Beneficial developmental acclimation in reproductive performance under cold but not heat stress

Thermal plasticity can help organisms coping with climate change. In this study, we analyse how laboratory populations of the ectotherm species Drosophila subobscura, originally from two distinct latitudes and evolving for several generations in a stable thermal environment (18 °C), respond plastically to new thermal challenges. We measured adult performance (fecundity traits as a fitness proxy) of the experimental populations when exposed to five thermal regimes, three with the same temperature during development and adulthood (15-15 °C, 18-18 °C, 25-25 °C), and two where flies developed at 18 °C and were exposed, during adulthood, to either 15 °C or 25 °C. Here, we test whether (1) flies undergo stress at the two more extreme temperatures; (2) development at a given temperature enhances adult performance at such temperature (i.e. acclimation), and (3) populations with different biogeographical history show plasticity differences. Our findings show (1) an optimal performance at 18 °C only if flies were subjected to the same temperature as juveniles and adults; (2) the occurrence of developmental acclimation at lower temperatures; (3) detrimental effects of higher developmental temperature on adult performance; and (4) a minor impact of historical background on thermal response. Our study indicates that thermal plasticity during development may have a limited role in helping adults cope with warmer - though not colder - temperatures, with a potential negative impact on population persistence under climate change. It also emphasizes the importance of analysing the impact of temperature on all stages of the life cycle to better characterize the thermal limits.     

Plastic and Genetic Variation in Wing Loading as a Function of Temperature Within and Among Parallel Clines in Drosophila subobscura

Drosophila subobscura is a European (EU) species that was introducedinto South America (SA) approximately 25 years ago. Previousstudies have found rapid clinal evolution in wing size and inchromosome inversion frequency in the SA colonists, and theseclines parallel those found among the ancestral EU populations.Here we examine thermoplastic changes in wing length in fliesreared at 15, 20, and 25°C from 10 populations on each continent.Wings are plastically largest in flies reared at 15°C (thecoldest temperature) and genetically largest from populationsthat experience cooler temperatures on both continents. We hypothesizethat flies living in cold temperatures benefit from reducedwing loading: ectotherms with cold muscles generate less powerper wing beat, and hence larger wings and/or a smaller masswould facilitate fight. We develop a simple null model, basedon isometric growth, to test our hypothesis. We find that bothEU and SA flies exhibit adaptive plasticity in wing loading:flies reared at 15°C generally have lower wing loadingsthan do flies reared at 20°C or 25°C. Clinal patterns,however, are strikingly different. The ancestral EU populationsshow adaptive clinal variation at rearing a temperature of 15°C:flies from cool climates have lower wing loadings. In the colonizingpopulations from SA, however, we cannot reject the null model:wing loading increases with decreasing clinal temperatures.Our data suggest that selective factors other than flight havefavored the rapid evolution of large overall size at low environmentaltemperatures. However, selection for increased flight abilityin such environments may secondarily favor reduced body mass.     

Effect of temperature on the development of the West Indian sweet potato weevil, <Emphasis Type="Italic">Euscepes postfasciatus</Emphasis> (Fairmaire) (Coleoptera: Curculionidae) on an artificial diet

The relationship between temperature and the development of the West Indian sweet potato weevil, Euscepes postfasciatus, on an artificial larval diet containing powdered sweet potato root, was examined at different fixed temperatures from 22 to 31°C. The developmental periods for egg, larvae, and pupae stages shortened in correlation with increased temperature. The thermal constant was 769.2 degree-days and the developmental zero for female and male was 11.1 and 11.7°C, respectively. Although we can rear this weevil at temperatures ranging from 22 to 31°C, rearing temperatures should be kept between 25 and 28°C because the developmental stages were too long at 22°C and the larval period was delayed at 31°C. The basis for these developmental data will be a useful key factor in designing a plan to eradicate the weevil by using a mass-rearing system and SIT.     

Temperature‐dependent development of the double‐spined spruce bark beetle Ips duplicatus (Sahlberg, 1836) (Coleoptera; Curculionidae)

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Development of Sarcophaga dux (diptera: Sarcophagidae) at constant temperatures and differential gene expression for age estimation of the pupae

Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly species with potential forensic value for estimating minimum postmortem interval (PMI_min). The basic developmental data and precise age estimates of the pupae are significant for PMI_min estimation in forensic investigations. In the present study, we investigated the development data of that species at seven constant temperatures varying from 16 °C to 34 °C, including body length changes of the larve, developmental duration and accumulated degree hours of the preadults. Several reference genes for relative quantification of the differentially expressed genes (DEGs) were firstly selected and evaluated in the pupae of different ages under different temperatures. The DEGs of the insects during the pupal period at different constant temperatures (34, 25 and 16 °C) were further analyzed for more precise age estimation. The results showed that the developmental durations of the preadults at 16, 19, 22, 25, 28, 31 and 34 °C were 1478.6 ± 18.3 h, 726.1 ± 15.8 h, 538.5 ± 0.9 h, 394.1 ± 9.5 h, 375.6 ± 10.8 h, 284.1 ± 7.3 h, and 252.5 ± 6.1 h, respectively. The developmental threshold temperature the flies was 12.27 ± 0.35 °C, and the thermal summation constant was 5341.71 ± 249.29° hours. The most reliable reference genes during the pupal period at different temperatures were found: GST1 and 18S rRNA for the 34 °C group, GST1 and RPL49 for 25 °C, and 18S rRNA and 28S rRNA for 16 °C. The four differential expression genes (Hsp60, A-alpha, ARP, and RPL8) have the potential to be used for more precise age estimation of pupal S. dux. This work provides important basic developmental data and a more precise age estimation method for pupal S. dux, and improves the value of this species for PMI_min estimation in forensic investigations.     

Adult plasticity of cold tolerance in a continental-temperate population of Drosophila suzukii

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a worldwide emerging pest of soft fruits, but its cold tolerance has not been thoroughly explored. We determined the cold tolerance strategy, low temperature thermal limits, and plasticity of cold tolerance in both male and female adult D. suzukii. We reared flies under common conditions (long days, 21 °C; control) and induced plasticity by rapid cold-hardening (RCH, 1 h at 0 °C followed by 1 h recovery), cold acclimation (CA, 5 days at 6 °C) or acclimation under fluctuating temperatures (FA). D. suzukii had supercooling points (SCPs) between −16 and −23 °C, and were chill-susceptible. 80% of control flies were killed after 1 h at −7.2 °C (males) or −7.5 °C (females); CA and FA improved survival of this temperature in both sexes, but RCH did not. 80% of control flies were killed after 70 h (male) or 92 h (female) at 0 °C, and FA shifted this to 112 h (males) and 165 h (females). FA flies entered chill coma (CT_min) at approximately −1.7 °C, which was ca. 0.5 °C colder than control flies; RCH and CA increased the CT_min compared to controls. Control and RCH flies exposed to 0 °C for 8 h took 30–40 min to recover movement, but this was reduced to <10 min in CA and FA. Flies placed outside in a field cage in London, Ontario, were all killed by a transient cold snap in December. We conclude that adult phenotypic plasticity is not sufficient to allow D. suzukii to overwinter in temperate habitats, and suggest that flies could overwinter in association with built structures, or that there may be additional cold tolerance imparted by developmental plasticity.     

Locomotor performance of Drosophila melanogaster: interactions among developmental and adult temperatures, age, and geography

We explored the extent to which a phenotypic trait (walking speed) of Drosophila melanogaster is influenced by population, developmental temperature, adult temperature, and age. Our goals were to estimate the importance of these factors and to test the beneficial acclimation hypothesis. We measured speed of flies from two populations (the Congo and France) that developed at different temperatures (18, 25, and 29 degrees C) and were tested at different temperatures (18, 25, and 29 degrees C) and ages (2, 7, 13 days). Not surprisingly, speed increased strongly with test temperature. Speed was generally greatest for flies reared at an intermediate developmental temperature, contrary to the beneficial acclimation hypothesis, which predicts that speed would be greatest when influenced by interactions involving population. For example, speed was greatest for flies from France that developed at a low temperature, but for flies from the Congo that developed at a high temperature. The impact of developmental temperature declined with age. Surprisingly, speed actually increased with age for flies raised and maintained at a low temperature, but decreased with age for flies raised and maintained at an intermediate or at a high temperature. Thus, walking performance is highly dynamic phenotypically, complicating potential attempts to predict responses to selection on performance.     

Strong responses of Drosophila melanogaster microbiota to developmental temperature

Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.     

Flies evolved small bodies and cells at high or fluctuating temperatures

Recent theory predicts that the sizes of cells will evolve according to fluctuations in body temperature. Smaller cells speed metabolism during periods of warming but require more energy to maintain and repair. To evaluate this theory, we studied the evolution of cell size in populations of Drosophila melanogaster held at either a constant temperature (16°C or 25°C) or fluctuating temperatures (16 and 25°C). Populations that evolved at fluctuating temperatures or a constant 25°C developed smaller thoraxes, wings, and cells than did flies exposed to a constant 16°C. The cells of flies from fluctuating environments were intermediate in size to those of flies from constant environments. Most genetic variation in cell size was independent of variation in wing size, suggesting that cell size was a target of selection. These evolutionary patterns accord with patterns of developmental plasticity documented previously. Future studies should focus on the mechanisms that underlie the selective advantage of small cells at high or fluctuating temperatures.     

Cold acclimation allows Drosophila flies to maintain mitochondrial functioning under cold stress

Environmental stress generally disturbs cellular homeostasis. Researchers have hypothesized that chilling injury is linked to a shortage of ATP. However, previous studies conducted on insects exposed to nonfreezing low temperatures presented conflicting results. In this study, we investigated the mitochondrial bioenergetics of Drosophila melanogaster flies exposed to chronic cold stress (4 °C). We assessed mitochondrial oxygen consumption while monitoring the rate of ATP synthesis at various times (0, 1, 2, and 3 days) during prolonged cold stress and at two assay temperatures (25 and 4 °C). We compared organelle responses between cold-susceptible and cold-acclimated phenotypes. Continuous exposure to low temperature provoked temporal declines in the rates of mitochondrial respiration and ATP synthesis. Respiratory control ratios (RCRs) suggested that mitochondria were not critically uncoupled. Nevertheless, after 3 days of continuous cold stress, a sharp decline in the mitochondrial ATP synthesis rate was observed in control flies when they were assayed at low temperature. This change was associated with reduced survival capacity in control flies. In contrast, cold-acclimated flies exhibited high survival and maintained higher rates of mitochondrial ATP synthesis and coupling (i.e., higher RCRs). Adaptive changes due to cold acclimation observed in the whole organism were thus manifested in isolated mitochondria. Our observations suggest that cold tolerance is linked to the ability to maintain bioenergetics capacity under cold stress.     

A model for blood meal digestion and fat metabolism in male tsetse flies (Glossinidae)

Abstract. Fat and haematin levels of mature male Glossina morsitans morsitans Westwood were estimated at different times after feeding at temperatures between 15 and 30°C. Flies were kept (largely inactive) in 7.5 × 2.5 cm tubes, or in actograph cages, where flight activity increased with time after feeding. Haematin excretion was modelled as a series of three first order reactions, all with the same rate parameter. The model accounted for > 98% of the variance in mean haematin in each of seven experiments; the rate parameter increased linearly with temperature and activity level. A similar approach was adopted for modelling fat metabolism. The rate coefficients of lipogenesis increased with temperature, and that for lipolysis with temperature, activity level and their interaction. All experiments were analysed simultaneously to provide equations predicting haematin or fat levels for all times, for active or inactive flies, and for temperatures between 15 and 30°C. Haematin exhibited large variations between individuals, but for active flies the expected haematin content at a given time varied little between flies kept at 25 and at 30°C. In inactive flies kept at 25°C, lipogenesis peaked at ≈ 24 h and lipolysis at ≈ 48 h. For active flies the times were 12 and 24 h, respectively; both rates were about twice as high as in inactive flies. Active flies produced (up to 1 mg) more fat out of a given size of blood meal than inactive flies. Curves of fat content against logarithm of haematin content differed little with temperature, and can therefore be useful for comparative studies of field populations of tsetse.     

Effect of temperature on immature development and longevity of two introduced opiine parasitoids on Bactrocera invadens

Temperature‐dependent development, parasitism and longevity of the braconid parasitoids, Fopius arisanus Sonan and Diachasmimorpha longicaudata Ashmed on Bactorcera invadens Drew Tsuruta & White, was evaluated across five constant temperatures (15, 20, 25, 30 and 35°C). Developmental rate decreased linearly with increasing temperature for both the parasitoid species. Linear and Brière‐2 nonlinear models were used to determine the lower temperature threshold at which the developmental rate (1/D) approached zero. For F. arisanus, lower thresholds to complete development estimated with the linear and nonlinear models were 10.1 and 6.9°C, respectively. The total degree‐days (DD) required to complete the development estimated by the linear model for F. arisanus was 360. In D. longicaudata, the linear and nonlinear models estimated lower thresholds of 10.4 and 7.3°C, respectively, and the total DD estimated was 282. In F. arisanus, percentage parasitism differed significantly across all temperatures tested and was highest at 25°C (71.1 ± 2.5) and lowest at 15°C (46.4 ± 1.4). Parasitoid progeny sex ratio was female biased at all temperatures except at 20°C. In D. longicaudata, percentage parasitism was highest at 20°C (52.2 ± 4.0) and lowest at 15°C (27.7 ± 2.5). Parasitoid progeny sex ratio was female biased and similar for all temperatures. Adult longevity of both parasitoids was shortest at 35°C and longest at 15°C, and females lived significantly longer than males at all temperatures tested. Our findings provide some guidance for future mass rearing and field releases of the two parasitoids for the management of B. invadens in Africa.     

Temperature-dependent development of Lista haraldusalis (Walker) (Lepidoptera: Pyralidae) on Platycarya strobilacea

The effect of constant temperatures on development and survival of Lista haraldusalis (Walker) (Lepidoptera: Pyralidae), a newly reported insect species used to produce insect tea in Guizhou province (China), was studied in laboratory conditions at seven temperatures (19 °C, 22 °C, 25 °C, 28 °C, 31 °C, 34 °C, and 37 °C) on Platycarya strobilacea. Increasing the temperature from 19 °C to 31 °C led to a significant decrease in the developmental time from egg to adult emergence, and then the total developmental time increased at 34 °C. Egg incubation was the stage where L. haraldusalis experienced the highest mortality at all temperatures. The survival of L. haraldusalis was significantly higher at 25 °C and 28 °C, whereas none of the eggs hatched at 37 °C. Common and Ikemoto linear models were used to describe the relationship between the temperature and the developmental rate for each immature stage of L. haraldusalis. The estimated values of the lower temperature threshold and thermal constant of the total immature stages using Common and Ikemoto linear models were 11.34 °C and 11.20 °C, and 939.85 and 950.41 degree-days, respectively. Seven nonlinear models were used to fit the experimental data to estimate the developmental rate of L. haraldusalis. Based on the biological significance for model evaluation, Ikemoto linear, Logan-6, and SSI were the best models that fitted each immature stage of L. haraldusalis and they were used to estimate the temperature thresholds. These thermal requirements and temperature thresholds are crucial for facilitating the development of factory-based mass rearing of L. haraldusalis.     

The combined effects of temperature and diet on development and survival of a crab spider,Misumenops tricuspidatus (Fabricius) (Araneae: Thomisidae)

In the present study, the combined effects of temperature and diet on development and survival of a crab spider, Misumenops tricuspidatus (Fabricius) (Araneae: Thomisidae) in laboratory conditions were investigated. The experiments were carried out at five constant temperatures ranging from 15°C to 35°C on two kinds of diets, fruit flies (Drosophila melanogaster) and a mixed diet of fruit flies and dung flies. It was found that development rate of eggs increased with successive temperature increments, reaching a maximum at 30°C, then declined at 32°C and that eggs survived well between 20°C and 30°C (>70%), but no eggs survived to hatching at 35°C regardless of whether the spiders were fed on single or mixed diet. Juveniles completed development on both diets at all constant temperatures tested, but survival was low at the extreme temperatures. Juvenile development times decreased over successive temperature increments up to 30°C, then increased at 32°C. Females developed faster than males. Diet also influenced development time, survival and number of moults to reach maturity. Juveniles raised on the mixed diet composed of fruit flies and dung flies developed faster, survived better, and required fewer moults to reach maturity than on a diet composed of only fruit flies. Plots of development rates (reciprocal of mean development times) and survival rates (expressed as percentages) against constant temperatures indicated that M. tricuspidatus is well adapted to low temperatures, but detrimentally affected by high temperatures. Using linear regression, the lower development threshold (LDT) and the sum of effective temperatures (SET) for all life stages of M. tricuspidatus on each diet were estimated. LDT and SET varied among developmental stages and between diets.     

Temperature-dependent physiological and biochemical responses of the marine medaka Oryzias melastigma with consideration of both low and high thermal extremes

This study aimed to investigate temperature effect on physiological and biochemical responses of the marine medaka Oryzias melastigma larvae. The fish were subjected to a stepwise temperature change at a rate of 1 °C/h increasing or decreasing from 25 °C (the control) to six target temperatures (12, 13, 15, 20, 28 and 32 °C) respectively, followed by a 7-day thermal acclimation at each target temperature. The fish were fed ad libitum during the experiment. The results showed that cumulative mortalities were significantly increased at low temperatures (12 and 13 °C) and at the highest temperature (32 °C). For the survivors, their growth profile closely followed the left-skewed ‘thermal performance curve’. Routine oxygen consumption rates of fish larvae were significantly elevated at 32 °C but suppressed at 13 and 15 °C (due to a high mortality, larvae from 12 °C were not examined). Levels of heat shock proteins and activities of malate dehydrogenase and lactate dehydrogenase were also measured in fish larvae exposed at 15, 25 and 32 °C. The activities of both enzymes were significantly increased at both 15 and 32 °C, where the fish larvae probably suffered from thermal discomfort and increased anaerobic components so as to compensate the mismatch of energy demand and supply at these thermal extremes. Coincidently, heat shock proteins were also up-regulated at both 15 and 32 °C, enabling cellular protection. Moreover, the critical thermal maxima and minima of fish larvae increased significantly with increasing acclimation temperature, implying that the fish could develop some degrees of thermal tolerance through temperature acclimation.     

Effects of constant and fluctuating incubation temperatures on hatching success and hatchling traits in the diamondback terrapin (Malaclemys terrapin) in the context of the warming climate

As global temperatures continue to rise, so too will the nest temperatures of many species of turtles. Yet for most turtle species, including the estuarine diamondback terrapin (Malaclemys terrapin), there is limited information on embryonic sensitivity to elevated temperature. We incubated eggs of M. terrapin at three, mean temperatures (31, 34, 37 °C) under two thermal exposure regimes (constant or semi-naturally fluctuating temperature) and measured hatching success, developmental rate, and hatchling size. Hatching success was 100% at 31 °C and 67% at 34 °C, respectively; at 37 °C, all eggs failed early in the incubation period. These values were unaffected by exposure regime. The modeled LT₅₀ (temperature that was lethal to 50% of the test population) was 34.0 °C in the constant and 34.2 °C in the fluctuating thermal regime, reflecting a steep decline in survival between 33 and 35 °C. Hatchlings having been incubated at a constant 34 °C hatched sooner than those incubated at 31 °C under either constant or fluctuating temperature. Hatchlings were smaller in straight carapace length (CL) and width after having been incubated at 34 °C compared to 31 °C. Larger (CL) hatchlings resulted from fluctuating temperature conditions relative to constant temperature conditions, regardless of mean temperature. Based upon recent temperatures in natural nests, the M. terrapin population studied here appears to possess resiliency to several degrees of elevated mean nest temperatures, beyond which, embryonic mortality will likely sharply increase. When considered within the mosaic of challenges that Maryland's M. terrapin face as the climate warms, including ongoing habitat losses due to sea level rise and impending thermal impacts on bioenergetics and offspring sex ratios, a future increase in embryonic mortality could be a critical factor for a population already experiencing ecological and physiological challenges due to climate change.     

Cross-mating experiments with geographically different populations of <Emphasis Type="Italic">Amblyomma cajennense</Emphasis> (Acari: Ixodidae)

The effect of temperature on the development and fecundity of Sancassania polyphyllae fed on tissues of Polyphylla fullo larvae was studied at 15, 20, 25, 30, and 35 ± 1°C and 65 ± 10% RH in a dark incubator. Mean developmental period of immature stages decreased significantly with increasing temperatures from 15 to 30°C. Developmental periods at 30–35°C were not significantly different. The estimated lower developmental thresholds of the various immature stages ranged between 10.1 and 11.5°C. The thermal constant for the egg-to-female adult was 93.5 degree-days. The pre-oviposition, oviposition, and post-oviposition periods and female longevity were significantly longer at 15°C than at higher temperatures. Mean total and daily fecundity were the highest at 25°C, which were significantly different from those obtained at 15, 20 and 30°C. The net reproductive rate (R ₀) was the highest at 25°C (588.3 ♀/♀). The longest mean generation time (T ₀) occurred at 15°C (36 days) and the shortest occurred at 30°C (9.2 days). The highest intrinsic rate of increase (r _m) for S. polyphyllae was observed at 25 (0.61 ♀/♀/day) and 30°C (0.62 ♀/♀/day).     

Root–shoot communication of the seedlings of Japanese larch and a hybrid species grown in different soil-temperature regimes

We studied the effects of soil temperature (7, 15, and 25°C) on the growth and photosynthesis of seedlings of the Japanese larch (Larix kaempferi) and its hybrid larch (L. gmelinii × L. kaempferi) to simulate early stages of regeneration after disturbance. At a soil temperature of 7°C, the root length per unit root biomass, chlorophyll concentration, and photosynthetic nitrogen-use efficiency (PNUE) were markedly lower in the Japanese larch than in the hybrid larch, which may indicate that the hybrid larch is better at acquiring water and nutrients. At ambient temperatures of 17–25°C, the light-saturated photosynthesis rate (P _sat) of both seedlings grown at a soil temperature of 7°C was lower than at 15 or 25°C. By the 16th week, the needle area, root area, and biomass in seedlings of both types were lower at a soil temperature of 7°C than at soil temperatures of 15 or 25°C. At a soil temperature of 25°C, P _sat and nitrogen uptake were lower in both larch species than at 15°C. The growth of the Japanese larch declined sharply from 15 to 25°C; however, the growth of the hybrid larch decreased only slightly from 15 to 25°C. We conclude that an increased soil temperature may retard larch growth in cold regions, especially in the case of the Japanese larch.     

Development of microsporidia-infected Muscidifurax raptor (Hymenoptera: Pteromalidae) at different temperatures

Muscididfurax raptor, a pupal parasitoid of house flies and other filth flies, is commonly infected with the microsporidium Nosema muscidifuracis. To determine the effects of infection on developmental time, uninfected and infected adult M. raptor were allowed to parasitize pupae of the house fly (Musca domestica) for 24 h. Exposed pupae of the two groups (infected and uninfected) were held at 15, 20, 25, 30, 32, and 34 °C with 75–80% relative humidity. Development of infected M. raptor was significantly longer at all temperatures than that of uninfected parasitoids, resulting in approximately 7% extensions of developmental times. Uninfected females completed development in 14.6, 19.6, and 30.4 days at 30, 25, and 20 °C, respectively, compared with 15.8, 20.7, and 32.3 days for infected females at these temperatures. The differences in developmental times provided narrow windows for isolating large proportions of uninfected M. raptor females for disease management programs. This window was greatest at 20 °C; 61% of the uninfected females emerged by day 30, at which time only 10% of the infected females had emerged.     

Explicit formula of finite difference method to estimate human peripheral tissue temperatures during exposure to severe cold stress

During cold exposure, peripheral tissues undergo vasoconstriction to minimize heat loss to preserve the maintenance of a normal core temperature. However, vasoconstricted tissues exposed to cold temperatures are susceptible to freezing and frostbite-related tissue damage. Therefore, it is imperative to establish a mathematical model for the estimation of tissue necrosis due to cold stress. To this end, an explicit formula of finite difference method has been used to obtain the solution of Pennes' bio-heat equation with appropriate boundary conditions to estimate the temperature profiles of dermal and subdermal layers when exposed to severe cold temperatures. The discrete values of nodal temperature were calculated at the interfaces of skin and subcutaneous tissues with respect to the atmospheric temperatures of 25 °C, 20 °C, 15 °C, 5 °C, −5 °C and −10 °C. The results obtained were used to identify the scenarios under which various degrees of frostbite occur on the surface of skin as well as the dermal and subdermal areas. The explicit formula of finite difference method proposed in this model provides more accurate predictions as compared to other numerical methods. This model of predicting tissue temperatures provides researchers with a more accurate prediction of peripheral tissue temperature and, hence, the susceptibility to frostbite during severe cold exposure.