Plasticity of the landing response of Drosophila melanogaster

Summary Fruitflies (Drosophila melanogaster) may respond with landing reactions in tethered flight to unilateral progressive motion of single vertical dark stripes. The response frequency to repeated unilateral stimulation has a characteristic time course, a fast increase followed by a slower decrease. This behaviour is explained by the habituation of the input channels to a leaky integrator. The half-life of the integrated signal is in the range of 1 s. Contralateral sensitization (CoS) describes the sensitization of the landing response to unilateral stimuli by preceding contralateral stimulation. It acts by increasing the initial reactivity, which habituates. The effects of CoS are thus still obvious after 1 min of repetitive stimulation. CoS can effectively be mediated by movement stimuli as well as by flickering light. We also show that binocular rotation rather than unilateral back to front motion inhibits the landing response (in the monocular part of the visual field). The biological significance of the described temporal characteristics of the landing response system and their possible neuronal basis are discussed.Abbreviations CoS contralateral sensitization     

Anti-predator strategies in oviposition site selection of Pyrrhosoma nymphula (Zygoptera: Odonata)

Summary Selection of sites for oviposition when the risk of predation by green frogs was variable studied in tandems of the damselfly Pyrrhosoma nymphula. In the absence of predators the number of tandems landing increased with the size of the oviposition site and with the number of pairs already present. Pairs approaching an oviposition site landed promptly and preferred locations near other ovipositing pairs. Pairs which had landed near others left the landing site and flew to another water plant after a shorter period than those landing alone. They stayed in groups for only 16.5% of the total duration of stay at the oviposition site. With predators present fewer tandems landed. There was no relationship between the landing site of solitary tandems and the locations of the perching frogs. Pairs landing in groups were disturbed by frogs after a shorter period than pairs landing alone. Damselfly aggregation did not affect the predation success of the frogs, but the predation risk to individual tandem pairs was reduced.     

Comparison between the movement detection systems underlying the optomotor and the landing response in the housefly

Flies evaluate movement within their visual field in order to control the course of flight and to elicit landing manoeuvres. Although the motor output of the two types of responses is quite different, both systems can be compared with respect to the underlying movement detection systems. For a quantitative comparison, both responses were measured during tethered flight under identical conditions. The stimulus was a sinusoidal periodic pattern of vertical stripes presented bilaterally in the fronto-lateral eye region of the fly. To release the landing response, the pattern was moved on either side from front to back. The latency of the response depends on the stimulus conditions and was measured by means of an infrared light-beam that was interrupted whenever the fly lifted its forelegs to assume a preprogrammed landing posture (Borst and Bahde 1986). As an optomotor stimulus the pattern moved on one side from front to back and on the other side in the opposite direction. The induced turning tendency was measured by a torque meter (Götz 1964). The response values which will be compared are the inverse latencies of the landing response and the amplitude of the yaw torque.

1. Optomotor course-control is more sensitive to pattern movement at small spatial wavelengths (10° and 20°) than the landing response (Fig. 1a and b). This suggests that elementary movement detectors (EMDs, Buchner 1976) with large detection base (the distance between interacting visual elements) contribute more strongly to the landing than to the optomotor system.
2. The optimum contrast frequencies of the different responses obtained at a comparatively high pattern contrast of about 0.6 was found to be between 1 and 10 Hz for the optomotor response, and around 20 Hz for the landing response (Fig. 2a and b). This discrepancy can be explained by the fact that the optomotor response was tested under stationary conditions (several seconds of stimulation) while for the landing response transient response characteristics of the movement detectors have to be taken into account (landing occurs under these conditions within less than 100 ms after onset of the movement stimulus). To test the landing system under more stationary conditions, the pattern contrast had to be reduced to low values. This led to latencies of several seconds. Then the optimum of the landing response is around 4 Hz. This is in the optimum range of the optomotor course-control response. The result suggests the same filter time constants for the movement detectors of both systems.
3. The dependence of both responses on the position and the size of the pattern was examined. The landing response has its optimum sensitivity more ventrally than the optomotor response (Fig. 3a and b). Both response amplitudes increase with the size of the pattern in a similar progression (Fig. 3c and d).

In first approximation, the present results are compatible with the assumption of a common set of movement detectors for both the optomotor course-control and the landing system. Movement detectors with different sampling bases and at different positions in the visual field seem to contribute with different gain to both responses. Accordingly, the control systems underlying both behaviors are likely to be independent already at the level of spatial integration of the detector output.     

Bionomics of Anopheles spp. (Diptera: Culicidae) in a malaria endemic region of Sukabumi,West Java,Indonesia

A 15‐month bionomic study of Anopheles species was conducted in two ecologically distinct villages (coastal and upland) of Sukabumi District, West Java, Indonesia from June 2006 to September 2007. Mosquitoes were captured using human‐landing collections at both sites. During the study, a total of 17,100 Anopheles mosquitoes comprising 13 Anopheles species were caught: 9,151 at the coastal site and 7,949 at the upland site. Anopheles barbirostris, Anopheles maculatus, and Anopheles vagus were the predominant species caught at the coastal site, and Anopheles aconitus, Anopheles barbirostris, and An. maculatus predominated in the upland site. Overall, species were exophagic at both sites, but there was variation between species. Anopheles aconitus was endophagic at the coastal site, exophagic at the upland site, collected most often in April 2007 and had a peak landing time between 22:00 and 23:00. Anopheles sundaicus was only collected at the coastal site, exophagic, collected most often in October 2006, and had a peak landing time between 19:00 and 20:00. Potential malaria vector species such An. aconitus, An. maculatus, and An. sundaicus were present throughout the year. None of the 7,770 Anopheles tested using CSP‐ELISA were positive for malaria, although the risk for malaria outbreaks in Sukabumi district remains high.     

What kind of movement detector is triggering the landing response of the housefly?

As shown before, the latency of the housefly's landing response depends on the conditions of the visual stimulus (Borst 1986). Accordingly, the latency can be used to characterize the movement detection system which is triggering the landing response.The stimulus was a sinusoidal periodic pattern of vertical stripes presented bilaterally in the frontolateral eye region of the fly. It started to move, simultaneously on either side, from front to back at a given time. The latency of the response was measured by means of an infrared light-beam that was interrupted whenever the fly lifted its forelegs to assume a preprogrammed landing posture (Fig. 1). The latency was found to vary in a range from 60 ms up to several seconds depending on the pattern's spatial wavelength , contrast frequency cf and contrast C.For sufficiently high pattern contrast the optimum of the reaction (minimum latency) is found at spatial wavelengths of 30–40° and contrast frequencies of 8–17 periods/s (Fig. 3a). This is about 2–10 times more than is anticipated from the optomotor response under similar conditions. Evaluation of the optimum contrast frequency cf _OPT at different wavelengths shows that cf _OPT is not independent of (Fig. 3b, solid line). The same is true for the contrast dependence of the reaction: reduction of the contrast leads not only to a general decrease in the response amplitudes (prolongation of the latency) (Fig. 4a), but also to a shift of cf _OPT towards lower contrast frequencies (Fig. 4b, solid line).In the theory of the correlation-type movement detector (Reichardt 1961) which underlies the optomotor response of flies the dependence of cf _OPT on pattern wavelength and/or pattern contrast is not expected under stationary conditions. However, as shown by computer simulation all experimental results can be explained by a homogeneous retinotopic array of correlation movement detectors (Fig. 2) if their response under non-stationary conditions is taken into account. We simply assume that the spatially and temporally integrated output of the movement detectors is evaluated by a threshold device (Fig.5). The correlation-type movement detection in combination with a temporal integrator system predicts the rather complex dependence of the optimum contrast frequency on pattern wavelength and pattern contrast (dashed lines in Fig. 3b and 4b) and provides the missing explanation of the variable latencies of the landing response.Comparing the parameters of the correlation-type movement detector derived in the present study with those of the optomotor response, the landing response seems to use the same type of movement detection system. To account for the high wavelength optimum, however, the input elements of the movement detection system of the landing response might have an increased visual field (e.g. by pooling neighbouring visual elements) and, accordingly, a reduced visual acuity as compared with the input elements of the optomotor system.Abbreviations (°) spatial pattern wavelength - w(°/s) angular velocity of the pattern - cf (Hz) contrast frequency=w/ - cf _OPT(Hz) cf leading to the shortest latency - (Hz) angular frequency=2cf - I mean luminance of the pattern - I modulation amplitude of the pattern - C pattern contrast=I/ - (ms) time constant of a filter - (°) angle between the optical axis of neighbouring visual elements - (°) acceptance angle of visual elements     

Effect of landing height on frontal plane kinematics, kinetics and energy dissipation at lower extremity joints

Lack of the necessary magnitude of energy dissipation by lower extremity joint muscles may be implicated in elevated impact stresses present during landing from greater heights. These increased stresses are experienced by supporting tissues like cartilage, ligaments and bones, thus aggravating injury risk. This study sought to investigate frontal plane kinematics, kinetics and energetics of lower extremity joints during landing from different heights. Eighteen male recreational athletes were instructed to perform drop-landing tasks from 0.3- to 0.6-m heights. Force plates and motion-capture system were used to capture ground reaction force and kinematics data, respectively. Joint moment was calculated using inverse dynamics. Joint power was computed as a product of joint moment and angular velocity. Work was defined as joint power integrated over time. Hip and knee joints delivered significantly greater joint power and eccentric work (p<0.05) than the ankle joint at both landing heights. Substantial increase (p<0.05) in eccentric work was noted at the hip joint in response to increasing landing height. Knee and hip joints acted as key contributors to total energy dissipation in the frontal plane with increase in peak ground reaction force (GRF). The hip joint was the top contributor to energy absorption, which indicated a hip-dominant strategy in the frontal plane in response to peak GRF during landing. Future studies should investigate joint motions that can maximize energy dissipation or reduce the need for energy dissipation in the frontal plane at the various joints, and to evaluate their effects on the attenuation of lower extremity injury risk during landing.     

Orientation sensitivity of the visual movement detection system activating the landing response of the blowflies,Calliphora, and Phaenicia: A behavioural investigation

The orientation sensitivity of the visual movement detection system relative to the axes of the eye was investigated for the landing response by changing the direction of movement of a periodic striped pattern (unidirectional movement) and a two-stripe pattern consiting of two stripes moving apart (bidirectional movement). In the momocular, equatorial regions of the eye progressive motion proves to be most effective, whereas in the frontal (equational), binocular region descendive motion is most effective in eliciting the landing response, probably caused by binocular interactions. A strong enhancement of the response is induced by stimulation in the binocular region of the two eyes. The orientation of elementary movement detectors relative to the axes of the ommatidial array is discussed. The findings are summarized in a functional model of the landing response.     

Initial preference for plant species and state during oviposition site selection by an odonate

Endophytic egg‐laying odonates use an ovipositor to insert their eggs inside plant tissues. Before egg deposition, oviposition site selection consists of two crucial steps: (i) the initial choice, typically decided in species that oviposit in tandem within vertical substrates by the male when landing and then by the female by staying on the substrate or flying to another; and (ii) the insertion site choice, made by the female who uses her ovipositor to palpate the substrate. Some odonates prefer to deposit their eggs within specific plant species. Some are able to discriminate between living and dead substrates during the initial choice. However, the extent to which odonates discriminate among distinct plant species during the initial choice is unknown. We studied the initial site preference in Lestes macrostigma (Odonata: Lestidae) to determine whether the males and/or females show a distinct preference among five types of shoots when landing on or when palpating the substrate, respectively. Male L. macrostigma preferred to land on Bolboschoenus maritimus and dead Juncus spp. When focusing on J. maritimus, females preferentially palpated the substrate when the male landed on dead shoots. We suggest that the male preference for these substrates is consistent with that of the female during insertion site choice but also during egg deposition. Such behavior should reduce the duration of oviposition, with benefits of reducing the predation risk and increasing available time for foraging. The advantage in preferring these substrates should be linked to a selection pressure acting on egg development and/or survival.     

The Mode of Orientation during Flight and Approach to Landing in two Phyllostomid Bats

The use of vision during flight and approach to a landing site in two phyllostomid bat species, Phyllostomus discolor and Desmodus rotundus was investigated. Three individuals of each species were trained to traverse a 3-m flight tunnel and to land at a small illuminated grid that was randomly changed between two positions on a front wall. Analysis of the flight path by observation and different technical methods revealed that the bats oriented themselves toward the grid quite early. When the illumination was switched off the flight path diverged much later. With a dark landing grid on one side and an optical projection of it on the other the bats aimed towards this dummy, interrupted the approach 20–40 cm in front of the illusion and then tried to reach the other side or turned back. Whether microchiroptera may rely in the medium range more on vision than usually is thought is discussed.     

Unusual Behavior of Polygyne Fire Ant Queens on Nuptial Flights

This study reports previously undescribed behavior of fire ant queens (Solenopsis invicta) on their nuptial flights. We captured large numbers of alate (winged) queens flying at low altitudes in dense swarms that were virtually devoid of males. We assayed the genotypes of these alate queens at the locus Gp-9, which exhibits strong genotype frequency differences between monogyne (single-queen) and polygyne (multiple-queen) populations, and found that almost all of these low-flying queens originated from polygyne colonies. Comparisons of mtDNA haplotype distributions of these queens to those of alates leaving polygyne nests suggest that the flying queens had not dispersed more than a few hundred meters. Moreover, the proportion of flying queens that were mated did not differ significantly from the proportion of reproductive queens that were mated within the same sites. Thus the flight behavior appears to occur subsequent to mating. We suggest that the flying queens are sampling the local environment in order to select a suitable landing site. Such a site would contain established polygyne nests into which the queens may be adopted as new reproductives.     

Aspects of maturation and reproduction in hexanchiform and squaliform sharks

Three species of hexanchiform sharks belonging to two families and 12 species of squaliform sharks belonging to three families were recorded in fish landing site surveys in eastern Indonesia. Of these, the Squalidae were the most abundant species landed, with Squalus hemipinnis, Squalus edmundsi and Squalus montalbani contributing 0·4, 0·4 and 0·5% to the total number of sharks recorded in a 5 year survey of Indonesian fish landing sites. In comparison, the hexanchid Hexanchus griseus contributed the largest percentage to the total shark biomass. For many species, the majority of the catch consisted of immature fishes, which had not yet been able to reproduce. The data presented in this article are the first biological data reported on most of these shark species and are thus vital for fisheries managers and conservation assessors.     

Aggregates of Resident Bacteria Facilitate Survival of Immigrant Bacteria on Leaf Surfaces

The fate of immigrant bacterial cells on leaves under stressful conditions was determined as a function of the anatomical features and the local spatial density of resident cells at their landing site. Pantoea agglomerans 299R was established on bean leaves and the survival of immigrant cells of Pseudomonas fluorescens A506 and Pseudomonas syringae B728a, as well as P. agglomerans itself, was determined by epifluorescence microscopy following subsequent exposure of plants to desiccation stress. Resident and immigrant bacterial strains constitutively expressed the cyan and the green fluorescent protein, respectively, and the viability of individual cells was assessed directly on leaf surfaces following propidium iodide staining. Although only a small fraction of the immigrant cells landed on established bacterial aggregates, their fate was usually strongly influenced by the presence of indigenous bacteria at the site at which they landed. Immigrants of P. agglomerans 299R or P. fluorescens A506 that arrived as solitary cells had about double the probability of survival when landing on aggregates formed by P. agglomerans 299R than when landing on uncolonized areas of the leaf surface. In contrast, the survival of P. syringae B728a was similar irrespective of whether it landed on colonized or uncolonized parts of a leaf. The nature of plant anatomical features at which immigrant bacteria landed also strongly influenced the fate of immigrant bacteria. The fraction of immigrant cells of each species tested that landed on veins, glandular trichomes, or epidermal cells altered by P. agglomerans that died was always less than when they landed on normal epidermal cells or at the base of hooked trichomes. Depending on the process by which immigrants arrive at a leaf, only a small fraction of cells may be deposited on existing bacterial aggregates. Although uncolonized sites differed greatly in their ability to influence the survival of immigrant cells, the fate of an immigrant bacterium will depend on the nature of the leaf structure on which it is deposited, and apparently indirectly on the amount of nutrients and water available at that site to support the development of bacterial aggregates.     

The Differences in Lower Extremity Joints Energy Dissipation Strategy during Landing between Athletes with Symptomatic Patellar Tendinopathy (PT) and without Patellar Tendinopathy (UPT)

Patellar tendinopathy is a clinical symptom of patellar tendons characterized by local pain in the front of the knee joint. It is common among basketball and volleyball players. Patients with patellar tendinopathy may exhibit different landing strategies during landing compared to healthy individuals. The purpose of this study was to compare the differences in lower limb joint energy dissipation (eccentric work) values for the symptomatic patellar tendinopathy (PT) athletes and no patellar tendinopathy (UPT) athletes during single-leg landing. A total of 26 (PT: 13, UPT:13) semi-professional male basketball and volleyball player’s kinetic data were collected during the landing phases. Joint work was calculated by the integral of joint power over time. In this study, the result showed that the ankle joint means energy dissipation (p < 0.001) and total energy dissipation (p < 0.001) of PT were significantly greater than UPT. Compared to the UPT athletes, the PT athletes showed smaller knee joint mean energy dissipation (p = 0.002) and contribution to total energy dissipation (p < 0.001) during the landing stance. Meanwhile, there were no differences in hip joint contribution to total energy dissipation (p = 0.523) and lower limb total energy dissipation (p = 0.127). A deeper understanding of each joint’s energy dissipation contribution ratio between UPT and PT during landing can provide clues that reveal the biomechanical mechanism of PT athletes landing. Further study should choose a larger sample size to more comprehensively reveal the energy dissipation strategy of PT during landing.     

A subsequent movement alters lower extremity muscle activity and kinetics in drop jumps vs. drop landings

Drop landings and drop jumps are common training exercises and injury research model tasks. Drop landings have a single landing, whereas drop jumps include a subsequent jump after initial landing. With the expected ground impact, instant and landing surface suggested to modulate landing neuromechanics, muscle activity, and kinetics should be the same in both tasks when landing from the same height onto the same surface. Although previous researchers have noted some differences between these tasks across separate studies, little research has compared these tasks in the same study. Thus, we examined whether a subsequent movement after initial landing alters muscle activity and kinetics between drop landings and jumps. Fifteen women performed 10 drop landings and drop jumps each from 45 cm. Muscle onsets and integrated muscle activation amplitudes 150 milliseconds before (preactivity) and after landing (postactivity) in the medial and lateral quadriceps, hamstrings, and lateral gastrocnemius and peak and time-to-peak vertical ground reaction forces were examined across tasks (p ≤ 0.05). When performing drop jumps, subjects demonstrated later (p = 0.02) gastrocnemius and lesser lateral gastrocnemius (p = 0.002) and medial quadriceps (p = 0.02) preactivity followed by increased postactivity in all muscles (p = 0.006), with higher peak vertical ground reaction forces (p = 0.04) but no differences in times to these peaks (p = 0.60) than drop landings. The later gastrocnemius activation, higher gastrocnemius and quadriceps postlanding amplitudes, and higher ground reaction forces in drop jumps may allow subjects to propel the body vertically after the initial landing vs. simply absorbing impact in drop landings. Our results indicate that in addition to landing surface and height, anticipation of a subsequent task changes landing neuromechanics. Generalizations of results from landing-only studies should not be made with landing followed-by-subsequent-activity studies. Landing exercises should be incorporated based on sport-specific demands.     

Analysis of secondary, integration sites for IS117 in Streptomyces lividans and their role in the generation of chromosomal deletions

IS117, the 2.6 kb mini-circle of Streptomyces coelicolor A3(2), is a transposable element previously shown to be integrated into two distant sites in the chromosome. When introduced into S. lividans, IS117 integrates into one preferred chromosomal site, but when this site was artificially deleted, IS117 integrated into many secondary sites. Nucleotide sequence analysis of several secondary integration sites revealed varying degrees of similarity with the preferred site, but no consensus sequence. Nevertheless, sites more similar to the preferred site tended to be occupied more often than those that are less similar. Insertion of IS117 into secondary sites in the chromosome of S. lividans sometimes mediated chromosomal rearrangements. It was shown that some strains containing IS117 integrated into secondary sites had suffered deletions of chromosomal DNA. Deletions were adjacent to the inserted element and were at least several kilobases long. The proposed model implicates homologous recombination between IS117 copies integrated into two different secondary sites in the same chromosome as a cause of the deletions.     

Hopping and swimming in the leopard frog,Rana pipiens: I. Step cycles and kinematics

This study presents a model for the step cycle patterns used during both hopping and swimming by the leopard frog, Rana pipiens. The two behaviors are essentially similar in movement pattern and in the ways they are modified from quadrupedal gaits. In hopping, there is marked hind limb extension throughout stance. The swing begins with a suspension equivalent to the leap that occurs in a galloping or bounding quadruped. Following suspension, as the frog descends from the apex of its leap, the hind limbs remain posterior and in line with the spine while they flex. Near the end of flexion, there is a rapid downward rotation of the hindquarters to bring the hind feet underneath the body. This movement utilizes the planted forelimb as a pivot. A similar pattern of movement occurs in swimming; the stance (propulsion) phase involves extension at all hind limb joints. The swing (recovery) phase begins with the hind feet fully extended and includes a protracted gliding phase, equivalent to the suspension in the hop. The hind limb then recovers to its initial position during a flexion phase. Since there is no landing and the hind limbs remain lateral rather than ventral to the pelvis, less flexion occurs in the spine or the limb joints. In both behaviors, the extensor muscles of hip (M. semimembranosus), knee (M. cruralis), and ankle (M. plantaris longus) achieve their longest lengths, when they likely can produce near maximal force, at the beginning of extension. All three muscles shorten during extension, but, because they are multiple-joint muscles, the amount of shortening is relatively small (≈ 15%). Hopping and swimming in frogs are comparable asymmetrical gaits with the same relative contact intervals (25% of stride). The step cycles in both gaits are modified from quadrupedal locomotion in the same ways: by 1) loss of knee and ankle extension toward the ground prior to landing (or end of flexion in swimming), 2) loss of a yield phase on landing (or end of flexion in swimming), and 3) inclusion of extended suspensions in both gaits. © 1996 Wiley-Liss, Inc.     

Catch composition and aspects of the biology of sharks caught by Thai commercial fisheries in the Andaman Sea

Catch composition, landing patterns and biological aspects of sharks caught by commercial fishing fleet operating in the Andaman Sea were recorded from landing sites in Ranong province of Thailand over a period of 1 year. Of the 64 species previously reported in the existing Thailand checklist, only 17 species were recorded in this study. Shark landings from the Andaman Sea appear now to be dominated largely by bamboo sharks Chiloscyllium spp. (Hemiscylliidae), which contribute c. 65% of the total number of sharks recorded. The carcharhinid sharks comprised c. 30·5% to the total catch, while the remaining c. 4·5% of landings comprised sharks from the families Squalidae, Stegostomatidae, Sphyrnidae and Triakidae. The catch composition is remarkably different from the previous landing survey in 2004, in that the current study found noticeable declines in landings of slow‐growing, late‐ maturing and low‐fecundity species (especially sphyrnid and carcharhinid species). The absences of many species and changes in life‐stage composition suggest that the populations of these groups may be close to collapse. The results from this study emphasize the urgency for additional research and monitoring efforts and also the need for management incentives in order to manage shark fisheries effectively in the Andaman Sea.     

Movement of the Southern Australian abalone Haliotis laevigata in relation to crevice abundance

The long-term movement of the abalone Haliotis laevigata was measured at three sites in Waterloo Bay, South Australia, characterized by differing amounts of available crevice space. Movement was negligible at a site where crevices were abundant, but extensive at a site without crevice space. At a site of limited available crevice space, the amount of movement increased as available crevice space decreased. In addition, the extent of movement was size dependent and movement was oriented in the direction of the approaching swell. Current experimental designs for measuring natural mortality of abalone depend heavily on assumptions about their movement. An understanding of this behaviour is thus a prerequisite for designing experiments to measure the mortality rates of abalone or other sedentary animals. The natural mortality rate at one site was estimated to be 0.59 (s.e.m. = 0.02, i.e. survival = 56% per year) and the disappearance rate (natural mortality and emigration) was independent of sex.     

Effects of Barriers and Thermal Refugia on Local Movement of the Threatened Leopard Darter, Percina pantherina

Local, short-term dispersal by the U.S. federally-threatened leopard darter, Percina pantherina, was examined in the field and in the laboratory to assess the possible effects of natural versus man-made barriers on movement. Mark-resight studies were conducted in two summers at sites in the Glover River (southeastern Oklahoma, U.S.A.). At one site, patches of 'preferred' habitat were separated by a natural riffle; at the other site, by a low-water road crossing with culverts. At the Natural Riffle site, darters moved downstream across the riffle, but also moved upstream into deeper water when water temperatures exceeded 29°C in the `preferred' habitat. Use of deeper, cooler waters by this species in late summer suggests that thermal refugia may be important habitats for the long-term management of leopard darters. At the Road Crossing site, all documented movement was in a downstream direction, and at least two darters traversed culverts in the low-water bridge. Laboratory studies of movement across several types of culverts suggested that culverts significantly decrease the probability of movement among habitat patches.     

Generating Customized Transgene Landing Sites and Multi-Transgene Arrays in Drosophila Using phiC31 Integrase

Transgenesis in numerous eukaryotes has been facilitated by the use of site-specific integrases to stably insert transgenes at predefined genomic positions (landing sites). However, the utility of integrase-mediated transgenesis in any system is constrained by the limited number and variable expression properties of available landing sites. By exploiting the nonstandard recombination activity exhibited by a phiC31 integrase mutant, we developed a rapid and inexpensive method for isolating landing sites that exhibit desired expression properties. Additionally, we devised a simple technique for constructing arrays of transgenes at a single landing site, thereby extending the utility of previously characterized landing sites. Using the fruit fly Drosophila melanogaster, we demonstrate the feasibility of these approaches by isolating new landing sites optimized to express transgenes in the nervous system and by building fluorescent reporter arrays at several landing sites. Because these strategies require the activity of only a single exogenous protein, we anticipate that they will be portable to species such as nonmodel organisms, in which genetic manipulation is more challenging, expediting the development of genetic resources in these systems.