Reduced performance of prey targeting in pit vipers with contralaterally occluded infrared and visual senses

Both visual and infrared (IR) senses are utilized in prey targeting by pit vipers. Visual and IR inputs project to the contralateral optic tectum where they activate both multimodal and bimodal neurons. A series of ocular and pit organ occlusion experiments using the short-tailed pit viper (Gloydius brevicaudus) were conducted to investigate the role of visual and IR information during prey targeting. Compared with unoccluded controls, snakes with either both eyes or pit organs occluded performed more poorly in hunting prey although such subjects still captured prey on 75% of trials. Subjects with one eye and one pit occluded on the same side of the face performed as well as those with bilateral occlusion although these subjects showed a significant targeting angle bias toward the unoccluded side. Performance was significantly poorer when only a single eye or pit was available. Interestingly, when one eye and one pit organ were occluded on opposite sides of the face, performance was poorest, the snakes striking prey on no more than half the trials. These results indicate that, visual and infrared information are both effective in prey targeting in this species, although interference between the two modalities occurs if visual and IR information is restricted to opposite sides of the brain.     

Wide-band spectral tuning of heat receptors in the pit organ of the copperhead snake (Crotalinae)

Receptors located in the facial pit organ of certain species of snake signal the presence of prey. Infrared radiation is an effective stimulus suggesting that these receptors may be low-threshold temperature receptors. We recorded from the nerve innervating the pit organ of snakes belonging to the family of Crotalinae while stimulating the receptive area with well-defined optical stimuli. The objective was to determine the sensitivity of these receptors to a wide range (0.400-10.6 micro m) of optical stimuli to determine if a temperature-sensitive or photosensitive protein initiated signal transduction. We found that receptors in the pit organ exhibited a unique broad response to a wide range of electromagnetic radiation ranging from the near UV to the infrared. The spectral tuning of these receptors parallels closely the absorption spectra of water and oxyhemoglobin, the predominant chromophore in tissue. Our results support the hypothesis that these are receptors activated by minute temperature changes induced by direct absorption of optical radiation in the thin pit organ membrane.     

The Python pit organ: imaging and immunocytochemical analysis of an extremely sensitive natural infrared detector

The Python infrared-sensitive pit organ is a natural infrared imager that combines high sensitivity, ambient temperature function, microscopic dimensions, and self-repair. We are investigating the spectral sensitivity and signal transduction process in snake infrared-sensitive neurons, neither of which is understood. For example, it is unknown whether infrared receptor neurons function on a thermal or a photic mechanism. We imaged pit organs in living Python molurus and Python regius using infrared-sensitive digital video cameras. Pit organs were significantly more absorptive and/or emissive than surrounding tissues in both 3-5 microns and 8-12 microns wavelength ranges. Pit organs exhibited greater absorption/emissivity in the 8-12 microns range than in the 3-5 microns range. To directly test the relationship between photoreceptors and pit organ infrared-sensitive neurons, we performed immunocytochemistry using antisera directed against retinal photoreceptor opsins. Retinal photoreceptors were labeled with antisera specific for retinal opsins, but these antisera failed to label terminals of infrared-sensitive neurons in the pit organ. Infrared-receptive neurons were also distinguished from retinal photoreceptors on the basis of their calcium-binding protein content. These results indicate that the pit organ absorbs infrared radiation in two major atmospheric transmission windows, one of which (8-12 microns) matches emission of targeted prey, and that infrared receptors are biochemically distinct from retinal photoreceptors. These results also provide the first identification of prospective biochemical components of infrared signal transduction in pit organ receptor neurons.     

Behavioural examination of the infrared sensitivity of rattlesnakes (Crotalus atrox)

Pitvipers (Crotalinae) and boid snakes (Boidae) possess highly sensitive infrared (IR) receptors. The ability of these snakes to image IR radiation allows the assessment of the direction and distance of an IR source (such as warm-blooded prey) in the absence of visual cues. The aim of this study was to determine the behavioural threshold of snakes to an IR stimulus. A moving IR source of constant size and temperature was presented to rattlesnakes (Crotalus atrox) at various distances (10–160 cm) from their snout. The snakes’ responses were quantified by measuring distinct behavioural changes during stimulus presentation (head jerks, head fixed, freezing, rattling and tongue-flicking). The results revealed that C. atrox can detect an artificial IR stimulus resembling a mouse in temperature and size up to a distance of 100 cm, which corresponds to a radiation density of 3.35 × 10⁻³ mW/cm². These behavioural results reveal a 3.2 times higher sensitivity to IR radiation than earlier electrophysiological investigations.     

Investigation of the transduction mechanism of infrared detection in Melanophila acuminata: photo-thermal-mechanical hypothesis

Differential phase optical low coherence reflectometry (OLCR) was used to detect sub-wavelength displacements in the infrared-sensitive thoracic pit organ of Melanophila acuminata (Coleoptera: Buprestidae) upon absorption of infrared radiation at 3.39 microm. The displacement had more complex morphology but similar amplitude ( approximately 100 nm at 1 W cm(-2)) when compared to the displacement measured from the exocuticle in an alternate region on the beetle's body. In addition, a simplified finite difference model was developed to predict the temperature distribution and resultant thermal expansion in the pit organ tissue. The experimental and model results were interpreted to help clarify the mechanism by which the sensilla in the pit organ convert infrared radiation to neural signals. The results of this paper are discussed in relation to the photo-thermal-mechanical transduction hypothesis. This is the first experimental examination of the transduction mechanism in Melanophila acuminata.     

Eye and pit size are inversely correlated in crotalinae: Implications for selection pressure relaxation

Mate, prey, and predator recognition often depend on the integration of information from multiple sensory modalities including visual, auditory, and/or olfactory inputs. In Crotalinae, the eyes sense visible light while the pit organs detect infrared (IR) radiation. Previous studies indicate that there is significant overlap between the eye and pit sensory fields and that both senses are involved in recognition processes. This study investigated the relationships between eye and pit sizes in this taxonomic group as a function of phylogeny and habitat. In view of the fact that pit orientation depends largely on snout shape, pit vipers were grouped as follows: 1) arboreal, 2) terrestrial with rounded snout, and 3) terrestrial with pointed snout. The pit orientations and habitant patterns were fully independent of the Crotalinae phylogenetic tree. The phylogenetic generalized least squares model showed that both eye and pit areas were not of significantly phylogenetic relatedness, implying alternatively a strong effect of adaptation on eye and pit sizes. Negative correlations between relative eye and pit areas in terrestrial (both pointed and rounded snouts) and arboreal species were statistically significant. Our results suggest that the eyes and pits function in a complementary fashion such that selection for IR‐perception relaxes selection pressures on the visual system and selection for visual discrimination relaxes selection pressures acting on the IR‐system. J. Morphol. 277:107–117, 2016. © 2015 Wiley Periodicals, Inc.     

Modelling a historic oil-tank fire allows an estimation of the sensitivity of the infrared receptors in pyrophilous Melanophila beetles

Pyrophilous jewel beetles of the genus Melanophila approach forest fires and there is considerable evidence that these beetles can detect fires from great distances of more than 60 km. Because Melanophila beetles are equipped with infrared receptors and are also attracted by hot surfaces it can be concluded that these infrared receptors are used for fire detection.The sensitivity of the IR receptors is still unknown. The lowest threshold published so far is 0.6 W/m(2) which, however, cannot explain the detection of forest fires by IR radiation from distances larger than approximately 10 km. To investigate the possible sensitivity of the IR receptors we assumed that beetles use IR radiation for remote fire detection and we made use of a historic report about a big oil-tank fire in Coalinga, California, in 1924. IR emission of an oil-tank fire can be calculated by "pool fire" simulations which now are used for fire safety and risk analysis. Assuming that beetles were lured to the fire from the nearest forests 25 and 130 km away, our results show that detection from a distance of 25 km requires a threshold of the IR receptors of at least 3×10(-2) W/m(2). According to our investigations most beetles became aware of the fire from a distance of 130 km. In this case the threshold has to be 1.3×10(-4) W/m(2). Because such low IR intensities are buried in thermal noise we suggest that the infrared sensory system of Melanophila beetles utilizes stochastic resonance for the detection of weak IR radiation. Our simulations also suggest that the biological IR receptors might be even more sensitive than uncooled technical IR sensors. Thus a closer look into the mode of operation of the Melanophila IR receptors seems promising for the development of novel IR sensors.     

Cleaning Up After a Meal: The Consequences of Prey Disposal for Pit‐Building Antlion Larvae

Predators use a variety of strategies for capturing prey. Trap‐building predators can save on searching and encountering costs by investing in the construction and maintenance of traps such as webs and pits. However, what to do with partially consumed prey poses a potential problem. Antlion larvae (Myrmeleon acer) catch ants in conical pits, and dispose of partially consumed carcasses by flicking them a short distance away. We tested whether this prey‐disposal behaviour affects the effectiveness of antlion pits. We observed ant behaviour around artificially constructed pits and compared falls into pits with clean margins to those with conspecific ant carcasses or control objects around the pit edge. The presence of objects near pits affected the behaviour of live ants, and reduced the effectiveness of pits. Live ants spent the most time examining fresh ant carcasses, but the presence of any object near pits deterred pitfalls. Ants fell into pits significantly more often when pit edges were clean, suggesting that antlions could incur a prey capture cost from flicking carcasses from pits as well as from the accumulation of other debris around pit margins.     

Surface ultrastructure of pit organ, spectacle, and non pit organ epidermis of infrared imaging boid snakes: A scanning probe and scanning electron microscopy study.

Boid snakes possess unique infrared imaging pit organs. The ultrastructure of the surfaces of these organs scatter or reflect electromagnetic radiation of specific wavelengths. Pit organ epidermal surfaces of boid snakes are covered with arrays of pore-like structures called micropits. In order to determine the dimensions of this complicated surface structure, we have performed the first ultrastructural analysis on snake epidermis by high-resolution microscopy techniques. Using scanning probe microscopy and scanning electron microscopy, we found that the epidermis of pit organ, maxillary non pit organ, spectacle, and ventral scales contain arrays of micropits. These scale surfaces also contain major surface features of overlapping plate-like structures. Pit organ micropits averaged 319 nm in diameter and 46 nm in depth and were spaced an average of 808 nm from each other. These micropits were significantly deeper, of greater diameter, and spaced at greater distances apart than those of the other scales. Plate structures of the pit organs had a mean distance between plates of 3.5 microm and a mean plate step height of 151 nm. These differences serve to strengthen the argument that arrays of micropit and plate surface structures function as spectral filters or anti-reflective coatings with respect to incident electromagnetic radiation.     

Infrared-assisted tryptic proteolysis for peptide mapping

In this report, infrared (IR) radiation was employed to enhance the efficiency of tryptic proteolysis for peptide mapping. Protein solutions containing trypsin in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of BSA and myoglobin (MYO) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF MS with the sequence coverages of 69% (BSA) and 90% (MYO) that were much better than those obtained by conventional in-solution tryptic digestion. The present IR-assisted proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.     

Sensitivity threshold and response characteristics of infrared detection in the beetle Melanophila acuminata (Coleoptera: Buprestidae)

The minimum detection threshold of the infrared sensitive beetle, Melanophila acuminata, was measured with a helium-neon laser that emitted light at a wavelength of 3.39 microm. Extracellular recordings were taken both at the pit organ responsible for detection and at the interganglionic connectives in the thorax of the beetle. At the pit organ, generator and action potentials from single neurons were measured with a sharpened tungsten electrode. At the connectives that linked the fused second meso-/metathoracic and prothoracic ganglia, compound action potentials were measured with a tungsten hook electrode that encircled the connective. The latter recordings confirmed conveyance of infrared information through specific pathways to rostrally-situated sites in the nervous system of the beetle. The 50% probability irradiance threshold at which action potentials were elicited from the receptor and connectives occurred at 17.3 and 14.6 mW/cm(2), respectively. In addition to sensitivity threshold, several other characteristics of the response were quantified including dependence of generator potential latency, generator potential duration, spike frequency, and spike latency on irradiance, dependence of response strength (spike count) on exposure time, and flicker fusion frequency. The ability to detect infrared radiation is rare in nature, and these results provide valuable information necessary to understand this unique sensitivity.     

Evidence for localized cell heating induced by infrared optical tweezers.

The confinement of liposomes and Chinese hamster ovary (CHO) cells by infrared (IR) optical tweezers is shown to result in sample heating and temperature increases by several degrees centigrade, as measured by a noninvasive, spatially resolved fluorescence detection technique. For micron-sized spherical liposome vesicles having bilayer membranes composed of the phospholipid 1,2-diacyl-pentadecanoyl-glycero-phosphocholine (15-OPC), a temperature rise of approximately 1.45 +/- 0.15 degrees C/100 mW is observed when the vesicles are held stationary with a 1.064 microns optical tweezers having a power density of approximately 10(7) W/cm2 and a focused spot size of approximately 0.8 micron. The increase in sample temperature is found to scale linearly with applied optical power in the 40 to 250 mW range. Under the same trapping conditions, CHO cells exhibit an average temperature rise of nearly 1.15 +/- 0.25 degrees C/100 mW. The extent of cell heating induced by infrared tweezers confinement can be described by a heat conduction model that accounts for the absorption of infrared (IR) laser radiation in the aqueous cell core and membrane regions, respectively. The observed results are relevant to the assessment of the noninvasive nature of infrared trapping beams in micromanipulation applications and cell physiological studies.     

Infrared spectral sensitivity of Melanophila acuminata

The spectral sensitivity of the pit organ of the beetle Melanophila acuminata (Coleoptera:Buprestidae) was measured using an ultrafast tunable infrared laser source and standard electrophysiological techniques. The pit organ may be classified as a broadband detector as the beetles responded to all infrared excitation wavelengths from 2 to 6&mgr;m. There was a decrease in response threshold and latency and an increase in the magnitude of the response in the region from 2.8 to 3.5&mgr;m, which corresponded to a region of decreased transmittance (increased absorbance) as measured by Fourier transform infrared spectroscopy. The implications of the correlation between spectral response and optical properties are discussed.     

The effect of temperature on the functional response of Phytoseiulus persimilis (Acari: Phytoseiidae)

Environmental variables, such as temperature, are important in determining the efficiency of biological control in ornamental crops. This paper examines the effect of temperature on the functional response of adult female Phytoseiulus persimilis to eggs of the spider mite, Tetranychus urticae. The functional response was determined using a new functional response assay technique with plant stems as an arena, rather than leaf discs. The use of plant stems allows the influence that plant structure has on predation to be incorporated into the assay. Control assays were also used (without predators) to estimate natural losses of prey. The data were analysed using a binomial model, with the use of Abbot's formula to correct for the losses in the controls. A combined equation to describe the effect of temperature and prey density on the predation rate of Phytoseiulus persimilis was derived. The results showed that more prey are eaten as the temperature increases from 15 degrees C to 25 degrees C, but the number of prey eaten then declines at 30 degrees C, although not to the levels seen at 20 degrees C. The implication of these results for biological control in ornamental crops, where the temperature can often exceed 30 degrees C, is discussed.     

Biological applications of synchrotron radiation infrared spectromicroscopy

Extremely brilliant infrared (IR) beams provided by synchrotron radiation sources are now routinely used in many facilities with available commercial spectrometers coupled to IR microscopes. Using these intense non-thermal sources, a brilliance two or three order of magnitude higher than a conventional source is achievable through small pinholes (< 10 μm) with a high signal to-noise ratio. IR spectroscopy is a powerful technique to investigate biological systems and offers many new imaging opportunities. The field of infrared biological imaging covers a wide range of fundamental issues and applied researches such as cell imaging or tissue imaging. Molecular maps with a spatial resolution down to the diffraction limit may be now obtained with a synchrotron radiation IR source also on thick samples. Moreover, changes of the protein structure are detectable in an IR spectrum and cellular molecular markers can be identified and used to recognize a pathological status of a tissue. Molecular structure and functions are strongly correlated and this aspect is particularly relevant for imaging. We will show that the brilliance of synchrotron radiation IR sources may enhance the sensitivity of a molecular signal obtained from small biosamples, e.g., a single cell, containing extremely small amounts of organic matter. We will also show that SR IR sources allow to study chemical composition and to identify the distribution of organic molecules in cells at submicron resolution is possible with a high signal-to-noise ratio. Moreover, the recent availability of two-dimensional IR detectors promises to push forward imaging capabilities in the time domain. Indeed, with a high current synchrotron radiation facility and a Focal Plane Array the chemical imaging of individual cells can be obtained in a few minutes. Within this framework important results are expected in the next years using synchrotron radiation and Free Electron Laser (FEL) sources for spectro-microscopy and spectral-imaging, alone or in combination with Scanning Near-field Optical Microscopy methods to study the molecular composition and dynamic changes in samples of biomedical interest at micrometric and submicrometric scales, respectively.     

The challenge of predicting temperature effects on short-term predator–prey dynamics

Understanding how interacting species respond to changing temperatures is complicated because both inter- and intraspecific processes can be temperature-dependent, and because the dynamics of interest often occur over short time scales. While efforts to document and integrate inter- and intraspecific effects are ongoing, few broad conclusions have emerged. Here, we use a simple structured predator–prey model to explore whether some such conclusions can be reached with incomplete knowledge about initial population sizes or about how temperature alters predation rates. We find that, despite strong sensitivity of quantitative predictions to the model’s details, qualitative predictions for how predator–prey communities respond to temperature may still be possible. As a case study, we ask whether biological control of the herbivorous insect pea aphid will be strengthened or weakened by increasing temperatures. Our empirically derived parameter estimates suggest biological control will be strengthened, but we caution that this result is not inevitable given what we currently know about the effects of temperature in this system. Our work highlights the complex interaction between inter- and intraspecific effects of temperature in ecological communities.     

Temperature influences on functional response of Coenosia attenuata (Diptera: Muscidae) individuals

The functional response of single Coenosia attenuta Stein (Diptera: Muscidae) adult females to Drosophila melanogaster (Meigen) adults has been investigated at different but constant temperatures and prey densities. The experiments were conducted in cages over a wide range of temperatures between 12 and 42 degrees C. At each experimental temperature, the number of attacks increases with increasing prey density up to a maximum attack rate referred to as demand rate. A ratio-dependent and demand-driven functional response model was appropriate to describe the attack rate at different temperatures. Both the demand and the search rate linearly increased with increasing temperatures throughout the range. Based on these results, a temperature- and prey density-dependent bidimensional attack rate model was developed and parameterized. The resulting response surface shows that C. attenuata is active over a wide range of temperatures (from approximately 12 to 36 degrees C) and attacks occur up to 42 degrees C. Thus, C. attenuata may be a promising biological control agent in Mediterranean greenhouses. To support this conclusion, further studies including the evaluation of spatial scale effects on the search rate and the consideration of the economically relevant whitefly prey are recommended.     

Sand as a medium for transmission of vibratory signals of prey in antlions Euroleon nostras (Neuroptera: Myrmeleontidae)

Abstract.  European pit-building antlions ( Euroleon nostras / Geoffroy in Fourcroy/) detect their prey by sensing the vibrations that prey generate during locomotory activity. The behavioural reactions and some of the physical properties of substrate vibrations in sand are measured to observe signal transmission through the substrate. The frequency range of the signals of four arthropod species ( Tenebrio molitor , Pyrrhocoris apterus , Formica sp. and Trachelipus rathkei ) is 0.1–4.5 kHz and acceleration values are in the range 400 μm s⁻² to 1.5 mm s⁻². Substrate particle size and the frequency of prey signals both influence the propagation properties of vibratory signals. The damping coefficient at a frequency 300 Hz varies from 0.26 to 2.61 dB cm⁻¹ and is inversely proportional to the size of the sand particle. The damping coefficient is positively correlated with the frequency of the pulses. Vibrations in finer sand are attenuated more strongly than in coarser sand and, consequently, an antlion detects its prey only at a short distance. The reaction distance is defined as the distance of the prey from the centre of the pit when the antlion begins tossing sand as a reaction to the presence of prey. The mean reaction distance is 3.3 cm in the finest sand (particle size ≤ 0.23 mm) and 12.3 cm in coarser sand (particle size 1–1.54 mm). The most convenient sands for prey detection are considered to be medium particle-sized sands.     

Efficient chymotryptic proteolysis enhanced by infrared radiation for peptide mapping

Infrared (IR) radiation was employed to enhance the efficiency of chymotryptic proteolysis for peptide mapping in this work. Protein solutions containing chymotrypsin in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. BSA and cytochrome c (Cyt- c) were digested by IR-assisted chymotryptic proteolysis to demonstrate the feasibility and performance of the novel digestion approach and the digestion time was significantly reduced to 5 min. The obtained digests were further identified by MALDI-TOF MS with the sequence coverages that were comparable to those obtained by using conventional in-solution digestion. The suitability of IR-assisted chymotryptic proteolysis to complex proteins was demonstrated by digesting human serum. The present proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.     

Analysis of coated pit recycling on human fibroblasts

The recycling to the cell surface of previously internalized coated pits has been proposed as a likely mechanism for the rapid regeneration of coated pits on human fibroblast surfaces at 37 degrees C (1). We present a general mathematical model of coated pit recycling for the case when the coat cycles as a single unit, and use it to analyze certain time and temperature dependent data obtained by Anderson et al. (1) and Vermeer et al. (2). We show how recycling can account for these data and how this type of data can be used to distinguish between different possible recycling mechanisms. We show that these data are inconsistent with a two compartment model where coat material simply shuttles back and forth between coated pits and short-lived coated vesicles. From these data we estimate for human fibroblasts at 37 degrees C: that the time for a coated pit to be replenished through recycling after it is lost through internalization is greater than 3.5 min; and that at any moment 53% or less of the cell's clathrin that is involved in coated pit recycling is on the cell surface.