Insect Fitness Research Articles

Enhanced capacity of a leaf beetle to combat dual stress from entomopathogens and herbicides mediated by associated microbiota.

Herbicides have demonstrated their impact on insect fitness by affecting their associated microbiota or altering the virulence of entomopathogenic fungi toward insects. However, limited research has explored the implications of herbicide stress on the intricate tripartite interaction among insects, associated bacterial communities, and entomopathogens. In this study, we initially demonstrated that associated bacteria confer a leaf beetle, Plagiodera versicolora, with the capability to resist the entomopathogenic fungus Aspergillus nomius infection, a capability sustained even under herbicide glyphosate stress. Further analysis of the associated microbiota revealed a significant alteration in abundance and composition due to glyphosate treatment. The dominant bacterium, post A. nomius infection or following a combination of glyphosate treatments, exhibited strong suppressive effects on fungal growth. Additionally, glyphosate markedly inhibited the pathogenic associated bacterium Pseudomonas though it inhibited P. versicolora's immunity, ultimately enhancing the beetle's tolerance to A. nomius. In summary, our findings suggest that the leaf beetle's associated microbiota bestow an augmented resilience against the dual stressors of both the entomopathogen and glyphosate. These results provide insight into the effects of herbicide residues on interactions among insects, associated bacteria, and entomopathogenic fungi, holding significant implications for pest control and ecosystem assessment.

Extreme heat reduces host and parasite performance in a butterfly-parasite interaction.

Environmental temperature fundamentally shapes insect physiology, fitness and interactions with parasites. Differential climate warming effects on host versus parasite biology could exacerbate or inhibit parasite transmission, with far-reaching implications for pollination services, biocontrol and human health. Here, we experimentally test how controlled temperatures influence multiple components of host and parasite fitness in monarch butterflies (Danaus plexippus) and their protozoan parasites Ophryocystis elektroscirrha. Using five constant-temperature treatments spanning 18-34°C, we measured monarch development, survival, size, immune function and parasite infection status and intensity. Monarch size and survival declined sharply at the hottest temperature (34°C), as did infection probability, suggesting that extreme heat decreases both host and parasite performance. The lack of infection at 34°C was not due to greater host immunity or faster host development but could instead reflect the thermal limits of parasite invasion and within-host replication. In the context of ongoing climate change, temperature increases above current thermal maxima could reduce the fitness of both monarchs and their parasites, with lower infection rates potentially balancing negative impacts of extreme heat on future monarch abundance and distribution.

The clip cage conundrum: Assessing the interplay of confinement method and aphid genotype in fitness studies.

Behavior and fitness are important ecological traits frequently measured in insect bioassays. A common method to measure them in soft-bodied herbivorous insects involves confining individuals to plant leaves using clip cages. Although studies have previously highlighted the negative effects of clip cages on leaf physiology, little is known about the impact that using this confinement method has on insect fitness. The responses of different aphid genotypes/clones to different containment methods have not previously been investigated. Here we measured key fitness traits (intrinsic rate of natural increase, mean relative growth rate, time to reach reproductive adulthood and population doubling time) in the potato aphid, Macrosiphum euphorbiae Thomas (Hemiptera: Aphididae), when confined to plants using two methods: (1) clip cages to confine aphids to individual strawberry leaves and (2) a mesh bag to confine aphids to whole strawberry plants. Our study identified a strong negative impact on all the measured aphid fitness traits when using clip cages instead of mesh bags. We also identified genotype-specific differences in response to confinement method, where clip cage confinement differentially affected the fitness of a given aphid genotype compared to the same genotype on whole plants. These results suggest that clip cage use should be carefully considered when experiments seek to quantify insect fitness and that whole plants should be used wherever possible. Given the prevalence of clip cage use in insect bioassays, our results highlight the need for caution when interpreting the existing literature as confinement method significantly impacts aphid fitness depending on their genotype.

The cluster digging behavior of larvae confers trophic benefits to fitness in insects.

Collective behaviors efficiently impart benefits to a diversity of species ranging from bacteria to humans. Fly larvae tend to cluster and form coordinated digging groups under crowded conditions, yet understanding the rules governing this behavior is in its infancy. We primarily took advantage of the Drosophila model to investigate cooperative foraging behavior. Here, we report that Drosophila-related species and the black soldier fly have evolved a conserved strategy of cluster digging in food foraging. Subsequently, we investigated relative factors, including larval stage, population density, and food stiffness and quality, that affect the cluster digging behavior. Remarkably, oxygen supply through the posterior breathing spiracles is necessary for the organization of digging clusters. More importantly, we theoretically devise a mathematical model to accurately calculate how the cluster digging behavior expands food resources by diving depth, cross-section area, and food volume. We found that cluster digging behavior approximately increases 2.2 fold depth, 1.7-fold cross-section area, and 1.9 fold volume than control groups, respectively. Amplification of food sources significantly facilitates survival, larval development, and reproductive success of Drosophila challenged with competition for limited food resources, thereby conferring trophic benefits to fitness in insects. Overall, our findings highlight that the cluster digging behavior is a pivotal behavior for their adaptation to food scarcity, advancing a better understanding of how this cooperative behavior confers fitness benefits in the animal kingdom.

Male-specific hexokinase functions as a seminal fluid protein to enhance insect fitness

Male-specific hexokinase functions as a seminal fluid protein to enhance insect fitness

Transmission mode predicts coinfection patterns of insect-specific viruses in field populations of the Queensland fruit fly.

Insect-specific viruses (ISVs) can affect insect health and fitness, but can also interact with other insect-associated microorganisms. Despite this, ISVs are often studied in isolation from each other, in laboratory populations. Consequently, their diversity, prevalence and associations with other viruses in field populations are less known, yet these parameters are important to understanding virus epidemiology. To help address this knowledge gap, we assessed the diversity, prevalence and coinfections of three ISVs (horizontally transmitted cripavirus, biparentally transmitted sigmavirus and maternally transmitted iflavirus) in 29 field populations of Queensland fruit fly, Australia's most significant horticultural pest, in the context of their different transmission modes. We detected new virus variant diversity. In contrast to the very high virus prevalence in laboratory populations, 46.8% of 293 field flies carried one virus and 4.8% had two viruses. Cripavirus and sigmavirus occurred in all regions, while iflavirus was restricted to subtropical and tropical regions. Cripavirus was most prevalent (37.5%), followed by sigmavirus (13.7%) and iflavirus (4.4%). Cripavirus coinfected some flies with either one of the two vertically transmitted viruses. However, sigmavirus did not coinfect individuals with iflavirus. Three different modelling approaches detected negative association patterns between sigmavirus and iflavirus, consistent with the absence of such coinfections in laboratory populations. This may be linked with their maternal transmission and the ineffective paternal transmission of sigmavirus. Furthermore, we found that, unlike sigmavirus and iflavirus, cripavirus load was higher in laboratory than field flies. Laboratory and mass-rearing conditions may increase ISV prevalence and load due to increased transmission opportunities. We conclude that a combination of field and laboratory studies is needed to uncover ISV interactions and further our understanding of ISV epidemiology.

Heat stress has immediate and persistent effects on immunity and development of Tenebrio molitor

Abstract The yellow mealworm (Tenebrio molitor) is a promising insect species for mass-rearing for the production of feed and food. In mass-production systems, insects may be exposed to abiotic stressors such as heat stress as well as potentially lethal pathogens. To ensure mass-reared T. molitor populations are healthy and productive there is a need to understand both the risks, and potential benefits of heat stress, on the fitness of insects and their susceptibility to pathogens. In this study, we investigated the effects of a short (2 h) or a long (14 h) heat stress (38 °C) exposure on the susceptibility and the immune responses of T. molitor larvae exposed to a fungal pathogen (Metarhizium brunneum). Larvae were exposed to the pathogen either immediately or five days after the heat stress treatments. The development of heat stressed larvae and their offspring was also assessed. A short heat stress immediately before exposure to M. brunneum increased the survival probability of T. molitor larvae, which correlated with increased antibacterial activity in the hemolymph. The exposure of larvae to short, or long heat stresses five days before pathogen exposure did not affect their survival, despite a temporary lowered body mass gain of heat stressed larvae. However, heat stressed larvae showed decreased hemocyte concentrations when exposed to M. brunneum. We also found an increased body weight in larval offspring of females that had been exposed to a short heat stress as larvae themselves. These findings demonstrate the importance of understanding the effects of heat stress in the long-term. The beneficial effects of heat stress on pathogen susceptibility in T. molitor and the negative effects on body mass gain are only transient, whereas negative effects on immune response (hemocyte concentrations) persist over an extended period.

Temperature Sensitivity of Fitness Components across Life Cycles Drives Insect Responses to Climate Change.

AbstractThermal performance curves (TPCs) are increasingly used as a convenient approach to predict climate change impacts on ectotherms that accounts for organismal thermal sensitivity; however, directly applying TPCs to temperature data to estimate fitness has yielded contrasting predictions depending on assumptions regarding climate variability. We compare direct application of TPCs to an approach integrating TPCs for different fitness components (e.g., per capita birth rate, adult life span) across ectotherm life cycles into a population dynamic model, which we independently validated with census data and applied to hemipteran insect populations across latitude. The population model predicted that climate change will reduce insect fitness more at higher latitudes due to its effects on survival but will reduce net reproductive rate more at lower latitudes due to its effects on fecundity. Directly applying TPCs underestimated climate change impacts on fitness relative to incorporating the TPCs into the population model due to simplifying survival dynamics across the life cycle. The population model predicted that climate change will reduce mean insect density and increase population variability at higher latitudes via reduced survival, despite faster development and a longer activity period. Our study highlights the importance of considering how multiple fitness components respond to climate variability across the life cycle to better understand and anticipate the ecological consequence of climate change.

Fitness costs of resistance to insecticides in insects

The chemical application is considered one of the most crucial methods for controlling insect pests, especially in intensive farming practices. Owing to the chemical application, insect pests are exposed to toxic chemical insecticides along with other stress factors in the environment. Insects require energy and resources for survival and adaptation to cope with these conditions. Also, insects use behavioral, physiological, and genetic mechanisms to combat stressors, like new environments, which may include chemicals insecticides. Sometimes, the continuous selection pressure of insecticides is metabolically costly, which leads to resistance development through constitutive upregulation of detoxification genes and/or target-site mutations. These actions are costly and can potentially affect the biological traits, including development and reproduction parameters and other key variables that ultimately affect the overall fitness of insects. This review synthesizes published in-depth information on fitness costs induced by insecticide resistance in insect pests in the past decade. It thereby highlights the insecticides resistant to insect populations that might help design integrated pest management (IPM) programs for controlling the spread of resistant populations.

Fungicides modify pest insect fitness depending on their genotype and population

Fungicides are the most sold pesticide group, with an 8% increase in sales in Europe within the last decade. While adverse short-term fungicide effects on non-target insect species have been reported, the long-term effects and their impact on fitness are unclear. As the effects may depend on both the fungicide and the genetic background of the species, we investigated the effects of the commonly used fungicide, fluazinam, on the Colorado potato beetle's life history traits, and whether the effects were dependent on a previously characterized insecticide resistance mutation (S291G in acetylcholinesterase-2 gene) in different populations. Our findings show that fungicide exposure can have both negative and positive, long-lasting effects on beetles, depending on the parental insecticide resistance status and population. In the Belchow population, individuals carrying resistance mutation had higher survival, but they produced offspring with lower egg-hatching rates. While, in the Vermont population, fungicide exposure increased the body mass and offspring quality in the beetles carrying resistance mutation but did not affect the beetles’ survival. Our results suggest that commonly used fungicides can have both negative and positive effects on pest insects’ life-history, however, their impact may differ depending on the population and parental genetic background.

The effect of adult density on the reproductive output and hatching performance of the lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

Abstract Commercial availability of insect biomass for food and feed calls for the methodical investigation of all the complications that can deteriorate the insect farming efficiency. An appropriate stocking density is thus instrumental for the establishment of sustainable and cost effective insect farming protocols; high densities may lead to reduced insect fitness and reproduction, while low densities will result to a proportionally lower per capita yield. The present study deals with the estimation of the optimum adult density of the lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), a species with great potential as food and feed source based on its high growth performance and reproductive output. However, limited data is available so far regarding the effect of adult density on its reproductive performance. Four different adult densities, ranging from 1.1 to 8.2 cm2 per adult, were evaluated in laboratory experiments in terms of reproductive output (number of eggs) and larval emergence for a period of 45 days of continuous oviposition. According to our results, the reproduction output, expressed as the cumulative number of eggs per adult and the cumulative larval hatching rate, was not affected by the adult densities tested here. Although high hatching rates that in most of the cases exceeded 85% were observed within the initial oviposition interval, these rates were in all treatments drastically reduced to 48-56% after 30 to 36 d. High adult survival (over 93%) was observed during the trial. In this experiment, the maximum reproductive output was observed in the highest density of adults that were left to oviposit continuously for a month. Data as such provide an opportunity to further exploit A. diaperinus in high-throughput production systems.

Burrowing depth mediates the extent of metabolic suppression in a soil‐overwintering insect

Abstract Conserving energy through winter is important for the fitness of temperate insects. While insects can use buffered microhabitats, metabolic suppression or decreases in the thermal sensitivity of metabolic rate to override seasonal‐scale thermal trends, the relative importance of these strategies for limiting energy use by insects overwintering in soil remains underexplored. We used a combined laboratory, field and simulation approach to investigate the overwintering energetics of the western bean cutworm (Striacosta albicosta), a univoltine lepidopteran pest of dry beans and corn that overwinters underground as a dormant prepupa. We hypothesised that (1) the selection of thermally buffered microhabitats (i.e. deeper soil sites) reduces energy use in early autumn and late spring, and that (2) changes in the metabolic rate–temperature relationship reduce the impact of elevated temperatures on overwintering energy use. We provide evidence that during the warmest parts of winter, dormant S. albicosta prepupae that had burrowed deep benefited from a cool, stable microclimate, whereas those near the soil surface appeared to rely on deeper metabolic suppression to maintain their energy stores. Although elevated temperatures in the laboratory depleted their energy reserves, these strategies appear sufficient to limit energy drain under natural conditions in the field. We suggest that small‐scale variation in the depth of soil refuges may mediate the interaction between the risk of energy drain and changes in the metabolic rate–temperature relationship in soil‐overwintering insects. Read the free Plain Language Summary for this article on the Journal blog.

Impact of intraspecific variation in insect microbiomes on host phenotype and evolution.

Microbes can be an important source of phenotypic plasticity in insects. Insect physiology, behaviour, and ecology are influenced by individual variation in the microbial communities held within the insect gut, reproductive organs, bacteriome, and other tissues. It is becoming increasingly clear how important the insect microbiome is for insect fitness, expansion into novel ecological niches, and novel environments. These investigations have garnered heightened interest recently, yet a comprehensive understanding of how intraspecific variation in the assembly and function of these insect-associated microbial communities can shape the plasticity of insects is still lacking. Most research focuses on the core microbiome associated with a species of interest and ignores intraspecific variation. We argue that microbiome variation among insects can be an important driver of evolution, and we provide examples showing how such variation can influence fitness and health of insects, insect invasions, their persistence in new environments, and their responses to global environmental changes. A and B are two stages of an individual or a population of the same species. The drivers lead to a shift in the insect associated microbial community, which has consequences for the host. The complex interplay of those consequences affects insect adaptation and evolution and influences insect population resilience or invasion.

Mating compatibility and offspring traits evaluation among different strains of Tenebrio molitor.

The fast-growing sector of insects for food and feed stimulates researchers and the industry to explore uncharted territories, such as insect breeding, to improve economically important insect fitness traits. The yellow mealworm, Tenebrio molitor L., is one of the most thoroughly studied insect species as food and feed. However, data on mating compatibility and the effect of cross-breeding between different strains on the performance and fitness of the hybrids are scarce. In the present study, we comparatively evaluated the mating compatibility between two T. molitor inbred strains (Greek and Italian) and their outbred strains, i.e., Italian (♀)-Greek (♂) and Greek (♀)-Italian (♂), as well as the performance of their hybrid offspring. Based on the results, there is good mating compatibility among adults of the strains tested. Offspring performance, quantified as larval survival and final larval weight, followed a similar pattern among the crossings examined. Even though differences were insignificant, the outbreeding of females of the Italian strain resulted in a higher cumulative number of eggs and hatching rate and higher offspring weight. The present study sheds light on the mating compatibility of different T. molitor strains and their hybrids' economically important life table characteristics to take the mass production of insects one step further.

Thiamethoxam soil contaminations reduce fertility of soil-dwelling beetles, Aethina tumida

There in increasing evidence for recent global insect declines. This is of major concern as insects play a critical role in ecosystem functionality and human food security. Even though environmental pollutants are known to reduce insect fertility, their potential effects on insect fitness remain poorly understood - especially for soil-dwelling species. Here, we show that fertility of soil-dwelling beetles, Aethina tumida, is reduced, on average, by half due to field-realistic neonicotinoid soil contaminations. In the laboratory, pupating beetles were exposed via soil to concentrations of the neonicotinoid thiamethoxam that reflect global pollution of agricultural and natural habitats. Emerged adult phenotypes and reproduction were measured, and even the lowest concentration reported from natural habitats reduced subsequent reproduction by 50%. The data are most likely a conservative estimate as the beetles were only exposed during pupation. Since the tested concentrations reflect ubiquitous soil pollution, the data reveal a plausible mechanism for ongoing insect declines. An immediate reduction in environmental pollutants is urgently required if our aim is to mitigate the prevailing loss of species biodiversity.

Macronutrient regulation in adult spotted‐wing Drosophila and nutritional impacts on susceptibility to three commonly‐used insecticides

Abstract Nutrition provides the resources necessary to carry out all the physiological functions organisms require. This has important implications for insect pest management, as polyphagous species can experience considerable nutritional variability, which may impact their response to management tactics. Many studies have shown that dietary protein and carbohydrates play a primary role in insect fitness and that insects regulate their intake of these macronutrients to reach an optimal balance or intake target. Importantly, this intake target can vary across developmental stages, gender, and environmental conditions as physiological requirements change. In this study, we determined the intake target for adult spotted‐wing Drosophila, Drosophila suzukii, an invasive polyphagous fruit pest. We found that adult D. suzukii regulate their intake to meet a 1:4–1:5 protein‐to‐carbohydrate ratio, which is slightly more carbohydrate‐biased than the larval intake target. We also examined the effect of nutritional state on susceptibility to three commonly‐used insecticides by rearing flies on diets with different field‐relevant protein‐to‐carbohydrate ratios and exposing adults to an LC50 dose of zeta‐cypermethrin, spinetoram, or pyrethrin. Overall, female flies had higher survivorship than males, and diet effects varied across insecticides. Diet had no impact in the spinetoram treatments, while the extreme carbohydrate‐ and protein‐biased diets produced the lowest survivorship and reproductive performance in the zeta‐cypermethrin treatments. Survivorship increased with dietary protein in the pyrethrin treatments but reproductive performance did not differ across diets. Our results suggest that tactics, which reduce fly access to dietary protein sources, such as sterilisation, could increase fly susceptibility to insecticides.

Cadmium exposure through the food chain reduces the parasitic fitness of Chouioia cunea to Hyphantria cunea pupae: An ecotoxicological risk to pest control

The toxicity of heavy metals on various trophic levels along the food chain has been extensively investigated, but no studies have focused on parasitic natural enemy insects. Herein, we constructed a food chain consisting of soil–Fraxinus mandshurica seedlings–Hyphantria cunea pupae–Chouioia cunea to analyze the effects of Cd exposure through food chain on the fitness of parasitic natural enemy insects and its corresponding mechanism. The results showed that the transfer of Cd between F. mandshurica leaves and H. cunea pupae and between H. cunea pupae and C. cunea was a bio-minimization effect. After parasitizing Cd-accumulated pupa, the number of offspring larvae, and the number, individual size (body weight, body length, abdomen length) and life span of offspring adults decreased significantly, while the duration of embryo development extended significantly. The contents of malondialdehyde and H2O2 in Cd-exposed offspring wasps increased significantly, accompanied by a significantly decrease in antioxidant capacity. The cellular immunity parameters significantly decreased in Cd-accumulated pupae, including the number of hemocytes, melanization activity and the expression level of cellular immunity genes (e.g. Hemolin-1 and PPO1). The humoral immunity disorder was found in the Cd-accumulated pupae, as evidenced by that the expression level of immune recognition gene (PGRP-SA), signal transduction genes (IMD, Dorsal, and Tube), as well as all antimicrobial peptide genes (e.g. Lysozym and Attacin) decreased significantly. Cd exposure decreased the content of glucose, trehalose, amino acid, and free fatty acid in H. cunea pupae. The expression of Hk2 in glycolysis pathway and the expression of Idh2, Idh3, Cs, and OGDH in TCA cycle pathway were significantly down-regulated in Cd-accumulated pupae. Taken together, exposure to Cd through the food chain causes oxidative damage on the offspring wasps and disrupts energy metabolism of the host insect, ultimately reducing the parasitic fitness of C. cunea to H. cunea pupae.

Entomopathogens infecting brown marmorated stink bugs before, during, and after overwintering.

The microsporidian, Nosema maddoxi Becnel, Solter, Hajek, Huang, Sanscrainte & Estep, infects brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), populations in North America and Asia and causes decreased fitness in infected insects. This host overwinters as adults, often in aggregations in sheltered locations, and variable levels of mortality occur over the winter. We investigated pathogen prevalence in H. halys adults before, during, and after overwintering. Population level studies resulted in detection of N. maddoxi in H. halys in 6 new US states, but no difference in levels of infection by N. maddoxi in autumn versus the following spring. Halyomorpha halys that self-aggregated for overwintering in shelters deployed in the field were maintained under simulated winter conditions (4°C) for 5 months during the 2021-2022 winter and early spring, resulting in 34.6 ± 4.8% mortality. Over the 2020-2021 and 2021-2022 winters, 13.4 ± 3.5% of surviving H. halys in shelters were infected with N. maddoxi, while N. maddoxi infections were found in 33.4 ± 10.8% of moribund and dead H. halys that accumulated in shelters. A second pathogen, Colletotrichum fioriniae Marcelino & Gouli, not previously reported from H. halys, was found among 46.7 ± 7.8% of the H. halys that died while overwintering, but levels of infection decreased after overwintering. These 2 pathogens occurred as co-infections in 11.1 ± 5.9% of the fungal-infected insects that died while overwintering. Increasing levels of N. maddoxi infection caused epizootics among H. halys reared in greenhouse cages after overwintering.

Bacterial Communities Are Less Diverse in a Strepsipteran Endoparasitoid than in Its Fruit Fly Hosts and Dominated by Wolbachia

Microbiomes play vital roles in insect fitness and health and can be influenced by interactions between insects and their parasites. Many studies investigate the microbiome of free-living insects, whereas microbiomes of endoparasitoids and their interactions with parasitised insects are less explored. Due to their development in the constrained environment within a host, endoparasitoids are expected to have less diverse yet distinct microbiomes. We used high-throughput 16S rRNA gene amplicon sequencing to characterise the bacterial communities of Dipterophagus daci (Strepsiptera) and seven of its tephritid fruit fly host species. Bacterial communities of D. daci were less diverse and contained fewer taxa relative to the bacterial communities of the tephritid hosts. The strepsipteran’s microbiome was dominated by Pseudomonadota (formerly Proteobacteria) (> 96%), mainly attributed to the presence of Wolbachia, with few other bacterial community members, indicative of an overall less diverse microbiome in D. daci. In contrast, a dominance of Wolbachia was not found in flies parasitised by early stages of D. daci nor unparasitised flies. Yet, early stages of D. daci parasitisation resulted in structural changes in the bacterial communities of parasitised flies. Furthermore, parasitisation with early stages of D. daci with Wolbachia was associated with a change in the relative abundance of some bacterial taxa relative to parasitisation with early stages of D. daci lacking Wolbachia. Our study is a first comprehensive characterisation of bacterial communities in a Strepsiptera species together with the more diverse bacterial communities of its hosts and reveals effects of concealed stages of parasitisation on host bacterial communities.

Parasite preferences for large host body size can drive overdispersion in a fly-mite association

Body size generally correlates intraspecifically with insect fitness but can also correlate with parasite abundance (number of parasites). Host preferences by parasites, and variation in host immunity, could contribute to this trend. We investigated the effect of host size on mite-fly interactions (Macrocheles subbadius and Drosophila nigrospiracula). Mites strongly preferred to infect larger flies in pair-wise choices, and larger flies were more likely to be infected and acquired more mites in infection microcosms. Preferences of parasites resulted in size-biased infection outcomes. We discuss the implications of this heterogeneity in infection on parasite overdispersion and fly populations.