A protocol for analysing thermal stress in insects using infrared thermography

The study of insect responses to thermal stress has involved a variety of protocols and methodologies that hamper the ability to compare results between studies. For that reason, the development of a protocol to standardize thermal assays is necessary. In this sense, infrared thermography solves some of the problems allowing us to take continuous temperature measurements without handling the individuals, an important fact in cold-blooded organisms like insects. Here, we present a working protocol based on infrared thermography to estimate both cold and heat thermal stress in insects. We analyse both the change in the body temperature of individuals and their behavioural response. In addition, we used partial least squares regression for the statistical analysis of our data, a technique that solves the problem of having a large number of variables and few individuals, allowing us to work with rare or endemic species. To test our protocol, we chose two species of congeneric, narrowly distributed dung beetles that are endemic to the southeastern part of the Iberian Peninsula. With our protocol we have obtained five variables in the response to cold and twelve in the response to heat. With this methodology we discriminate between the two flightless species of Jekelius through their thermal response. In response to cold, Jekelius hernandezi showed a higher rate of cooling and reached higher temperatures of stupor and haemolymph freezing than Jekelius punctatolineatus. Both species displayed similar thermoregulation ranges before reaching lethal body temperature with heat stress. Overall, we have demonstrated that infrared thermography is a suitable method to assess insect thermal responses with a high degree of sensitivity, allowing for the discrimination between closely related species.     

The effect of body fat percentage and body fat distribution on skin surface temperature with infrared thermography

This study aimed to search for relations between body fat percentage and skin temperature and to describe possible effects on skin temperature as a result of fat percentage in each anatomical site. Women (26.11±4.41 years old) (n =123) were tested for: body circumferences; skin temperatures (thermal camera); fat percentage and lean mass from trunk, upper and lower limbs; and body fat percentage (Dual-Energy X-Ray Absorptiometry). Values of minimum (T_Mi), maximum (T_Ma), and mean temperatures (T_Me) were acquired in 30 regions of interest. Pearson's correlation was estimated for body circumferences and skin temperature variables with body fat percentage. Participants were divided into groups of high and low fat percentage of each body segment, of which T_Me values were compared with Student's t-test. Linear regression models for predicting body fat percentage were tested. Body fat percentage was positively correlated with body circumferences and palm temperatures, while it was negatively correlated with most temperatures, such as T_Ma and T_Me of posterior thighs (r =−0.495 and −0.432), T_Me of posterior lower limbs (r =−0.488), T_Ma of anterior thighs (r =−0.406) and T_Mi and T_Me of posterior arms (r =−0.447 and −0.430). Higher fat percentages in the specific anatomical sites tended to decrease T_Me, especially in posterior thighs, shanks and arms. Skin temperatures and body circumferences predicted body fat percentage with 58.3% accuracy (R =0.764 and R² =0.583). This study clarifies that skin temperature distribution is influenced by the fat percentage of each body segment.     

Differences in body temperature between black-and-white and red-and-white Holstein cows reared on a hot climate using infrared thermography

Heat stress is one of the greatest issues of the dairy industry in regions with hot climate. Since coat color appears to be related to heat stress adaptiveness, we compared rectal temperatures and surface temperatures of Red-and-white (RW, n = 14) and Black-and-white (BW, n = 16) Holstein cows using infrared thermography in both cold (July; mean temperature: 15.5 °C) and hot (March; mean temperature: 30.5 °C) seasons in Southern Brazil. Thermographic images were taken from the left side of the animal at a distance of 4 m. The images obtained were then analyzed using the software Testo IRSoft. The variables obtained by thermography of the body surface include the temperature of non-pigmented patches, obtained using the average of five spots on white patches in a rectangle drawn on the body of the cow from the scapula to the ilium of the cow until the middle of the ribs; the temperature of pigmented patches, obtained using an average of 5 pigmented spots on the same rectangle; the temperature at the hottest spot and the temperature at the coldest spot, within the same rectangle. Rectal temperature measures were taken by a mercury thermometer during milkings. In our findings, during the cold season, RW cows had lower temperatures on the surface of pigmented spots (p = 0.01) but did not differ from BW animals when comparing rectal temperatures (p = 0.70). During the hot season, however, RW cows had lower temperatures on white spots (p = 0.049) as well as lower rectal temperatures (p = 0.029). These results suggest that the red coat phenotype presents less absorption of solar radiation, retaining less heat.     

Development and validity of a universal empirical equation to predict skin surface temperature on thermal manikins

Clothing evaporative resistance is an important input in thermal comfort models. Thermal manikin tests give the most accurate and reliable evaporative resistance values for clothing. The calculation methods of clothing evaporative resistance require the sweating skin surface temperature (i.e., options 1 and 2). However, prevailing calculation methods of clothing evaporative resistance (i.e., options 3 and 4) are based on the controlled nude manikin surface temperature due to the sensory measurement difficulty. In order to overcome the difficulty of attaching temperature sensors to the wet skin surface and to enhance the calculation accuracy on evaporative resistance, we conducted an intensive skin study on a thermal manikin ‘Tore’. The relationship among the nude manikin surface temperature, the total heat loss and the wet skin surface temperature in three ambient conditions was investigated. A universal empirical equation to predict the wet skin surface temperature of a sweating thermal manikin was developed and validated on the manikin dressed in six different clothing ensembles. The skin surface temperature prediction equation in an ambient temperature range between 25.0 and 34.0 °C is T_sk=34.0–0.0132HL. It is demonstrated that the universal empirical equation is a good alternative to predicting the wet skin surface temperature and facilitates calculating the evaporative resistance of permeable clothing ensembles. Further studies on the validation of the empirical equation on different thermal manikins are needed however.     

Respiratory heat loss of Holstein cows in a tropical environment

In order to develop statistical models to predict respiratory heat loss in dairy cattle using simple physiological and environmental measurements, 15 Holstein cows were observed under field conditions in a tropical environment, in which the air temperature reached up to 40°C. The measurements of latent and sensible heat loss from the respiratory tract of the animals were made by using a respiratory mask. The results showed that under air temperatures between 10 and 35°C sensible heat loss by convection decreased from 8.24 to 1.09 W m^–2, while the latent heat loss by evaporation increased from 1.03 to 56.51 W m^–2. The evaporation increased together with the air temperature in almost a linear fashion until 20°C, but it became increasingly high as the air temperature rose above 25°C. Convection was a mechanism of minor importance for respiratory heat transfer. In contrast, respiratory evaporation was an effective means of thermoregulation for Holsteins in a hot environment. Mathematical models were developed to predict both the sensible and latent heat loss from the respiratory tract in Holstein cows under field conditions, based on measurements of the ambient temperature, and other models were developed to predict respiration rate, tidal volume, mass flow rate and expired air temperature as functions of the ambient temperature and other variables.This paper forms part of A. S. Campos Maias doctoral thesis.     

Sensible and latent heat loss from the body surface of Holstein cows in a tropical environment

The general principles of the mechanisms of heat transfer are well known, but knowledge of the transition between evaporative and non-evaporative heat loss by Holstein cows in field conditions must be improved, especially for low-latitude environments. With this aim 15 Holstein cows managed in open pasture were observed in a tropical region. The latent heat loss from the body surface of the animals was measured by means of a ventilated capsule, while convective heat transfer was estimated by the theory of convection from a horizontal cylinder and by the long-wave radiation exchange based on the Stefan–Boltzmann law. When the air temperature was between 10 and 36°C the sensible heat transfer varied from 160 to –30 W m^–2, while the latent heat loss by cutaneous evaporation increased from 30 to 350 W m^–2. Heat loss by cutaneous evaporation accounted for 20–30% of the total heat loss when air temperatures ranged from 10 to 20°C. At air temperatures >30°C cutaneous evaporation becomes the main avenue of heat loss, accounting for approximately 85% of the total heat loss, while the rest is lost by respiratory evaporation.Part of first authors doctoral thesis     

Monitoring changes in skin temperature associated with exercise in horses on a water treadmill by use of infrared thermography

Infrared thermography (IRT) was used to assess surface temperature change as an indirect measure of muscle activity and exercise associated changes in blood flow in the working hind limb muscles of horses (n=7) undergoing water treadmill exercise. Three treatments were investigated including the treadmill ran dry (TD), water at the height of the proximal interphalangeal joint (PIP) and water at the height of the carpus (CP). Maximum skin surface temperature was recorded from the region of the semitendinosus muscle during exercise at each water height. There was a significant difference in surface hind limb temperature between exercise on the water treadmill ran dry and with water at the height of the PIP and CP (P<0.0001) with hotter temperatures recorded during the TD treatment. There was a greater increase in surface temperature of the hind limbs from pre exercise to maximum temperature during the PIP and CP treatments when compared to the TD treatment, however, this was not significant (P=0.58). There was no significant difference in surface hind limb temperature found between exercise in water at the height of the PIP and water at the height of the CP. The findings from this study suggest that IRT is able to non-invasively detect muscle activity and associated changes in blood flow whilst horses are exercised on a water treadmill. IRT could potentially be used as an alternative method to assess muscle activity and temperature change in an aquatic environment where existing methods present methodological challenges.     

Body temperature in premature infants during the first week of life: Exploration using infrared thermal imaging

BackgroundHypothermia is a problem for very premature infants after birth and leads to increased morbidity and mortality. Previously we found very premature infants exhibit abnormal thermal patterns, keeping foot temperatures warmer than abdominal temperatures for their first 12 h of life.PurposeWe explored the utility of infrared thermography as a non-invasive method for measuring body temperature in premature infants in an attempt to regionally examine differential temperatures.ResultsOur use of infrared imaging to measure abdominal and foot temperature for extremely premature infants in heated, humid incubators was successful and in close agreement using Bland and Altman technique with temperatures measured by skin thermistors.ConclusionsOur study methods demonstrated that it was feasible to capture full body temperatures of extremely premature infants while they were resting in a heated, humid incubator using a Flir SC640 infrared camera. This technology offers researchers and clinicians a method to examine acute changes in perfusion differentials in premature infants which may lead to morbidity.     

Comparison of abdominal skin temperature between fertile and infertile women by infrared thermography: A diagnostic approach

This retrospective study aimed to evaluate the differences in abdominal temperature (AT) between fertile (n=206; age) and infertile (n=250) women between the ages of 30 and 39 years. We evaluated the differences in two distinctive skin temperatures by thermography: ΔT₁ (CV8 index) – difference in temperature between the mid-abdomen (CV8 acupuncture area) and ventral upper arm (VUA) and ΔT₂ (CV4 index) – difference in temperature between the lower abdomen (CV4 acupuncture area) and VUA. The results indicated that the ΔT₁ and ΔT₂ of infertile women were significantly lower (by 1.05 °C and 0.79 °C, respectively; p<0.001, both) compared to those of fertile women. Additionally, the area under the curve of ΔT₁ (0.78) was greater compared to that of ΔT₂ (0.736), and its threshold was set at 0.675 °C, by which, the sensitivity and specificity of ΔT₁ for determination of fertility were found to be 80.8% and 68.4%, respectively. In conclusion, infertility is associated with lower AT. The decrease in AT in infertile women might be due to poor blood perfusion to the core muscles and tissues of the body. These findings provide a basis for further research for evaluation of clinical feasibility of thermography for analysis of infertility in women. Further evaluation of the influence of AT on fertility outcomes is required to determine the causal relationship between AT and infertility.     

The effect of fleece on core and rumen temperature in sheep

Temperature loggers were implanted to record core body temperature (T_core) and rumen temperature (T_rumen) in sheep. The relationship between T_core and T_rumen was compared for fleeced and shorn Merino sheep over a range of environmental temperatures and during stressors involved with shearing. Fleeced sheep maintained higher T_core and T_rumen than shorn sheep in all environmental conditions tested (from thermoneutral up to 33 °C and 55% relative humidity). Shearing of the fleeced sheep resulted in those sheep having a lower T_core when exposed to hot conditions, compared to the previously shorn sheep. Respiratory rates of fleeced sheep followed similar patterns and were higher than shorn sheep under all environmental conditions. After the fleeced sheep were shorn, their respiratory rates decreased to rates similar to the previously shorn sheep when under heat load, suggesting heat loss other than respiratory evaporative heat loss was augmented.     

Prediction of temperature profiles in the human skin and subcutaneous tissues

Exact mathematical solutions in terms of confluent hypergeometric and Airy's functions are obtained to study the steady state temperature distributions in human skin and subcutaneous tissues (SST). It is assumed that the skin is exposed to an air environment and heat transfer from the skin occurs by convection, radiation and evaporation. A mathematical model of the SST, accounting for heat conduction, perfusion of the capillary beds and metabolic heat productions of the dermis and subcutaneous tissues, has been solved to obtain interface temperatures for a wide range of environmental temperatures, rates of evaporation of sweat, wind speeds and relative humidities. The solutions provide inter-relationships between interface temperatures, thermal conductivities, metabolic heat production, blood perfusion, thicknesses of various layers of SST and ambient temperature.     

Reliability and validity of skin temperature measurement by telemetry thermistors and a thermal camera during exercise in the heat

New technologies afford convenient modalities for skin temperature (T_SKIN) measurement, notably involving wireless telemetry and non-contact infrared thermometry. The purpose of this study was to investigate the validity and reliability of skin temperature measurements using a telemetry thermistor system (TT) and thermal camera (TC) during exercise in a hot environment. Each system was compared against a certified thermocouple, measuring the surface temperature of a metal block in a thermostatically controlled waterbath. Fourteen recreational athletes completed two incremental running tests, separated by one week. Skin temperatures were measured simultaneously with TT and TC compared against a hard-wired thermistor system (HW) throughout rest and exercise. Post hoc calibration based on waterbath results displayed good validity for TT (mean bias [MB]=−0.18 °C, typical error [TE]=0.18 °C) and reliability (MB=−0.05 °C, TE=0.31 °C) throughout rest and exercise. Poor validity (MB=−1.4 °C, TE=0.35 °C) and reliability (MB=−0.65 °C, TE=0.52 °C) was observed for TC, suggesting it may be best suited to controlled, static situations. These findings indicate TT systems provide a convenient, valid and reliable alternative to HW, useful for measurements in the field where traditional methods may be impractical.     

Temperatures of skin, subcutaneous tissue, muscle and core in resting men in cold, comfortable and hot conditions

To examine the core-shell model of temperature distribution and the possible role of subcutaneous temperature in heat regulation, comprehensive temperature measurements were made on six nude resting men exposed for 2-3 h to comfort (27 degrees C), cold (15 degrees C) and heat (45 degrees C). Cold produced strong shivering and heat caused heavy sweating. Temperatures were recorded every 10 min from: esophagus, rectum and auditory canal; back muscle and thigh muscle at 20 mm and 40 mm depths; 6 subcutaneous sites; and 16 skin sites. Average temperatures at these 29 sites were tabulated at the ends of comfort, hot and cold and the onsets of sweating and shivering. Body temperature changes were slow to develop, the skin temperatures being fastest, and successively deeper tissues progressively slower. There was occasional after-drop and after-rise. The data were consistent with the core-shell concept. The temperature gradient from subcutaneous tissue to skin, which differed substantially with comfort, the onset of shivering and the onset of sweating, could serve as a regulatory signal. The data are now in computer format and may be of interest to biothermal modelers.     

Thermal ecophysiology of a native and an invasive gecko species in a tropical dry forest of Mexico

For ectotherms, thermal physiology plays a fundamental role in the establishment and success of invasive species in novel areas and, ultimately, in their ecological interactions with native species. Invasive species are assumed to have a greater ability to exploit the thermal environment, higher acclimation capacities, a wider thermal tolerance range, and better relative performance under a range of thermal conditions. Here we compare the thermal ecophysiology of two species that occur in sympatry in a tropical dry forest of the Pacific coast of Mexico, the microendemic species Benedetti's Leaf-toed Gecko (Phyllodactylus benedettii) and the invasive Common House Gecko (Hemidactylus frenatus). We characterized their patterns of thermoregulation, thermoregulatory efficiency, thermal tolerances, and thermal sensitivity of locomotor performance. In addition, we included morphological variables and an index of body condition to evaluate their effects on the thermal sensitivity of locomotor performance in these species. Although the two species had similar selected temperatures and thermal tolerances, they contrasted in their thermoregulatory strategies and thermal sensitivity of locomotor performance. Hemidactylus frenatus had a higher performance than the native species, P. benedettii, which would represent an ecological advantage for the former species. Nevertheless, we suggest that given the spatial and temporal limitations in habitat use of the two species, the probability of agonistic interactions between them is reduced. We recommend exploring additional biotic attributes, such as competition, behavior and niche overlap in order assess the role of alternative factors favoring the success of invasive species.     

Design and performance of a rugged standard operative temperature thermometer for avian studies

The lack of a truly satisfactory sensor which can characterize the thermal environment at the spatial scale experienced by small endotherms has hindered study of their thermoregulatory behavior. We describe a general design for a rugged, easily constructed sensor to measure standard operative temperature, T_es. We present specific designs for adult dark-eyed juncos (Junco hyemalis) and hatchling mallards (Anas platyrhynchos). Sensor response was stable and repeatable (±1.4%) over the course of several months. Over the range of conditions for which validation data were available (variable air temperature and wind with negligible net radiation), sensors predicted the mean net heat production of live animals to within ±0.023 W (equivalent to ±1°C at T_es=15°C). The main limit on accuracy was scatter in the data on metabolism and evaporative water loss in live animals. These sensors are far more rugged and easily constructed than the heated taxidermic mounts previously used to measure T_es. These characteristics facilitate the use of significant numbers of sensors in thermal mapping studies of endotherms.     

Maximum voluntary temperature of insect larvae reveals differences in their thermal biology

We investigate the thermoregulatory behaviors of larvae of four species of Drosophila (D. melanogaster, D. subobscura, D. pseudoobscura, and D. mojavensis), a thermotolerant strain of Drosophila melanogaster (T strain) known to differ in thermal biology, and two mutant stocks of D. melanogaster that have (as adults) defective thermoregulatory behavior. We describe and evaluate new techniques to measure two indices of maximum voluntary temperature of insect larvae. Both measures were highly repeatable within lines (species, strains, or mutants). One measure (temperature at which larvae stood upright) differed among lines consistent with expectations based on adult thermal ecology, suggesting that this measure will be useful measures of thermoregulatory set-points of larvae. The second measure (temperature of emergence from media) is less discriminatory.     

Modeling heat stress effects on dairy cattle milk production in a tropical environment using test-day records and random regression models

In tropical environments, dairy cattle production is constrained by several factors, including climate. The seasonal loss of milk due to heat stress is a recurring challenge for many dairy producers. The objective of this study was to detect heat stress thresholds, milk yield loss and individual animal variations using random regression models for dairy cattle from test-day milk records. Data were obtained from the Kenya Livestock Breeders Organization for the years 2000–2017 and merged with weather data. The weather parameters were grid-interpolated solar and meteorological data obtained from the National Aeronautics and Space Administration/Prediction Of Worldwide Energy Resources (NASA/POWER). After editing, the records comprised 49 993, 45 251 and 36 136 test-day records for first, second, and third lactations, respectively, for the four main dairy breeds: Friesian (68.0%), Ayrshire (21.1%), Jersey (7.6%) and Guernsey (3.3%). Variance components were estimated using Restricted Maximum Likelihood in ASReml software. Random regression models with third-order Legendre polynomials were fitted to the average and individual lactation curves and the reaction norms. An extended factor analytic variance structure for the random cow effects was used to estimate (co)variances between days in milk and thermal load. The daily average temperature (TA) and temperature humidity index (THI) were identified as the most suitable thermal load indicators for assessing milk yield losses. Considering a one day lag, the estimated heat stress thresholds were about 22 °C and 69 index units for TA and THI, respectively. Almost no differences were observed for estimated residual variances between the thermal load indicators, indicating there was no better model fit by TA or THI. The heat stress thresholds and milk loss patterns are important for management of dairy production systems in the tropics with climatic conditions similar to this study. Data recording should be improved as a tool to monitor the expected impacts of climate change and mitigation measures.     

Effects of heat acclimatisation on work tolerance and thermoregulation in trained tropical natives

This study aimed to investigate the effects of heat acclimatisation on thermoregulatory responses and work tolerance in trained individuals residing in the tropics. Eighteen male trained soldiers, who are native to a warm and humid climate, performed a total of four heat stress tests donning the Skeletal Battle Order (SBO, 20.5 kg) and Full Battle Order (FBO, 24.7 kg) before (PRE) and after (POST) a 10-day heat acclimatisation programme. The trials were conducted in an environmental chamber (dry bulb temperature: 32 °C, relative humidity: 70%, solar radiation: 400 W/m²). Excluding the data sets of which participants fully completed the heat stress tests (210 min) before and after heat acclimatisation, work tolerance was improved from 173±30 to 201±18 min (∼21%, p<0.05, n=9) following heat acclimatisation. Following heat acclimatisation, chest skin temperature during exercise was lowered in SBO (PRE=36.7±0.3 vs. POST=36.5±0.3 °C, p<0.01) and FBO (PRE=36.8±0.4 vs. POST=36.6±0.3 °C, p<0.01). Ratings of perceived exertion were decreased with SBO and FBO (PRE=11±2; POST=10±2; p<0.05) after heat acclimatisation. Heat acclimatisation had no effects on baseline body core temperature, heart rate and sweat rate across trials (p>0.05). A heat acclimatisation programme improves work tolerance with minimal effects on thermoregulation in trained tropical natives.     

An analytical model of temperature regulation in human head

An analytical model of human brain temperature regulation is proposed. The model describes the distribution of brain temperature as a function of internal and external parameters, such as temperature of the incoming arterial blood, blood flow, oxygen consumption rate, ambient temperature, and heat exchange with the environment. It is shown that substantial changes in human brain temperature can be accomplished only through changes in the temperature of the incoming arterial blood or substantial suppression of blood flow. Other parameters can lead only to temperature changes near the brain surface.     

舟山眼镜蛇对光暗周期加热光源反应所导致的体温变化

用 7条舟山眼镜蛇 (Najaatra)研究动物对光暗周期加热光源反应所导致的体温变化。设计两项实验 ,每项实验历时 9d。实验一光照期覆盖整个白天 ,实验二光照期覆盖整个晚上。眼镜蛇仅在加热光源开启期间进行体温调节 ,但光照期内任何阶段都未发现所有个体同时处于热活动状态。两项实验中热活动个体百分比的时间变化显著 ,实验二热活动个体百分比波动相对大于实验一。在加热光源开启期间 ,实验一热活动眼镜蛇的百分比总体上大于实验二。在两项实验中 ,热活动眼镜蛇体温的时间变化都不显著。实验一热活动眼镜蛇的体温高于实验二 ,而两项实验中不处于热活动状态的眼镜蛇的平均体温无显著差异。实验一热活动眼镜蛇 (31 1±0 8°C)选择的体温上限高于实验二眼镜蛇 (2 6 0± 0 9°C)。在两项实验的任何时间段内 ,眼镜蛇的体温都不低于环境温度。