Supercooling points of freeze-avoiding bumble bees vary with caste and queen life stage.

Bumble bees thrive in cold climates including high latitude and high altitude regions around the world, yet cold tolerance strategies are largely unknown for most species. To determine bumble bee cold tolerance strategy, we exposed bumble bees to a range of low temperatures and measured survival 72 h post-exposure. All bees that froze died within 72 h while only one bee died without freezing, suggesting that bumble bees are generally freeze-avoiding insects and may be slightly chill susceptible. We then assessed whether temperatures that cause internal ice formation (supercooling points, SCP) varied among bumble bee castes (drones, workers, and queens), or across queen life stages, collection elevation, species, or season. Males froze at significantly lower temperatures than workers or queens. Queens in pre-overwintering or overwintering states froze at significantly lower temperatures than queens stimulated to initiate ovary development by CO2 narcosis (i.e., "spring" queens). We also tested whether the presence of water (i.e., wet or dry) or ramping rate affected SCP. As expected, queens inoculated with water froze at significantly higher temperatures than dry queens. SCP tended to be lower, but not significantly so, at faster ramping rates (0.5 °C/min vs 0.25 °C/min). We also found no differences in SCP between queen bumble bees collected in spring and fall, between queens collected at two sites differing in elevation by 1100 m, or between three field-caught bumble bee species. Bumble bees appear to have relatively high, invariable SCPs, likely making them highly susceptible to freezing across all seasons. As bumble bees are not freeze-tolerant and appear to lack the ability to prevent freezing at temperatures much below 0 °C, they may rely on season- and caste-specific micro-habitat selection to thrive in cold climates.

Configuration of the thermal landscape determines thermoregulatory performance of ectotherms.

Although most organisms thermoregulate behaviorally, biologists still cannot easily predict whether mobile animals will thermoregulate in natural environments. Current models fail because they ignore how the spatial distribution of thermal resources constrains thermoregulatory performance over space and time. To overcome this limitation, we modeled the spatially explicit movements of animals constrained by access to thermal resources. Our models predict that ectotherms thermoregulate more accurately when thermal resources are dispersed throughout space than when these resources are clumped. This prediction was supported by thermoregulatory behaviors of lizards in outdoor arenas with known distributions of environmental temperatures. Further, simulations showed how the spatial structure of the landscape qualitatively affects responses of animals to climate. Biologists will need spatially explicit models to predict impacts of climate change on local scales.

The upper thermal tolerance of the secondary screwworm, Cochliomyia macellaria Fabricius (Diptera: Calliphoridae).

Determining the thermal tolerance of an organism is important when assessing its activity time and survival rate in a given environment. However, thermal tolerance is not a static trait and may be influenced by a number of environmental and organismal factors. We report the results of three experiments investigating the effects of environmental temperature, exposure duration, age, sex, and nutrient availability on the upper thermal tolerance of the adult secondary screwworm, Cochliomyia macellaria. The probability of knockdown and survival was determined using a static method for different environmental temperatures (22, 40, 42, 44, or 45 °C), exposure durations (1, 2, 4, or 6 h), and nutrient availabilities (no food or water, water only, or both food and water) for both sexes and two age classes (young = 7-9 days post pupal emergence, old = 10-12 days post pupal emergence). In general, environmental temperature and exposure duration had the greatest effects on both the probability of knockdown and survival. As temperature or duration increased, the probability of knockdown increased while the probability of survival decreased. The availability of nutrients (water only or food and water) increased thermal tolerance at moderate temperatures (42 and 44 °C), but had no effect at 45 °C. Female flies were more thermally tolerant than males, regardless of nutrient availability. And age exhibited negligible effects on the probabilities of knockdown or survival, regardless of nutrient availability. These data show multiple environmental factors affected the thermal tolerance of C. macellaria. Thus, such aspects of basic thermal biology should feature more prominently in applied fields using blow flies, including but not limited to forensic entomology, disease ecology, and pollination ecology.

Lizards perceived abiotic and biotic stressors independently when competing for shade in terrestrial mesocosms.

Hormones such as glucocorticoids and androgens enable animals to respond adaptively to environmental stressors. For this reason, circulating glucocorticoids became a popular biomarker for estimating the quality of an environment, and circulating androgens are frequently used to indicate social dominance. Here, we show that access to thermal resources influence the hormones and behavior of male lizards (Sceloporus jarrovi). We exposed isolated and paired males to different thermal landscapes, ranging from one large patch of shade to sixteen smaller patches. Both the presence of a competitor and the patchiness of the thermal environment influenced hormone concentrations and movement patterns. When shade was concentrated in space, paired lizards competed more aggressively and circulated more corticosterone. Even without competitors, lizards circulated more corticosterone in landscapes with fewer patches of shade. Conversely, shifts in circulating testosterone depended only on the relative body size of a lizard; when paired, large males and small males circulated more and less testosterone, respectively. Furthermore, isolated males moved the farthest and covered the most area when shade was concentrated in a single patch, but paired males did the opposite. Because the total area of shade in each landscape was the same, these hormonal and behavioral responses of lizards reflect the ability to access shade. Thus, circulating glucocorticoids should reflect the thermal quality of an environment when researchers have controlled for other factors. Moreover, a theory of stress during thermoregulation would help ecologists anticipate physiological and behavioral responses to changing climates.

Diminishing returns limit energetic costs of climate change.

Changes in the time available for organisms to maintain physiologically preferred temperatures (thermal opportunity) is a primary mechanism by which climate change impacts the fitness and population dynamics of organisms. Yet, it is unclear whether losses or gains in thermal opportunity result in proportional changes in rates of energy procurement and use. We experimentally quantified lizard food consumption and energy assimilation at different durations of thermal opportunity. We incorporated these data in an individual-based model of foraging and digestion in lizards to explore the implications of nonlinear responses to shifts in thermal opportunity across a wide geographic range. Our model predicts that shifts in thermal opportunities resulting from climate change alter energy intake primarily through digestion rather than feeding, because simulated lizards were able to fill their gut faster than they can digest their food. Moreover, since rates of energy assimilation decelerate with increasing thermal opportunity, shifts in daily energetic assimilation would depend on the previous opportunity for thermoregulation. In particular, the same changes in thermal opportunity will have little impact on lizards from warm locations, while having a large impact on lizards from cold locations where thermoregulation is possible for only a few hours each day. Energy expenditure followed spatial patterns in thermal opportunity, with greater annual energy expenditure occurring at warmer locations. Our model predicts that lizards will spend more energy under climate change by maintaining higher body temperatures and remaining active longer. However, the predicted changes in energy assimilation following climate change greatly exceeded the predicted increases in energy expenditure. Simple models, which assume constant rates of energy gain during activity, will potentially mislead efforts to understand and predict the biological impacts of climate change.

Methods and pitfalls of measuring thermal preference and tolerance in lizards.

Understanding methodological and biological sources of bias during the measurement of thermal parameters is essential for the advancement of thermal biology. For more than a century, studies on lizards have deepened our understanding of thermal ecophysiology, employing multiple methods to measure thermal preferences and tolerances. We reviewed 129 articles concerned with measuring preferred body temperature (PBT), voluntary thermal tolerance, and critical temperatures of lizards to offer: a) an overview of the methods used to measure and report these parameters, b) a summary of the methodological and biological factors affecting thermal preference and tolerance, c) recommendations to avoid identified pitfalls, and d) directions for continued progress in our application and understanding of these thermal parameters. We emphasize the need for more methodological and comparative studies. Lastly, we urge researchers to provide more detailed methodological descriptions and suggest ways to make their raw data more informative to increase the utility of thermal biology studies.

Larval diet impacts black soldier fly (Diptera: Stratiomyidae) thermal tolerance and preference.

Thermal tolerance and preference are key parameters impacting agricultural production systems. In this study, the impact of larval diet on black soldier fly thermal tolerance and preference across life-stages and sexes was examined. Larvae were fed either a low-protein high-carbohydrate synthetic diet (i.e., P7C35), a high-protein low-carbohydrate synthetic diet (i.e., P35C7), or the Gainesville diet (i.e., C) as a control and reference. Our results demonstrate that the impacts of larval diet on black soldier fly thermal tolerance and preference could be stage and sex specific. The mean heat knockdown temperatures (HKT) ranged between 46.6 and 47.9 °C. Synthetic diets resulted in greater HKT and the difference decreased form larvae (e.g., ∼1 °C) to adults (e.g., ∼0.2 °C). The mean chill-coma recovery time (CCRT) ranged between 8.3 and 21.6 min. Not much differences were detected between diets, but CCRT became longer from larvae to adults. The mean thermal preference ranged between 13.6 and 29.5 °C. Larvae fed synthetic diets preferred much lower temperatures than the control diet. A bimodal distribution was observed for adults regardless of sex. Differences on body mass, lipid, and protein contents were detected among diets; however, more research should be done before any conclusions can be linked to their thermal traits. These findings highlight the importance of considering the ingredients and nutritional makeup of larval diets when optimizing temperature management protocols for mass production of black soldier flies. Conversely, specific diets can be developed to promote survival under extreme rearing temperatures.

Black soldier fly (Diptera: Stratiomyidae) larval heat generation and management.

Mass production of black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), larvae results in massive heat generation, which impacts facility management, waste conversion, and larval production. We tested daily substrate temperatures with different population densities (i.e., 0, 500, 1000, 5000, and 10 000 larvae/pan), different population sizes (i.e., 166, 1000, and 10 000 larvae at a fixed feed ratio) and air temperatures (i.e., 20 and 30 °C) on various production parameters. Impacts of shifting larvae from 30 to 20 °C on either day 9 or 11 were also determined. Larval activity increased substrate temperatures significantly (i.e., at least 10 °C above air temperatures). Low air temperature favored growth with the higher population sizes while high temperature favored growth with low population sizes. The greatest average individual larval weights (e.g., 0.126 and 0.124 g) and feed conversion ratios (e.g., 1.92 and 2.08 g/g) were recorded for either 10 000 larvae reared at 20 °C or 100 larvae reared at 30 °C. Shifting temperatures from high (30 °C) to low (20 °C) in between (∼10-d-old larvae) impacted larval production weights (16% increases) and feed conversion ratios (increased 14%). Facilities should consider the impact of larval density, population size, and air temperature during black soldier fly mass production as these factors impact overall larval production.

High Temperature Limits of Survival and Oviposition of Phormia regina (Meigen) and Lucilia sericata (Meigen).

The temperature dependent development rates of blow flies allow blow flies to be used as biological clocks in forensic death investigations. However, the upper thermal limits of adult survival and oviposition, both required for producing larvae, remains largely unknown. Therefore, in this study we examined the impact of a range of temperatures between 37 °C and 44 °C on the likelihood of survival and egg-laying behavior of two species of medicolegal forensic importance, Lucilia sericata (Meigen) and Phormia regina (Meigen) (Diptera: Calliphoridae). To quantify the upper temperature limits of survival, adult fly colonies were exposed to 37 °C, 41 °C, 42 °C, 43 °C, and 44 °C for 24 h. Similarly for oviposition trials, adults of both species were exposed to 40 °C, 42 °C, and 43 °C with P. regina oviposition also observed at 41 °C. Trials lasted for 24 h with oviposition substrate replenished at the 12 h mark. A yes/no determination on egg deposition was made, eggs were counted, and a yes/no determination was made on egg hatch. Survival did not differ by species (p = 0.096). Overall, survival decreased with increasing temperatures, with ~100% at 37 °C, ~50% at 41 °C, ~37% at 42 °C, ~15% at 43 °C and 0% at 44 °C. Lucilia sericata laid eggs capable of hatch up to 43 °C, while Phormia regina egg-hatch was observed up to 41 °C. These results indicate a greater thermal tolerance of adult survival than for egg deposition and successful egg hatch, which supports previous experiments indicating blow flies stop laying eggs at sub-lethal temperatures. Furthermore, these data indicate that adult blow flies may find remains at or near time of death but may delay egg deposition until temperatures drop below an acceptable threshold.

The devil is in the details: Variable impacts of season, BMI, sampling site temperature, and presence of insects on the post-mortem microbiome.

Background: Post-mortem microbial communities are increasingly investigated as proxy evidence for a variety of factors of interest in forensic science. The reported predictive power of the microbial community to determine aspects of the individual's post-mortem history (e.g., the post-mortem interval) varies substantially among published research. This observed variation is partially driven by the local environment or the individual themselves. In the current study, we investigated the impact of BMI, sex, insect activity, season, repeat sampling, decomposition time, and temperature on the microbial community sampled from donated human remains in San Marcos, TX using a high-throughput gene-fragment metabarcoding approach. Materials and methods: In the current study, we investigated the impact of BMI, sex, insect activity, season, repeat sampling, decomposition time, and temperature on the microbial community sampled from donated human remains in San Marcos, TX using a high-throughput gene-fragment metabarcoding approach. Results: We found that season, temperature at the sampling site, BMI, and sex had a significant effect on the post-mortem microbiome, the presence of insects has a homogenizing influence on the total bacterial community, and that community consistency from repeat sampling decreases as the decomposition process progresses. Moreover, we demonstrate the importance of temperature at the site of sampling on the abundance of important diagnostic taxa. Conclusion: The results of this study suggest that while the bacterial community or specific bacterial species may prove to be useful for forensic applications, a clearer understanding of the mechanisms underpinning microbial decomposition will greatly increase the utility of microbial evidence in forensic casework.

Response of Culex pipiens pallens to Visual and Olfactory Stimuli from a Mosquito Trap.

This study examined Culex pipiens pallens responses to different combinations of colors and chemicals employed via a mosquito trap under semifield conditions. Our results indicated that Cx. p. pallens has color and chemical concentration preferences. Culex p. pallens had a 38.0% greater response to white than black color treated traps. Further, Cx. p. pallens showed differences in olfactory attraction depending on the chemical and concentration. Culex p. pallens was 107.6% more attracted to traps employing 500 ppm ammonia than control (i.e., unscented). Similarly, Cx. p. pallens was 117.5%, 128.8%, and 140.3% more attracted to traps employing, respectively, 1,000, 10,000, and 20,000 ppm of ammonia hydrogen carbonate compared to controls. And the response to lactic acid showed that Cx. p. pallens was most attracted to concentrations of 100 and 500 ppm (135.7% and 142.9%, respectively) compared to controls.

The World Still Is Not Flat: Lessons Learned from Organismal Interactions with Environmental Heterogeneity in Terrestrial Environments.

Over the past decade, ecologists and physiologists alike have acknowledged the importance of environmental heterogeneity. Meaningful predictions of the responses of organisms to climate will require an explicit understanding of how organismal behavior and physiology are affected by such heterogeneity. Furthermore, the responses of organisms themselves are quite heterogeneous: physiology and behavior vary over different time scales and across different life stages, and because physiological systems do not operate in isolation of one another, they need to be considered in a more integrated fashion. Here, we review case studies from our laboratories to highlight progress that has been made along these fronts and generalizations that might be made to other systems, particularly in the context of predicting responses to climate change.