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.

Divorce in the socially monogamous zebra finch: Hormonal mechanisms and reproductive consequences.

Up to 80% of all bird species are socially monogamous. Divorce (switching partners) or pair disruption (due to the death of a partner) has been associated with decreased reproductive success, suggesting social monogamy is a strategy that may maximize fitness via coordination between partners. Previous studies have demonstrated the effects of divorce and pair disruption on immediate reproductive success. Here, we used a paired experimental design in the zebra finch (Taeniopygia guttata) to examine the hormonal mechanisms that modulate parental behavior and reproductive success in response to a partnership change (hereafter divorce). Specifically, we examined the effects of divorce on the avian stress hormone corticosterone (CORT) in both parents and nestlings, parental behaviors (incubation and nestling provisioning), prolactin (PRL), and reproductive success. We found that divorce resulted in delayed clutch initiation, reduced clutch mass, and an increase in nestling CORT response to a standardized stressor. These effects on reproductive investment and chick CORT response were not clearly determined by parental endocrine responses. Divorce had no effect on the level of parental CORT. PRL levels were highly correlated within a pair regardless of treatment, were negatively related to the investment that males made in incubation, and increased in experimental males as a result of pair disruption. This study demonstrates the fundamental impact which divorce has not only on reproduction, but also the physiological stress responses of offspring and suggests that in socially monogamous animals the maintenance of a stable partnership over time could be advantageous for long term fitness.

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 animal's 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.

Field Evaluation of Melolure, a Formate Analogue of Cuelure, and Reassessment of Fruit Fly Species Trapped in Sydney, New South Wales, Australia.

In Australia, tephritids are usually attracted to either cuelure or methyl eugenol. Methyl eugenol is a very effective lure, but cuelure is less effective likely due to low volatility. A new formate analogue of cuelure, melolure, has increased volatility, resulting in improved efficacy with the melon fruit fly, Bactrocera cucurbitae Coquillett. We tested the efficacy of melolure with fruit fly species in Sydney as part of the National Exotic Fruit Fly Monitoring programme. This monitoring programme has 71 trap sites across Sydney, with each trap site comprising separate Lynfield traps containing either cuelure, methyl eugenol, or capilure lure. In 2008, an additional Lynfield trap with melolure plugs was added to seven sites. In 2009 and 2010, an additional Lynfield trap with melolure wicks was added to 11 trap sites and traps were monitored fortnightly for 2 yr. Capture rates for melolure traps were similar to cuelure traps for Dacus absonifacies (May) and Dacus aequalis (Coquillet), but melolure traps consistently caught fewer Bactrocera tryoni (Froggatt) than cuelure traps. However, trap sites with both a cuelure and melolure traps had increased capture rates for D. absonifacies and D. aequalis, and a marginally significant increase for B. tryoni. Melolure plugs were less effective than melolure wicks, but this effect may be related to lure concentration. The broader Bactrocera group species were attracted more to cuelure than melolure while the Dacus group species were attracted more to melolure than cuelure. There is no benefit in switching from cuelure to melolure to monitor B. tryoni, the most important fruit fly pest in Australia.

Raspberry ketone feeding makes Queensland fruit fly, Bactrocera tryoni (Froggatt), more vulnerable to desiccation but not starvation.

BACKGROUND: Queensland fruit fly (Qfly) males exhibit accelerated sexual maturation when their diet is supplemented with raspberry ketone (RK) for 48 h following emergence, which is beneficial for sterile insect technique operation. The present study tests whether RK supplementation makes Qfly more vulnerable to starvation or desiccation. RESULTS: Flies were fed for 48 h with a yeast hydrolysate and sugar diet (1:3) that contained 0% RK (control), 1.25% RK (low dose) or 5% RK (high dose) to test subsequent vulnerability to starvation and desiccation. RK feeding decreased body weight and water content in males and increased lipid levels in both sexes before exposure to any stress treatment. Under nutritional stress, flies fed the low RK dose, but not the high RK dose, had higher survival than controls. Under desiccation stress, flies fed both the low and high RK doses had lower survival than the controls. Body weight, water content and lipid reserves at death were all affected by RK dose when under nutritional stress, but not when under desiccation stress. In the absence of stress, body weight at death was higher than controls in flies provided with the high RK dose and lipids were lower than controls in flies provided with the low RK dose. CONCLUSION: Feeding with RK makes Qflies more vulnerable to desiccation but not starvation. In most conditions, it is expected that the disadvantage of increased desiccation vulnerability would be outweighed by the benefits of accelerated sexual maturation in RK-fed young adult Qflies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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 fly's 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.

Geometry of compensatory feeding and water consumption in Drosophila melanogaster.

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.

Eating and wheel running across the estrous cycle in rat lines selectively bred on a taste phenotype.

The Occidental Low- and High-Saccharin-Consuming rats (respectively, LoS and HiS) have been selectively bred for decades to study the relationship between taste and behaviors in and beyond the ingestive domain. Whether the saccharin phenotype is associated with behavioral periodicities tied to reproductive status is not known. Here we describe for the first time variation across the estrous cycle in chow intake and wheel running by LoS and HiS rats. This study also shed light on why rats, humans, and some other mammals eat less and become more active as fertility increases. Wheel running increases when eating is reduced through restricted chow access, more so in LoS rats than in HiS rats (Dess et al., 2000). If the decrease in food intake from diestrus through estrus causes the increase in running (Eat Less â†’ Run More hypothesis, ELRM), then the running peak should follow the eating nadir and be greater in LoS rats. Bayesian cyclic regression showed that estrous cycles were shorter in LoS rats than in HiS rats; implications are discussed. Contrary to ELRM, the running peak did not follow the eating nadir, and cycle amplitude did not distinguish LoS rats from HiS rats. These results indirectly support the No Time To Eat hypothesis (Fessler, 2003), according to which the periovulatory eating nadir and running peak reflect fitness-enhancing consequences of shifts away from eating and toward mating as fertility increases.

Towards powerful experimental and statistical approaches to study intraindividual variability in labile traits.

There is a long-standing interest in behavioural ecology, exploring the causes and correlates of consistent individual differences in mean behavioural traits ('personality') and the response to the environment ('plasticity'). Recently, it has been observed that individuals also consistently differ in their residual intraindividual variability (rIIV). This variation will probably have broad biological and methodological implications to the study of trait variation in labile traits, such as behaviour and physiology, though we currently need studies to quantify variation in rIIV, using more standardized and powerful methodology. Focusing on activity rates in guppies (Poecilia reticulata), we provide a model example, from sampling design to data analysis, in how to quantify rIIV in labile traits. Building on the doubly hierarchical generalized linear model recently used to quantify individual differences in rIIV, we extend the model to evaluate the covariance between individual mean values and their rIIV. After accounting for time-related change in behaviour, our guppies substantially differed in rIIV, and it was the active individuals that tended to be more consistent (lower rIIV). We provide annotated data analysis code to implement these complex models, and discuss how to further generalize the model to evaluate covariances with other aspects of phenotypic variation.

Mating Reverses Actuarial Aging in Female Queensland Fruit Flies.

Animals that have a long pre-reproductive adult stage often employ mechanisms that minimize aging over this period in order to preserve reproductive lifespan. In a remarkable exception, one tephritid fruit fly exhibits substantial pre-reproductive aging but then mitigates this aging during a diet-dependent transition to the reproductive stage, after which life expectancy matches that of newly emerged flies. Here, we ascertain the role of nutrients, sexual maturation and mating in mitigation of previous aging in female Queensland fruit flies. Flies were provided one of three diets: 'sugar', 'essential', or 'yeast-sugar'. Essential diet contained sugar and micronutrients found in yeast but lacked maturation-enabling protein. At days 20 and 30, a subset of flies on the sugar diet were switched to essential or yeast-sugar diet, and some yeast-sugar fed flies were mated 10 days later. Complete mitigation of actuarial aging was only observed in flies that were switched to a yeast-sugar diet and mated, indicating that mating is key. Identifying the physiological processes associated with mating promise novel insights into repair mechanisms for aging.

Cyclic changes in cortisol across the estrous cycle in parous and nulliparous Asian elephants.

In the context of reproduction, glucocorticoids (GCs) are generally considered to have negative effects. However, in well-studied model species, GCs fluctuate predictability across the estrous cycles, and short-term increases promote healthy ovarian function. Reproductive challenges have plagued captive elephant populations, which are not currently self-sustaining. Efforts to understand reproductive dysfunction in elephants have focused on the suppressive effects of cortisol, but the potential permissive or stimulatory effects of cortisol are unknown. In this study, we provide a detailed examination of cortisol patterns across the estrous cycle in Asian elephants (Elephas maximus). Time series analysis was used to analyze cortisol and progesterone data for a total of 73 cycles from eight females. We also compared cortisol profiles between females that successfully conceived and females that failed to conceive despite repeated mating attempts. Our results revealed that cortisol fluctuates predictably across the estrous cycle, with a peak during the second half of the follicular phase followed by low levels throughout the luteal phase. Furthermore, this pattern was significantly altered in nulliparous females; cortisol concentrations did not decline during the luteal phase to the same extent as in parous females. This study highlights the complexity of cortisol signaling and suggests future directions for understanding the role of cortisol in reproductive dysfunction.

Automated locomotor activity monitoring as a quality control assay for mass-reared tephritid flies.

BACKGROUND: The Sterile Insect Technique (SIT) requires vast numbers of consistently high quality insects to be produced over long periods. Quality control (QC) procedures are critical to effective SIT, both providing quality assurance and warning of operational deficiencies. We here present a potential new QC assay for mass rearing of Queensland fruit flies (Bactrocera tryoni Froggatt) for SIT; locomotor activity monitoring. We investigated whether automated locomotor activity monitors (LAMs) that simply detect how often a fly passes an infrared sensor in a glass tube might provide similar insights but with much greater economy. RESULTS: Activity levels were generally lower for females than for males, and declined over five days in the monitor for both sexes. Female activity levels were not affected by irradiation, but males irradiated at 60 or 70 Gy had reduced activity levels compared with unirradiated controls. We also found some evidence that mild heat shock of pupae results in adults with reduced activity. CONCLUSION: LAM offers a convenient, effective and economical assay to probe such changes.

Diet quality mediates activity patterns in adult Queensland fruit fly (Bactrocera tryoni).

Studies linking resource acquisition and trait expression have traditionally treated nutritional resources as a single currency, but recent research has shown that trait expression can depend as much on diet quality (nutrient composition) as on diet quantity (calories). Here, we investigate the role of nutrient composition and diet concentration on activity levels of adult Queensland fruit flies (Bactrocera tryoni Froggatt: Tephritidae). Male and female flies were fed diets that varied in the proportion of protein and carbohydrate as well as total amounts of protein and carbohydrate. Daily activity levels were then quantified using locomotor activity monitors during both light and dark phases. During the light phase, both sexes increased the proportion of time spent active and their rate of activity as diets became more carbohydrate-rich and concentrated. In contrast, during the dark phase, nutrient composition and concentration had no effect on the proportion of time spent active for either sex, although when active during the dark phase, activity rates were higher for flies fed more carbohydrate-rich and concentrated diets. Overall, nutritional composition of the diet affected activity levels to a greater extent than the total energetic content of the diet.

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.

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.

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.