Three-dimensional pore space quantification of apple tissue using X-ray computed microtomography

The microstructure and the connectivity of the pore space are important variables for better understanding of the complex gas transport phenomena that occur in plant tissues. In this study, we present an experimental procedure for image acquisition and image processing to quantitatively characterize in 3D the pore space of apple tissues (Malus domestica Borkh.) for two cultivars (Jonagold and Braeburn) taken from the fleshy part of the cortex using X-ray computer microtomography. Preliminary sensitivity analyses were performed to determine the effect of the resolution and the volume size (REV, representative elementary volume analysis) on the computed porosity of apple samples. For comparison among cultivars, geometrical properties such as porosity, specific surface area, number of disconnected pore volumes and their distribution parameters were extracted and analyzed in triplicate based on the 3D skeletonization of the pore space (medial axis analysis). The results showed that microtomography provides a resolution at the micrometer level to quantitatively analyze and characterize the 3D topology of the pore space in apple tissue. The computed porosity was confirmed to be highly dependent of the resolution used, and the minimum REV of the cortical flesh of apple fruit was estimated to be 1.3 mm³. Comparisons among the two cultivars using a resolution of 8.5 μm with a minimum REV cube showed that in spite of the complexity and variability of the pore space network observed in Jonagold and Braeburn apples, the extracted parameters from the medial axis were significantly different (P-value < 0.05). Medial axis parameters showed potential to differentiate the microstructure between the two evaluated apple cultivars.     

High-resolution X-ray microtomography for three-dimensional visualization of human stem cell muscle homing

In the perspective of clinical translation of stem cell research, it would be advantageous to develop new techniques to detect donor cells after transplantation to track their fate and thus better understand their role in regeneration of damaged and diseased tissues. In this study we use X-ray computed microtomography for three-dimensional visualization of stem cells that were labeled with magnetic nanoparticles and transplanted via intra-arterial infusion. We show that X-ray computed microtomography offers the possibility to detect with high definition and resolution human cells after transplantation, and opens new possibilities for both experimental stem cell research.     

内蒙古学者对蜘蛛的研究

蛛形纲蜘蛛目动物越来越受到当今动物学家的关注,从上世纪80年代开始,内蒙古学者对蜘蛛分类及区系方面进行了大量的研究,取得了不少成果,但是至今不曾有人对他们的研究成果进行过全面的总结。本文整理和分析了该地区学者30多年来发表的有关蜘蛛方面的40篇文献,介绍相关研究成果,试图给出一个全面的图景。     

Functional morphology of Tethya species (Porifera): 1. Quantitative 3D-analysis of Tethya wilhelma by synchrotron radiation based X-ray microtomography

Rhythmic body contraction is a phenomenon in the Porifera, which is only partly understood. As a foundation for the understanding of the functional morphology of the highly contractile Tethya wilhelma, we performed a qualitative and quantitative volumetric 3D-analysis of the morphology of a complete non-contracted specimen at resolutions of 5.2 and 6.9 μm, using synchrotron radiation based X-ray computed microtomography (SR-μCT). For the first time, we were able to visualize all three major body structures of a complete poriferan without dissection of the shock-frozen, fixed and contrasted specimen in a near-to-life confirmation: poriferan tissue, mineral skeleton and aquiferous system. Applying a ‘virtual cast’ technique allowed us to analyze the structural details of the complete canal structure. Our results imply an extensive re-circulation of water inside the poriferan due to well-developed by-pass-canals, connecting excurrent and incurrent system. Nevertheless, the oscule region is strictly separated from the incurrent system. Based on our data, we developed a hypothetical flow regime for T. wilhelma, which explains the necessity of by-pass canals to minimize pressure boosts in the canal system during contraction. Additionally, re-circulation optimizes nutrient uptake, within small-sized poriferans, like T. wilhelma. Quantitative analysis allowed us to measure volumes and surfaces, displaying remarkable organizational differences between choanosome and cortex, by means of distribution of morphological elements. The surface-to-volume ratio proved to be very high, underlining the importance of the poriferan pinacoderm. We support a pinacoderm-contraction hypothesis.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .Dedicated to Prof. Dr. Michele Sarà (Genova, Italy), in honour of his 80th birthday in 2006.     

Foraging advantages of mixed-species association between solitary and colonial orb-weaving spiders

This study investigated association between solitary orb-weaving spiders and a colonial orb-weaving spider, Metepeira incrassata (Araneae: Araneidae). Spiders were sampled along transects and an index of species association showed that two of the species were associated more frequently than expected based on a null hipothesis of random co-occurrence. The potential advantages of mixed-species association were investigated by comparing prey-capture success of one of these associates, Nephila clavipes (Araneae: Tetragnathidae), when it occurs alone, in single-species groups, and when associated with M. incrassata colonies. Field observations of prey-capture success by all three of these categories of N. clavipes revealed that individuals in M. incrassata colonies captured significantly more prey than solitaries or individuals in single-species groups. The increase in prey capture by N. clavipes in M. incrassata colonies may result from utilization of a foraging niche which intercepts a diffirent spectrum of prey than that available to single-species groups or solitaries. Related to this enhanced prey consumption is greater fecundity of spiders in association with M. incrassata as compared to solitaries or individuals in single-species groups.     

An X-ray microtomography-based method for detailed analysis of the three-dimensional morphology of fungal pellets

Filamentous fungi are widely used in the production of biotechnological compounds. Since their morphology is strongly linked to productivity, it is a key parameter in industrial biotechnology. However, identifying the morphological properties of filamentous fungi is challenging. Owing to a lack of appropriate methods, the detailed three-dimensional morphology of filamentous pellets remains unexplored. In the present study, we used state-of-the-art X-ray microtomography (µCT) to develop a new method for detailed characterization of fungal pellets. µCT measurements were performed using freeze-dried pellets obtained from submerged cultivations. Three-dimensional images were generated and analyzed to locate and quantify hyphal material, tips, and branches. As a result, morphological properties including hyphal length, tip number, branch number, hyphal growth unit, porosity, and hyphal average diameter were ascertained. To validate the potential of the new method, two fungal pellets were studied—one from Aspergillus niger and the other from Penicillium chrysogenum. We show here that µCT analysis is a promising tool to study the three-dimensional structure of pellet-forming filamentous microorganisms in utmost detail. The knowledge gained can be used to understand and thus optimize pellet structures by means of appropriate process or genetic control in biotechnological applications.     

Two-dimensional imaging detectors for structural biology with X-ray lasers

Our ability to harness the advances in microelectronics over the past decade(s) for X-ray detection has resulted in significant improvements in the state of the art. Biology with X-ray free-electron lasers present daunting detector challenges: all of the photons arrive at the same time, and individual high peak power pulses must be read out shot-by-shot. Direct X-ray detection in silicon pixel detectors—monolithic or hybrid—are the standard for XFELs today. For structural biology, improvements are needed for today''s 10–100 Hz XFELs, and further improvements are required for tomorrow''s 10+ kHz XFELs. This article will discuss detector challenges, why they arise and ways to overcome them, along with the current state of the art.     

The role of spider cocoons in controlling desiccation

Summary The abilities of the cocoons of the spiders Mecynogea lemniscata and Argiope aurantia to protect the enclosed egg and spiderling stages from desiccation were investigated in the laboratory under controlled humidities, and in the field under ambient conditions. For M. lemniscata, which has a relatively small clutch (8–30 eggs) and remains in the cocoon for approximately 9–10 months, removal of the cocoon had no effect on water loss from the egg stage, nor did it adversely affect hatching or molting success. Cocoon removal did, however, significantly affect water loss and, consequently, survival in the spiderling stage at all humidities in the laboratory and in the field. The importance of the cocoon for survival is probably related to the unusually long time M. lemniscata spiderlings spend in the cocoon overwintering. For A. aurantia, which has a substantially larger clutch size (300–1400 eggs) and remains in the cocoon for a shorter 6–7 months, cocoon removal had no effect on water loss, egg hatching success, molting success, nor spiderling survival. The lack of an effect suggests that other factors (e.g., relative humidity at the oviposition site, or a large clutch size) may be more important in controlling water loss for A. aurantia.     

Ecdysis period of Rhodnius prolixus head investigated using phase contrast synchrotron microtomography

Microtomography using synchrotron sources is a useful tool in biological imaging research since the phase coherence of synchrotron beams can be exploited to obtain images with high contrast resolution. This work is part of a series of works using phase contrast synchrotron microtomography in the study of Rhodnius prolixus head, the insect vector of Chagas’ disease, responsible for about 12,000 deaths per year. The control of insect vector is the most efficient method to prevent this disease and studies have shown that the use of triflumuron, a chitin synthesis inhibitor, disrupted chitin synthesis during larval development and it’s an alternative method against insect pests.The aim of this work was to investigate the biological effects of treatments with triflumuron in the ecdysis period (the moulting of the R. prolixus cuticle) using the new imaging beamline IMX at LNLS (Brazilian Synchrotron Light Laboratory). Nymphs of R. prolixus were taken from the Laboratory of Biochemistry and Physiology of Insects, Oswaldo Cruz Foundation, Brazil. Doses of 0.05 mg of triflumuron were applied directly to the abdomen on half of the insects immediately after feeding. The insects were sacrificed 25 days after feeding (intermoulting period) and fixed with glutaraldehyde.The results obtained using phase contrast synchrotron microtomography in R. prolixus showed amazing images of the effects of triflumuron on insects in the ecdysis period, and the formation of the new cuticle on those which were not treated with triflumuron. Both formation and malformation of this insect’s cuticle have never been seen before with this technique.     

The comparative study of the predatory behaviour of Myrmarachne, ant-like jumping spiders (Araneae: Salticidae)

The predatory behaviour of 31 species of Myrmarachne , ant-like salticids, was studied in the laboratory and the field. The ant-like morphology and locomotion of these spiders appears to function primarily in Batesian mimicry. No evidence was found of Myrmarachne feeding on ants. However, predatory sequences were found to differ considerably from those typical of salticids. Instead of stalking and leaping on prey, Myrmarachne lunged at prey from close range. Myrmarachne used its legs I to tap prey before lunging, another unusual behaviour for a salticid. Myrmarachne fed on a wide range of arthropod prey in nature and the laboratory, but appears to be especially efficient at catching moths. Also, Myrmarachne tends to open up, or enter into, other spiders' nests and eat other spiders' eggs. Myrmarachne males were less efficient than females, in laboratory tests, at catching various types of arthropod prey, but they appear to be as efficient as females at oophagy. Myrmarachne tend to use webs of other spiders as nest sites, but no evidence was found of Myrmarachne preying on spiders in webs. It appears that the unusual features of Myrmarachne's predatory and nesting behaviour are important in enabling these spiders to preserve their ant-like appearance.     

The diversity,recombination and horizontal transmission of Wolbachia in spiders in China

Wolbachia are maternally inherited endosymbiotic bacteria. These intracellular bacteria are common in arthropods and could manipulate host reproduction in diverse ways, such as feminization, parthenogenesis, male killing and cytoplasmic incompatibility. In spiders, infection by Wolbachia has been found in a total of 99 species belonging to 62 genera and 17 families. Furthermore, recent studies analyzed the phylogeny of Wolbachia in Hylyphantes graminicola, 2 cave spiders and Agelenopsis species using multilocus sequence typing (MLST) approach. However, the diversity of Wolbachia strains determined by MLST in spiders from China is still largely unknown.In this study, we collected 1153 spider individuals from Mangshan in China and screened for Wolbachia in 975 individuals representing 68 spider species belonging to 45 genera of 16 families. We analyzed the phylogenetic relationship between Wolbachia and their host spiders by MLST approach. We found novel infections of Wolbachia in 1 family, 9 genera and 20 species of spiders. We found 13 new Wolbachia strains and suggest that group A is more common than group B in Wolbachia that infect spiders. Our results revealed three recombination events of the concatenated multilocus sequences in Wolbachia that infect spiders. Furthermore, our results demonstrated the phylogenetic incongruence between Wolbachia and spiders, suggesting the horizontal transmission of Wolbachia in spiders.We suggest that recombination and horizontal transmission may play an important role in the diversity and evolution of Wolbachia in spiders.     

High quality 3D imaging of vertebrate microremains using X-ray synchrotron phase contrast microtomography

Vertebrate microremains, particularly teeth, represent a substantial part of known vertebrate biodiversity. Many groups, such as Mesozoic mammals, are known mostly through isolated teeth. Classical imaging techniques of such complex millimetric to inframillime-tric objects are most often limited by problems of manipulation, depth of focus or limited orientation. The methods generally used are stereomicroscopy (including in-focus z-series reconstruction) and Scanning Electron Microscopy (SEM), which provide good images. Nevertheless, both provide 2D static images or partial and directional 3D data, making complete observation difficult. Propagation phase contrast synchrotron X-ray microtomography is a powerful technique alleviating these limitations. Thanks to submicron resolution and to the edge detection effect, it rapidly provides 3D data from minute samples with levels of quality and detail unattainable using conventional microtomographs. Complex morphology of small specimens can be studied with unlimited orientation possibilities and, when coupled with 3D printing, it allows enlarged 3D reproductions of such small and fragile fossils.     

Failure Analysis of Batteries Using Synchrotron-based Hard X-ray Microtomography

Imaging morphological changes that occur during the lifetime of rechargeable batteries is necessary to understand how these devices fail. Since the advent of lithium-ion batteries, researchers have known that the lithium metal anode has the highest theoretical energy density of any anode material. However, rechargeable batteries containing a lithium metal anode are not widely used in consumer products because the growth of lithium dendrites from the anode upon charging of the battery causes premature cell failure by short circuit. Lithium dendrites can also form in commercial lithium-ion batteries with graphite anodes if they are improperly charged. We demonstrate that lithium dendrite growth can be studied using synchrotron-based hard X-ray microtomography. This non-destructive imaging technique allows researchers to study the growth of lithium dendrites, in addition to other morphological changes inside batteries, and subsequently develop methods to extend battery life.     

Does women's greater fear of snakes and spiders originate in infancy?

Previous studies with adult humans and nonhuman animals revealed more rapid fear learning for spiders and snakes than for mushrooms and flowers. The current experiments tested whether 11-month-olds show a similar effect in learning associative pairings between facial emotions and fear-relevant and fear-irrelevant stimuli. Consistent with the greater incidence of snake and spider phobias in women, results show that female but not male infants learn rapidly to associate negative facial emotions with fear-relevant stimuli. No difference was found between the sexes for fear-irrelevant stimuli. The results are discussed in relation to fear learning, phobias, and a specialized evolved fear mechanism in humans.     

The effect of food supply on the colonisation of barley by aerially dispersing spiders (Araneae)

A field experiment was carried out to determine whether different levels of food availability affected the retention rate of ballooning spiders landing in trays of seedling barley plants, half of which were infested with aphids from laboratory cultures. The trays were placed within bases in the field, then collected sequentially and spider numbers assessed in each tray. Deposition trays, containing trapping fluid only, were used to measure ballooning activity throughout the experimental period. The experiment was repeated four times. Overall, ballooning spiders were more likely to be retained in trays where aphid prey were present, with a total of 340 spiders found in the infested trays and 251 in the aphid-free trays, over the four experiments. Most of the spiders found were of the family Linyphiidae. In the second and fourth experiments the increased retention of spiders in the aphid-infested trays was statistically significant. Immature linyphiids alone also showed significantly higher retention in the infested trays in those two experiments and in the fourth experiment were largely responsible for the higher numbers found in the infested trays. There was also a statistically significant trend for a higher retention rate of female spiders, compared to males, in the barley trays than would have been expected from the ratios of females to males caught in the deposition trapping trays alone.     

Negative effects of ant foraging on spiders in Douglas-fir canopies

 Spiders and ants are potential competitors and mutual predators. Indirect evidence from previous research has suggested that ant foraging may significantly lower the abundance of arboreal spiders in young Douglas-fir plantations in western Oregon. This study tested the effect of foraging by ants, dominated by Camponotus spp., on spider assemblages in Douglas-fir canopies in a 5-month ant-exclusion experiment. The biomass of potential prey organisms on foliage, dominated by Psocoptera, increased significantly by 1.9- to 2.4-fold following ant exclusion. The removal of ants did not affect the abundance of flying arthropods in the vicinity of tree canopies as indicated by sticky trap catches. The abundance of hunting spiders, the majority being Salticidae, increased significantly by 1.5- to 1.8-fold in trees without ants in the late summer; neither the abundance of web-building spiders nor the average body size of hunting and web-building spiders were significantly affected by ant removal. Spider diversity and community structure did not differ significantly between control and ant-removal trees. The majority of prey captured by ants were Aphidoidea (48.1%) and Psocoptera (12.5%); spiders represented only 1.4% of the ants’ diet. About 40% of observed ants were tending Cinara spp. aphids. Our observations suggest that the lower abundance of hunting spiders in control canopies with ants may be due to interference competition with ants resulting from ant foraging and aphid-tending activities. Direct predation of spiders by ants appeared to be of minor importance in this study system. This study did not provide sufficient evidence for exploitative competition for prey between ants and spiders. Received: 21 February 1996 / Accepted: 14 August 1996