Avian colour vision and avian video playback experiments

Video playback potentially allows the presentation, manipulation, and replication of realistic moving visual stimuli, in a way that is impossible with real animals or static dummies, and difficult even with mechanical models. However, there are special problems attached to the use of this technology; this article concentrates on the problem of accurate colour rendition. Video and television simulate the colour of objects rather than reproduce the spectrum of light that they naturally emit, transmit, or reflect. This simulation is achieved by using relatively narrow waveband light to stimulate the cone cells in the retina in a similar pattern to that produced by the natural object. However, species differ in the spectral tuning of their photoreceptors, so a faithful colour rendition for a human is unlikely to be achieved for another species. This problem is discussed with special reference to birds, a taxon renown for its colourfulness and frequent use in behavioural experiments but which has a very different colour vision from that of humans. We stress that the major pitfalls that can arise when using video playback with avian subjects can also occur in ’normal’ behavioural experiments. However, the problems of faithful colour rendition are particularly severe with video, and the major benefits that the technology brings will only be realised under a limited range of circumstances, with careful validation experiments. Received: 24 January 2000 / Received in revised form: 28 March 2000 / Accepted: 1 April 2000     

Response of fiddler crabs (Uca tangeri) to video playback in the field

The fiddler crab Uca tangeri communicates using a visual waving display and a vibratory drumming signal, both thought to function in mate attraction. Using video playback techniques, images of an empty mudflat, a waving male, a threatening male, and a wandering female were presented to male subjects. All stimuli elicited similar levels of low-intensity waving, but significantly more high-intensity waves were elicited by the female and threatening male stimuli than the mudflat stimulus or the waving male stimulus. This concurs with other research that the waving display is used at a higher intensity to attract females. The threatening male stimulus failed to elicit the same response as an actual threatening male and we discuss the likely reasons for this. The subjects also spent a significantly greater proportion of time drumming during the male waving stimulus than during the female stimulus, suggesting that drumming functions in male-male competition as well as female attraction. Received: 18 October 1999 / Received in revised form: 25 November 1999 / Accepted: 10 December 1999     

Design considerations and techniques for constructing video stimuli

Techniques for constructing video playback stimuli fall into five categories. The first three involve manipulating video sequences: (1) edited video is a temporal rearrangement of raw footage, (2) processed video applies global filtering algorithms to edited video, and (3) frame-manipulated video involves manually altering individual frames. The last two, (4) exemplar-based animation and (5) parameter-based animation, are synthetic models derived from visual parameters based on a single exemplar and sample data, respectively. Image-based approaches are straightforward to apply and preserve fine spatiotemporal detail. Synthetic stimuli are desirable when a large number of manipulations are called for and to ensure individual stimuli reflect population characteristics. Received: 13 December 1999 / Received in revised form: 25 February 2000 / Accepted: 1 March 2000     

Considerations on the use of video playbacks as visual stimuli: the Lisbon workshop consensus

This paper is the consensus of a workshop that critically evaluated the utility and problems of video playbacks as stimuli in studies of visual behavior. We suggest that video playback is probably suitable for studying motion, shape, texture, size, and brightness. Studying color is problematic because video systems are specifically designed for humans. Any difference in color perception must lead to a different color sensation in most animals. Another potentially problematic limitation of video images is that they lack depth cues derived from stereopsis, accommodation, and motion parallax. Nonetheless, when used appropriately, video playback allows an unprecedented range of questions in visual communication to be addressed. It is important to note that most of the potential limitations of video playback are not unique to this technique but are relevant to all studies of visual signaling in animals. Received: 20 October 1999 / Accepted: 5 November 1999     

Considerations in video playback design: using optic flow analysis to examine motion characteristics of live and computer-generated animation sequences

The increasing use of the video playback technique in behavioural ecology reveals a growing need to ensure better control of the visual stimuli that focal animals experience. Technological advances now allow researchers to develop computer-generated animations instead of using video sequences of live-acting demonstrators. However, care must be taken to match the motion characteristics (speed and velocity) of the animation to the original video source. Here, we presented a tool based on the use of an optic flow analysis program to measure the resemblance of motion characteristics of computer-generated animations compared to videos of live-acting animals. We examined three distinct displays (tail-flick (TF), push-up body rock (PUBR), and slow arm wave (SAW)) exhibited by animations of Jacky dragons (Amphibolurus muricatus) that were compared to the original video sequences of live lizards. We found no significant differences between the motion characteristics of videos and animations across all three displays. Our results showed that our animations are similar the speed and velocity features of each display. Researchers need to ensure that similar motion characteristics in animation and video stimuli are represented, and this feature is a critical component in the future success of the video playback technique.     

Flow cytometric analysis of the chromosomes and stability of a wheat cell-culture line

A rapidly growing, long-term suspension culture derived from Triticum aestivum L. (wheat) was synchronized using hydroxyurea and colchicine, and a chromosome suspension with chromosomes was made. After staining with the DNA-specific fluorochromes Hoechst 33258 and Chromomycin univariate and bivariate flow-cytometry histograms showed 15 clearly resolved peaks corresponding to individual chromosome types or groups of chromosomes with similar DNA contents. The flow karyotype was closely similar to a histogram of DNA content measurements of Feulgen-stained chromosomes made by microdensitometry. We were able to show the stability of the flow karyotype of the cell line over a year, while a parallel subculture had a slightly different, stable, karyotype following different growth conditions. The data indicate that flow cytometric analysis of plant karyotypes enables accurate, statistically precise chromosome classification and karyotyping of cereals. There was little overlap between individual flow-histogram peaks, so the method is useful for flow sorting and the construction of chromosome specific-recombinant DNA libraries. Using bivariate analysis, the AT:GC ratio of all the chromosomes was remarkably similar, in striking contrast to mammalian flow karyotypes. We speculate about a fundamental difference in organization and homogenization of DNA sequences between chromosomes within mammalian and plant genomes. Received: 24 April 1996 / Accepted: 24 May 1996     

Microsatellites in Zea - variability,patterns of mutations,and use for evolutionary studies

To evaluate the performance of microsatellites or simple sequence repeats (SSRs) for evolutionary studies in Zea, 46 microsatellite loci originally derived from maize were applied to diverse arrays of populations that represent all the diploid species of Zea and 101 maize inbreds. Although null phenotypes and amplification of more than two alleles per plant were observed at modest rates, no practical obstacle was encountered for applying maize microsatellites to other Zea species. Sequencing of microsatellite alleles revealed complex patterns of mutation including frequent indels in the regions flanking microsatellite repeats. In one case, all variation at a microsatellite locus came from indels in the flanking region rather than in the repeat motif. Maize microsatellites show great variability within populations and provide a reliable means to measure intraspecific variation. Phylogeographic relationships of Zea populations were successfully reconstructed with good resolution using a genetic distance based on the infinite allele model, indicating that microsatellite loci are useful in evolutionary studies in Zea. Microsatellite loci show a principal division between tropical and temperate inbred lines, and group inbreds within these two broad germplasm groups in a manner that is largely consistent with their known pedigrees. Received: 10 February 2001 / Accepted: 21 May 2001     

Dopaminergic modulation of visual responses in toadsI. Apomorphine-induced effects on visually directed appetitive and consummatory prey-catching behavior

This study confirms for a phylogenetically basal terrestrial vertebrate that dopaminergic modulations interfere with the visually directed appetitive and consummatory feeding behaviors orienting and snapping, respectively. (1) In common toads Bufo bufo, intralymphatic administration of the dopamine D₂/D₁-receptor agonist apomorphine led to a dose-dependent facilitation of prey-snapping in response to moving objects. The snapping activity reached a maximum 15–35 min after apomorphine injection. (2) To changes in configurational stimulus features, the basic pattern of discrimination was maintained; however, the acuity of discrimination was reduced due to the high snapping response level. (3) The apomorphine-induced facilitation of snapping was accompanied by a suppression of prey-oriented lunging and turning. Toads snapped only if prey occurred frontally in the visual field at a relatively short distance. The snapping behavior was fixed in its form and stereotyped regarding its immediate release. (4) About 90 min after apomorphine administration, prey-oriented turning behavior was restored and displayed a facilitatory rebound. (5) In comparative experiments with the species B. marinus, both prey-oriented turning and snapping responses were suppressed by apomorphine in a dose-dependent manner. (6) After pre-treatment with the dopamine antagonist haloperidol, apomorphine showed no measurable effect on the visual release of prey orienting or snapping. (7) The results contribute to the sensorimotor and the motivation hypothesis of dopamine function proposed for higher vertebrates and stimulate a comparative discussion of anatomic homologies and functional analogies. Accepted: 10 July 1996     

EXAFS studies on the PN and POX states of the P-clusters in nitrogenase

The MoFe protein of nitrogenase is an α₂β₂ tetramer that contains two each of two different types of metal centers, the FeMo-cofactor and the P-clusters. The function of the P-clusters is believed to be to accept electrons from the Fe protein of nitrogenase and to donate them to the FeMo-cofactor. We have studied the P-clusters of Azotobacter vinelandii nitrogenase in both the P^N and P^OX states utilizing Fe K-edge X-ray absorption spectroscopy. Since the MoFe holoprotein contains the seven iron FeMo-cofactor centers in addition to P-clusters, we have utilized a FeMo-cofactor-deficient MoFe protein synthesized by the Δnif H strain DJ54. That MoFe protein is an α₂β₂ tetramer that contains P-clusters by the criteria of metal analysis, CD spectroscopy, cluster extrusion, and electrochemical reduction of the P^OX state. Several important results have emerged from our XAS studies. The first shell Fe-S coordination shows the same average Fe-S distance (2.26?Å) in both states. The second coordination shells could only be well fit using two different Fe-Fe contributions. In both states, short Fe-Fe components with distances of 2.57?Å and 2.42?Å for the P^N and P^OX states, respectively, were required to complement longer 2.75?Å and 2.70?Å distances. Understanding of the P-cluster structure is essential if we are to make advances in understanding the role of the P-clusters and their participation in electron transfer through the nitrogenase system.     

Some simple relationships between land-surface emissivity, greenness and the plant cover fraction for use in satellite remote sensing

 Simple equations are given which describe the relationships between the land-surface emissivity, the normalized difference vegetation index and the fractional vegetation cover. The empirical equations are validated using data taken from the literature. Adequate agreement is found between the formulas used in this study and those proposed by other authors. Received: 9 December 1996 / Revised: 23 May 1997 / Accepted: 4 July 1997     

Mapping of the Rf-3 nuclear fertility-restoring gene for WA cytoplasmic male sterility in rice using RAPD and RFLP markers

The cytoplasmic male sterility (CMS) of wild-abortive (WA) cytoplasm has been widely used for breeding hybrid rice. Two restorer genes for the CMS have been found by traditional genetic analysis. To tag the restorer genes we used a set of near-isogenic lines (NILs) of Zhenshan 97 carrying different genotypes for fertility restoration from IR24, to perform RAPD analysis. From the survey of 720 random primers, six RAPD markers were identified to be associated with Rf-3. Three of these OPK05-800, OPU10-1100 and OPW01-350, were mapped on chromosome 1. Two populations from the crosses between Zhenshan 97 A and a near-isogenic restorer line ZSR21 and between Zhenshan 97 A and IR24 were used for mapping Rf-3. The three RAPD markers and three RFLP markers, RG532, RG140 and RG458, were found to be closely linked to Rf-3 in the two populations. The same location of Rf-3 was also found in a population from the cross of IR58025 A//IR36/IR58025 B. At the RG532 locus, different alleles were found between two CMS lines, Zhenshan 97 A and IR58025 A, and between two restorer lines, IR24 and IR36. The use of these molecular markers closely linked to Rf-3 in facilitating the development of hybrid rice is discussed. Received: 3 January 1996 / Accepted: 17 May 1996     

Natural and synthesized ectomycorrhizas of the alpine dwarf willow Salix herbacea

 A new approach for selecting sampling sites of ectomycorrhizal roots is presented in order to describe ectomycorrhizas of Salix herbacea. Based on sporocarp mapping and statistical evaluation of the mapping data, sites for ectomycorrhizal root sampling were chosen underneath sporocarps of ectomycorrhizal Cortinarius (Myxacium) favrei, Hebeloma repandum, Laccaria montana, Entoloma alpicola, and Russula norvegica. Only in the samples beneath C. favrei, E. alpicola, and L. montana were corresponding ectomycorrhizas predominant and therefore described. Cenococcum geophilum ectomycorrhizas occurred throughout most samples and were also described. Numerous carpophores of the five selected ectomycorrhizal fungi were sampled for isolation purposes. Pure cultures were obtained of H. repandum and C. favrei, but laboratory syntheses of ectomycorrhizas were successful only with H. repandum and seedlings or cuttings of S. herbacea. Accepted: 27 February 1996     

Levels of genetic diversity at different stages of the domestication cycle of interior spruce in British Columbia

Concerns over the reductionist nature of the domestication of forest-tree species focus on the possibility of potential genetic erosion during this process. To address these concerns, genetic diversity assessments in a breeding zone the Province of British Columbia “interior” spruce (Picea glauca×engelmanni) program was conducted using allozyme markers. Genetic-variation comparisons were made between natural and production (seed orchard) populations as well as seed and seedling crops produced from the same breeding zone’s seed orchard. The natural population sample consisted of a total of 360 trees representing three stands within each of three watersheds present in the Shuswap-Adams low-elevation zone of interior British Columbia. Small amounts of genetic differentiation were observed among the nine natural populations (4%) and this was attributable to extensive gene flow Consequently, the sum of these nine populations was considered as a baseline for the genetic variation present in the breeding zone. The comparisons between the seed orchard and the breeding zone produced a similar percentage of polymorphic loci while the expected hetrozygosity (0.207 vs 0.210) and the average number of alleles per locus (2.7 vs 2.4) were slightly lower in the seed orchard. A total of seven natural populations’ rare alleles were not present in the orchard population, while one allele was unique to the orchard. The %P increased to 70.6% in the seedlot, but dropped to the natural populations level (64.7%) in the plantation. The observed increase in %P was a result of pollen contamination in the orchard. It is suspected that the reduction in the plantation was caused by an unintentional selection in the nursery. Simulated roguing in the orchard did not drastically reduce even if up to 50% of the orchard’s clones were rogued. However, roguing was associated with a reduction in the average number of alleles per locus (i.e., sampling effect). Received: 2 January 1996 / Accepted: 24 May 1996     

Plastid position in Arabidopsis columella cells is similar in microgravity and on a random-positioning machine

 In order to study gravity effects on plant structure and function, it may become necessary to remove the g-stimulus. On Earth, various instruments such as clinostats have been used by biologists in an attempt to neutralize the effects of gravity. In this study, the position of amyloplasts was assayed in columella cells in the roots of Arabidopsisthaliana (L.) Heynh. seedlings grown in the following conditions: on Earth, on a two-dimensional clinostat at 1 rpm, on a three-dimensional clinostat (also called a random-positioning machine, or an RPM), and in space (true microgravity). In addition, the effects of these gravity treatments on columella cell area and plastid area also were measured. In terms of the parameters measured, only amyloplast position was affected by the gravity treatments. Plastid position was not significantly different between spaceflight and RPM conditions but was significantly different between spaceflight and the classical two-dimensional clinostat treatments. Flanking columella cells showed a greater susceptibility to changes in gravity compared to the central columella cells. In addition, columella cells of seedlings that were grown on the RPM did not exhibit deleterious effects in terms of their ultrastructure as has been reported previously for seedlings grown on a two-dimensional clinostat. This study supports the hypothesis that the RPM provides a useful simulation of weightlessness. Received: 5 January 2000 / Accepted: 22 February 2000     

Photochemically driven models of oxygenases based on the use of iron porphyrins

Photoexcited iron porphyrins can be used to mimic the catalytic activity of cytochrome P-450 oxygenases both in the reduction of halogenated alkanes and in the oxidation of hydrocarbons by O₂ itself at room temperature and atmospheric pressure. The results reported indicate that there is a similarity and a complementarity of photonic activation with other more conventional methods of activation of model systems of oxygenases. In fact, by irradiating at suitable wavelengths, it is possible to induce redox reactions which mimic those of natural oxygenases, avoiding the difficulties caused by parallel processes which could be expected when chemical reagents are used. These processes occur with good turnover values of the photocatalyst and in mild temperature and pressure conditions. By controlling the reaction environment, it is possible to address the selectivity of the process. In this regard, the very recent results obtained after heterogenization of the photocatalysts inside membranes of Nafion® are particularly promising for the development of new biomimetic photocatalysts in heterogeneous or organized systems.     

Genetic diversity and classification of cyanobacteria in different Azolla species by the use of PCR fingerprinting

Symbiotically associated cyanobacteria from 18 accessions within all known species in the genus Azolla were examined and classified by the use of polymerase chain reaction (PCR)-fingerprinting. A repetitive sequence specific for cyanobacteria, the short tandemly repeated repetitive (STRR) sequence, was used as a primer in the reaction. Cyanobacterial filaments isolated directly from the Azolla leaf cavity or contained within homogenised symbiotic Azolla tissue were used as templates. Based on the fingerprint pattern, distinct differences were demonstrated between cyanobacteria isolated from the Euazolla and Rhizosperma sections. In addition, individual fingerprints were obtained from all cyanobacteria isolated from the different Azolla species. The fingerprints were used to generate a phylogenetic tree. Three clusters were distinguished: one contained the four isolates from the section Euazolla, a second the isolate from Azolla filiculoides, and a third the three isolates from the section Rhizosperma. By the use of STRR-PCR fingerprinting, new data on the taxonomy of cyanobacteria in Azolla were obtained, which have been difficult to generate by other classification methods. PCR-fingerprinting may, therefore, be a valuable tool for diversity and classification studies of symbiotic cyanobateria from Azolla and, as co-evolution between the cyanobacteria and its corresponding host exists the method may also be useful for the taxonomy of Azolla. Received: 19 December 1998 / Accepted: 31 May 1999     

Catabolism of native and oxidized low density lipoproteins: in vivo insights from small animal positron emission tomography studies

Summary. The human organism is exposed to numerous processes that generate reactive oxygen species (ROS). ROS may directly or indirectly cause oxidative modification and damage of proteins. Protein oxidation is regarded as a crucial event in the pathogenesis of various diseases ranging from rheumatoid arthritis to Alzheimer’s disease and atherosclerosis. As a representative example, oxidation of low density lipoprotein (LDL) is regarded as a crucial event in atherogenesis. Data concerning the role of circulating oxidized LDL (oxLDL) in the development and outcome of diseases are scarce. One reason for this is the shortage of methods for direct assessment of the metabolic fate of circulating oxLDL in vivo. We present an improved methodology based on the radiolabelling of apoB-100 of native LDL (nLDL) and oxLDL, respectively, with the positron emitter fluorine-18 (¹⁸F) by conjugation with N-succinimidyl-4-[¹⁸F]fluorobenzoate ([¹⁸F]SFB). Radiolabelling of both nLDL and oxLDL using [¹⁸F]SFB causes neither additional oxidative structural modifications of LDL lipids and proteins nor alteration of their biological activity and functionality, respectively, in vitro. The method was further evaluated with respect to the radiopharmacological properties of both [¹⁸F]fluorobenzoylated nLDL and oxLDL by biodistribution studies in male Wistar rats. The metabolic fate of [¹⁸F]fluorobenzoylated nLDL and oxLDL in rats in vivo was further delineated by dynamic positron emission tomography (PET) using a dedicated small animal tomograph (spatial resolution of 2 mm). From this study we conclude that the use of [¹⁸F]FB-labelled LDL particles is an attractive alternative to, e.g., LDL iodination methods, and is of value to characterize and to discriminate the kinetics and the metabolic fate of nLDL and oxLDL in small animals in vivo.     

An appraisal of methods used in coral recruitment studies

 A new method for attaching individual artificial settlement plates directly to the reef surface using small stainless steel base plates is described. Recruitment of corals to settlement plates attached to the reef substratum and to steel mesh racks is compared. The effects of differences in depth, settlement plate angle, and local topography on recruitment of corals were also investigated. No significant difference in mean recruit density was found between settlement plates deployed using the two attachment methods. Small differences in depth and plate angle among replicate plates explained less than 6% of the variability in coral recruitment on replicate settlement plates. The direct-attachment method is less obtrusive, more cost and time efficient, and settlement plates can be deployed at precise locations. Additionally, because settlement plates are deployed individually rather than grouped on racks or frames, the direct-attachment method avoids complications associated with assumptions of independence implicit in most statistical procedures. Accepted: 24 November 1999