Gerald Kastberger

Infrared imaging technology and biological applications

Temperature is the most frequently measured physical quantity, second only to time. Infrared (IR) technology has been utilized successfully in astronomy (for a summary, see Hermans-Killam, 2002b) and in industrial and research settings (Gruner, 2002; Madding, 1982, 1989; Wolfe & Zissis, 1993) for decades. However, fairly recent innovations have reduced costs, increased reliability, and resulted in noncontact IR sensors offering mobile, smaller units of measurement (EOI, 2002; Flir, 2000, 2001, 2002). The advantages of using IR imaging are (1) rapidity in the millisecond range, facilitating measurement of moving targets, (2) noncontact procedures, allowing measurements of hazardous or physically inaccessible objects, (3) no interference and no energy lost from the target, (4) no risk of contamination, and (5) no mechanical effect on the surface of the object. All these factors have led to IR technology’s becoming an area of interest for new kinds of applications and users. In both manufacturing and quality control, temperature plays an important role as an indicator of the condition of a product or a piece of machinery (EOI, 2002; Flir, 2000, 2001, 2002; Raytek, 2002). In medical and veterinary applications, IR thermometry is increasingly used in organ diagnostics, in the evaluation of sports injuries and the progression of therapy, in disease evaluation (e.g., breast cancer, arthritis, and SARS; Flir, 2003), and in injury and inflammation examinations in horses, livestock (Tivey & Banhazi, 2002), and zoo animals (Hermans-Killam, 2002a; Thiesbrummel, 2002). Lastly, physiological expressions of life processes in animals (Kastberger, Winder, & Steindl, 2001; Stabentheiner, Kovac, & Hagmüller, 1995; Stabentheiner, Kovac, & Schmaranzer, 2002; Stabentheiner & Schmaranzer, 1987) and plants (Bermadinger-Stabentheiner & Stabentheiner, 1995) can be monitored. The most recent field in which IR technology has been applied is animal behavior. This article focuses on the practical options for noncontact IR thermometry— in particular, in biological applications.

Noninvasive diagnosis of seed viability using infrared thermography

Recent advances in the noninvasive analyses of plant metabolism include stress imaging techniques, mainly developed for vegetative tissues. We explored if infrared thermography can be used to predict whether a quiescent seed will germinate or die upon water uptake. Thermal profiles of viable, aged, and dead Pisum sativum seeds were recorded, and image analysis of 22,000 images per individual seed showed that infrared thermography can detect imbibition- and germination-associated biophysical and biochemical changes. These “thermal fingerprints” vary with viability in this species and in Triticum aestivum and Brassica napus seeds. Thermogenesis of the small individual B. napus seeds was at the limit of the technology. We developed a computer model of “virtual pea seeds,” that uses Monte Carlo simulation, based on the heat production of major seed storage compounds to unravel physico-chemical processes of thermogenesis. The simulation suggests that the cooling that dominates the early thermal profiles results from the dissolution of low molecular-weight carbohydrates. Moreover, the kinetics of the production of such “cooling” compounds over the following 100 h is dependent on seed viability. We also developed a deterministic tool that predicts in the first 3 hours of water uptake, when seeds can be redried and stored again, whether or not a pea seed will germinate. We believe that the early separation of individual, ungerminated seeds (live, aged, or dead) before destructive germination assessment creates unique opportunities for integrative studies on cell death, differentiation, and development.

Giant honeybees return to their nest sites.

The Asian giant honeybee Apis dorsata forms massive single-comb colonies which usually hang from a tree branch or the eaves of buildings. Although colonies regularly migrate over many kilometres, we find that they often return to their original nest site — even after an absence of up to two years. How the bees do this is unknown, as workers live for only a few weeks.

Small hive beetles survive in honeybee prisons by behavioural mimicry

Abstract. We report the results of a simple experiment to determine whether honeybees feed their small hive beetle nest parasites. Honeybees incarcerate the beetles in cells constructed of plant resins and continually guard them. The longevity of incarcerated beetles greatly exceeds their metabolic reserves. We show that survival of small hive beetles derives from behavioural mimicry by which the beetles induce the bees to feed them trophallactically. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at htpp://dx.doi.org/10.1007/s00114-002-0326-y.

Spotted hyenas (Crocuta crocuta) follow migratory prey. Seasonal expansion of a clan territory in Etosha, Namibia

The spatial organization of one clan of spotted hyenas Crocuta crocuta in the centre of the Etosha National Park, Namibia, is described during the dry and the wet seasons. The clan comprised 11 adults and sub-adults and occupied a territory of 160 km2 in the dry season and 320 km2 in the wet season. The dry season territory contained a low density (one animal/km2) of resident herbivores, such as gemsbok Oryx gazella, kudu Tragelaphus strepsiceros, giraffe Giraffa camelopardis, steenbok Raphicerus campestris and ostrich Struthio camelus, and a higher density of migratory species (12 animals/km2), principally springbok Antidorcas marsupialis, zebra Equus burchelli and wildebeest Connochaetes taurinus. These migratory species were the main prey of clan members. At the start of the wet season, the migratory herbivores migrated to the north-west, resulting in a considerable decline in the density of prey in the area used by clan members during the dry season. In response to this decline in prey, clan members followed the migratory herds and shifted the focus of their activities to an area grazed by migratory herbivores during the wet season. There existed a strong spatial relationship between the hyena density and the migratory prey density in both the dry and wet season. We postulate that the considerable enlargement of the clan territory in the wet season is a response to both the migratory movements of prey and an increase in the dispersion of prey during the wet season.

Aggressive and Docile Colony Defence Patterns in Apis mellifera. A Retreater-Releaser Concept

Colony defence in Apis mellifera involves a variety of traits ranging from ‘aggressive’ (e.g. entrance guarding, recruitment of flying guards) to ‘docile’ (e.g. retreating into the nest) expression. We tested 11 colonies of three subspecies (capensis, scutellata, carnica) regarding their defensiveness. Each colony was selected as reportedly ‘aggressive’, ‘intermediate’ or ‘docile’ and consisted of about 10,000 bees. We applied three stimulation regimes (mechanical disturbance, exposure to alarm pheromones, and the combination of both) and measured their behaviours by tracking the rates of outflying bees at the entrance sites of the test hives. We provided evidence that for mechanical disturbances the test colonies resolved into two response types, if the ‘immediate’ defence response, assessed in the first minute of stimulation, was taken as a function of foraging: ‘releaser’ colonies allocated flying guards, ‘retreater’ colonies reduced the outside-hive activities. This division was observed irrespective of the subspecies membership and maintained in even roughly changing environmental conditions. However, if pheromone and mechanical stimulation were combined, the variety of colony defensiveness restricted to two further types irrespective of the subspecies membership: six of nine colonies degraded their rate of flying defenders with increasing foraging level, three of the colonies extended their ‘aggressiveness’ by increasing the defender rate with the foraging level. Such ‘super-aggressive’ colonies obviously are able to allocate two separate recruitment pools for foragers and flying defenders.

‘Special agents’ trigger social waves in giant honeybees (Apis dorsata)

Giant honeybees (Apis dorsata) nest in the open and have therefore evolved a variety of defence strategies. Against predatory wasps, they produce highly coordinated Mexican wavelike cascades termed ‘shimmering’, whereby hundreds of bees flip their abdomens upwards. Although it is well known that shimmering commences at distinct spots on the nest surface, it is still unclear how shimmering is generated. In this study, colonies were exposed to living tethered wasps that were moved in front of the experimental nest. Temporal and spatial patterns of shimmering were investigated in and after the presence of the wasp. The numbers and locations of bees that participated in the shimmering were assessed, and those bees that triggered the waves were identified. The findings reveal that the position of identified trigger cohorts did not reflect the experimental path of the tethered wasp. Instead, the trigger centres were primarily arranged in the close periphery of the mouth zone of the nest, around those parts where the main locomotory activity occurs. This favours the ‘special-agents’ hypothesis that suggest that groups of specialized bees initiate the shimmering.

Self-Assembly Processes in Honeybees: The Phenomenon of Shimmering

Giant honeybees, Apis dorsata, have evolved a plethora of defence behaviours that enable this lifestyle. Against predatory wasps, they display Mexican wave-like cascades of “shimmering” whereby hundreds of bees flip their abdomens upwards in a split second. Shimmering is a type of social defence that also has relevance for task partitioning, collective decision making and social communication. Shimmering also relies on unexplored principles of information transfer and is a compelling example of self-organisation. It addresses “proximate” questions (how does shimmering work?) and ultimate questions (why has shimmering evolved?) and, therefore, gives rise to the discovery of interdependencies between giant honeybees as prey and its predators as part of a “co-evolutionary arms race”. Here, we review the underlying mechanisms of the phenomenon of shimmering and prove some of its anti-predatory goals.

Isoelectric focusing as a tool to evaluate carabid beetles as predatory agents of the pest slug Arion lusitanicus

Isoelectric focusing was investigated to detect esterases of the pest slug Arion lusitanicus in the crop contents of predatory carabid beetles. The method is exceptionally well suited for field studies, as it is fast, cheap, easy to apply, and almost species-specific. The identification of A. lusitanicus was enabled by four characteristic, stable esterase electromorphs. The profile clearly differed from the band patterns of the digestive enzymes of the carabid beetles, from esterases of other potential prey species and even from the enzymes of early developmental stages of the slug. To test the heaviest stained isoenzyme for its decay under controlled conditions, three carabid species were fed on a fixed amount of slug hepatopancreas. In Carabus cancellatus and Pterostichus melanarius the enzyme lost 35% of its activity after 16 h, fitting a logarithmic curve, whereas in Carabus granulatus the same decay status was already reached after 8 h following a linear regression. Both, the similarities and differences between beetle species might be due to their mode of extra- versus intraintestinal digestion and the amount of hepatopancreas fed. The temperature during digestion was also influential, as the regression slopes of esterase activity between the two temperature regimes tested in C. granulatus were significantly different. The volume of crop contents decreased during digestion in both Carabus species, corresponding to the decline in esterase. In contrast, the crop volume of P. melanarius reached its maximum almost 4 h after ingestion. This was interpreted as an effect of its mode of ingestion and time-elapsed enzyme production.

How to Join a Wave: Decision-Making Processes in Shimmering Behavior of Giant Honeybees (Apis dorsata)

Shimmering is a collective defence behaviour in Giant honeybees (Apis dorsata) whereby individual bees flip their abdomen upwards, producing Mexican wave-like patterns on the nest surface. Bucket bridging has been used to explain the spread of information in a chain of members including three testable concepts: first, linearity assumes that individual “agent bees” that participate in the wave will be affected preferentially from the side of wave origin. The directed-trigger hypothesis addresses the coincidence of the individual property of trigger direction with the collective property of wave direction. Second, continuity describes the transfer of information without being stopped, delayed or re-routed. The active-neighbours hypothesis assumes coincidence between the direction of the majority of shimmering-active neighbours and the trigger direction of the agents. Third, the graduality hypothesis refers to the interaction between an agent and her active neighbours, assuming a proportional relationship in the strength of abdomen flipping of the agent and her previously active neighbours. Shimmering waves provoked by dummy wasps were recorded with high-resolution video cameras. Individual bees were identified by 3D-image analysis, and their strength of abdominal flipping was assessed by pixel-based luminance changes in sequential frames. For each agent, the directedness of wave propagation was based on wave direction, trigger direction, and the direction of the majority of shimmering-active neighbours. The data supported the bucket bridging hypothesis, but only for a small proportion of agents: linearity was confirmed for 2.5%, continuity for 11.3% and graduality for 0.4% of surface bees (but in 2.6% of those agents with high wave-strength levels). The complimentary part of 90% of surface bees did not conform to bucket bridging. This fuzziness is discussed in terms of self-organisation and evolutionary adaptedness in Giant honeybee colonies to respond to rapidly changing threats such as predatory wasps scanning in front of the nest.