterile insect releases. Using a multivariate approach and controlling for monthly and yearly patterns, we showed that pupal weight and egg hatch are consistently negatively related and that percentage male and emergence rates are consistently negatively related. These results suggest that these correlation pairs measure similar quality information and hence one QC variable from each pair could be dropped. Flight ability was not strongly correlated with any of the QC variables, suggesting that this QC variable remains a useful QC metric. Finally, the longevity under nutritional stress QC appears to be fairly insensitive to QCs and we suggest that it should be replaced by the standard mortality under stress test.

One size does not fit all: Monitoring faecal glucocorticoid metabolites in marsupials.

Marsupial research, conservation, and management can benefit greatly from knowledge about glucocorticoid (GC) secretion patterns because GCs influence numerous aspects of physiology and play a crucial role in regulating an animals response to stressors. Faecal glucocorticoid metabolites (FGM) offer a non-invasive tool for tracking changes in GCs over time. To date, there are relatively few validated assays for marsupials compared with other taxa, and those that have been published generally test only one assay. However, different assays can yield very different signals of adrenal activity. The goal of this study was to compare the performance of five different enzyme immunoassays (EIAs) for monitoring adrenocortical activity via FGM in 13 marsupial species. We monitored FGM response to two types of events: biological stressors (e.g., transport, novel environment) and pharmacological stimulation (ACTH injection). For each individual animal and assay, FGM peaks were identified using the iterative baseline approach. Performance of the EIAs for each species was evaluated by determining (1) the percent of individuals with a detectable peak 0.125-4.5days post-event, and (2) the biological sensitivity of the assay as measured by strength of the post-event response relative to baseline variability (Z-score). Assays were defined as successful if they detected a peak in at least 50% of the individuals and the mean species response had a Z⩾2. By this criterion, at least one assay was successful in 10 of the 13 species, but the best-performing assay varied among species, even those species that were closely related. Furthermore, the ability to confidently assess assay performance was influenced by the experimental protocols used. We discuss the implications of our findings for biological validation studies.

Selection on Mitochondrial Variants Occurs between and within Individuals in an Expanding Invasion

Mitochondria are critical for life, yet their underlying evolutionary biology is poorly understood. In particular, little is known about interaction between two levels of evolution: between individuals and within individuals (competition between cells, mitochondria or mitochondrial DNA molecules). Rapid evolution is suspected to occur frequently in mitochondrial DNA, whose maternal inheritance predisposes advantageous mutations to sweep rapidly though populations. Rapid evolution is also predicted in response to changed selection regimes after species invasion or removal of pathogens or competitors. Here, using empirical and simulated data from a model invasive bird species, we provide the first demonstration of rapid selection on the mitochondrial genome within individuals in the wild. Further, we show differences in mitochondrial DNA copy number associated with competing genetic variants, which may provide a mechanism for selection. We provide evidence for three rarely documented phenomena: selection associated with mitochondrial DNA abundance, selection on the mitochondrial control region, and contemporary selection during invasion.