Phototactic Response Research Articles

Light Control of Localized Photobioconvection.

Microorganismal motility is often characterized by complex responses to environmental physico-chemical stimuli. Although the biological basis of these responses is often not well understood, their exploitation already promises novel avenues to directly control the motion of living active matter at both the individual and collective level. Here we leverage the phototactic ability of the model microalga Chlamydomonas reinhardtii to precisely control the timing and position of localized cell photoaccumulation, leading to the controlled development of isolated bioconvective plumes. This novel form of photobioconvection allows a precise, fast, and reconfigurable control of the spatiotemporal dynamics of the instability and the ensuing global recirculation, which can be activated and stopped in real time. A simple continuum model accounts for the phototactic response of the suspension and demonstrates how the spatiotemporal dynamics of the illumination field can be used as a simple external switch to produce efficient bio mixing.

Reentrant Efficiency of Phototaxis in Chlamydomonas reinhardtii Cells

Phototaxis is one of the most fundamental stimulus-response behaviors in biology wherein motile microorganisms sense light gradients to swim toward the light source. Apart from single-cell survival and growth, it plays a major role at the global scale of aquatic ecosystems and bioreactors. We study phototaxis of single-celled algae Chlamydomonas reinhardtii as a function of cell number density and light stimulus using high spatiotemporal video microscopy. Surprisingly, the phototactic efficiency has a minimum at a well-defined number density, for a given light gradient, above which the phototaxis behavior of a collection of cells can even exceed the performance obtainable from single isolated cells. We show that the origin of enhancement of performance above the critical concentration lies in the slowing down of the cells, which enables them to sense light more effectively. We also show that this steady-state phenomenology is well captured by modeling the phototactic response as a density-dependent torque acting on an active Brownian particle.

Short- and long-duration exposures to cell-phone radiofrequency waves produce dichotomous effects on phototactic response and circadian characteristics of locomotor activity rhythm in zebrafish, Danio rerio

ABSTRACT We examined the effects of short- and long-duration exposure to cell phone radiation (CPR) on phototactic response (PR) and circadian characteristics of locomotor-activity rhythm, respectively, in zebrafish. We exposed four groups (10 in each) randomly selected fish to CPR emitted by a 2300 MHz 4G cell phone for 0.5 h, 1 h, 2 h, 3 h, and 4 h duration at two different time points of a day, i.e. morning and evening, to assess the PR. We also studied the effects of 1 week CPR exposure on circadian rhythm in locomotor activity pattern in eight singly housed zebrafish. We had eight singly housed sham-exposed zebrafish as the control. Results of two-way ANOVA revealed statistically significant effects of the factors exposure time and exposure duration on the PR of zebrafish. Fish exhibited photo-positive response till 2 h following morning exposure, and thereafter, showed photo-negative response with increased duration of exposure. During evening exposure, the fish were mostly photo-negative. However, 1 week exposure to CPR did not produce any significant effects on the circadian characteristics of locomotor activity rhythm. It is concluded that the CPR might alter phototaxis, but not the parameters of circadian rhythm in locomotor activity pattern of zebrafish.

Advances in insect phototaxis and application to pest management: a review.

Many insects, especially nocturnal insects, exhibit positive phototaxis to artificial lights. Light traps are currently used to monitor and manage insect pest populations, and play a crucial role in physical pest control. Efficient use of light traps to attract target insect pests is an important topic in the application of integrated pest management (IPM). Phototactic responses of insects vary among species, light characteristics and the physiological status of the insects. In addition, light can cause several biological responses, including biochemical, physiological, molecular and fitness changes in insects. In this review, we discuss several hypotheses on insect phototaxis, factors affecting insect phototaxis, insect-sensitive wavelengths, biological responses of insects to light, and countermeasures for conserving beneficial insects and increasing the effect of trapping. In addition, we provide information on the different sensitivities to wavelengths causing positive phototactic behavior in > 70 insect pest and beneficial insect species. The use of advanced light traps equipped with superior light sources, such as light-emitting diodes (LEDs), will make physical pest control in IPM more efficient. © 2019 Society of Chemical Industry.

Analysis of the genetically tractable crustacean Parhyale hawaiensis reveals the organisation of a sensory system for low-resolution vision

BackgroundArthropod eyes have diversified during evolution to serve multiple needs, such as finding mates, hunting prey and navigating in complex surroundings under varying light conditions. This diversity is reflected in the optical apparatus, photoreceptors and neural circuits that underpin vision. Yet our ability to genetically manipulate the visual system to investigate its function is largely limited to a single species, the fruit fly Drosophila melanogaster. Here, we describe the visual system of Parhyale hawaiensis, an amphipod crustacean for which we have established tailored genetic tools.ResultsAdult Parhyale have apposition-type compound eyes made up of ~ 50 ommatidia. Each ommatidium contains four photoreceptor cells with large rhabdomeres (R1–4), expected to be sensitive to the polarisation of light, and one photoreceptor cell with a smaller rhabdomere (R5). The two types of photoreceptors express different opsins, belonging to families with distinct wavelength sensitivities. Using the cis-regulatory regions of opsin genes, we established transgenic reporters expressed in each photoreceptor cell type. Based on these reporters, we show that R1–4 and R5 photoreceptors extend axons to the first optic lobe neuropil, revealing striking differences compared with the photoreceptor projections found in related crustaceans and insects. Investigating visual function, we show that Parhyale have a positive phototactic response and are capable of adapting their eyes to different levels of light intensity.ConclusionsWe propose that the visual system of Parhyale serves low-resolution visual tasks, such as orientation and navigation, based on broad gradients of light intensity and polarisation. Optic lobe structure and photoreceptor projections point to significant divergence from the typical organisation found in other malacostracan crustaceans and insects, which could be associated with a shift to low-resolution vision. Our study provides the foundation for research in the visual system of this genetically tractable species.

Effects of sediment resuspension on the larval stage of the model sponge Carteriospongia foliascens

Sponges are important components of many marine communities and perform key functional roles. Little is known on the processes that drive larval dispersal and habitat selection in sponges, and in particular under stress scenarios. The increase in sediment in the marine environment is a growing concern for the health of ecosystems, but scarce information exists on the effects of sediment on sponge larvae. This study assessed the effects of suspended and deposited sediment on the larva of Carteriospongia foliascens. A suspended sediment concentration (SSC) of 100 mg L−1 caused homogenisation of the natural pattern of phototactic responses, leading to 100% of photonegative behaviours and a reduction of swim speeds by 27%. After 24 h exposure to suspended sediments, fine particles were found attached to larval cilia, causing abnormal swimming behaviours. Larvae did not have the ability to remove the attached sediment that led to a transformation of the larval body into a cocoon-like morphology and death. Mortality tripled from 3 mg L−1 (9%) to 300 mg L−1 (30%) and the relative SSC EC10 and EC50 values corresponded to 2.6 mg L−1 and 17.6 mg L−1 respectively. Survival, as determined by live swimming larvae, exceeded 50% even in the highest SSC of 300 mg L−1, however settlement success decreased by ~20%. Larvae were able to settle onto substrate having deposited sediment levels (DSLs) up to 3 mg cm−2 (~24%), but recorded a 25 × chance of dislodgement compared to settlers on substrate with DSL of 0.3 mg cm−2. Larvae avoided settling onto substrates with DSLs >10 mg cm−2 and preferentially settled onto alternative vertical substrate that were free of sediment. While C. foliascens larvae have some ability to survive and settle through conditions of elevated sediment, detrimental effects are also clear.

Influence of green light at night on Juvenile hormone in the oriental armyworm Mythimna separata (Lepidoptera: Noctuidae)

AbstractThe oriental armyworm Mythimna separata is an agricultural insect pest in Eastern Asia. Mythimna separata moths have a high phototactic response to green (520 nm) light. The biological characteristics of insects living under light of a specific wavelength at night can change and, accordingly, Juvenile hormone (JH) levels may be influenced by this light. The present study evaluates changes in the total JH levels at different developmental stages (larvae, pupae and adults) of M. separata reared under green light with different exposure periods at night (or dark period). The results show that, when the exposure time per day of the green light at night is extended, the JH levels in the final‐instar larvae (22 days) and older age pupae (8 days) are significantly reduced, and the JH levels in earlier age pupae (4 days) and adults (3, 6 and 9 days) are significantly increased, compared with groups not exposed to green light. Additionally, the JH level of male moths significantly differs from that of the female moths. We suggest that the JH level of M. separata insects could be regulated by the green light at night (or dark period). The findings of the present study will help to explain the relationship between the light environment and biological characteristics in nocturnal moths.

Reproductive strategy of the floating alga Ulva prolifera in blooms in the Yellow Sea based on a combination of zoid and chromosome analysis

Green algal blooms have occurred in the Yellow Sea for 13 consecutive years since 2007. However, little is known about the reproductive strategy of the dominant species Ulva prolifera in the field. In particular, it is not clear whether the floating Ulva species are sporophytes or gametophytes, and if their life history is sexual or asexual. In this study, the life history type was determined based on the size, phototactic response, and flagella number for the zoids in at least two successive generations. In addition, chromosome observations were conducted to distinguish the gametophytes and sporophytes in the floating Ulva species. The results showed that the floating Ulva species were all sporophytes with sexual reproductive patterns, thereby indicating that this Ulva species always maintains vegetative growth from April to June during the early stage of the blooms. In addition, we found that the chromosome numbers were 18 for the diploid sporophytes and nine for the haploid male and female gametophytes. These results provide useful information to help understand the explosive growth of these green algal blooms.

Phototactic behavior of native Daphnia in the presence of chemical cues from a non-native predator Bythotrephes.

Chemical cues are used by many taxa to communicate within and among species. Behavioral defenses induced by predator cues are a mechanism by which prey species resist or avoid predator attack. This study examined the egg bank of native Daphnia species in a lake that has been invaded by Bythotrephes longimanus, an invertebrate zooplanktivore native to northern-central Europe and Asia (initial invasion 1994, population boom in 2009). Daphnia resting eggs from both pre- and post-B. longimanus invasion lake sediments were hatched and established as isofemale clonal lines. Phototactic behavior (a proxy for vertical migration behavior) was assessed in the presence and absence of B. longimanus cue. This was done to evaluate the hypothesis that the heavy predation imposed by B. longimanus would have been selected for Daphnia clones that are more negatively phototactic in the presence of B. longimanus cue, because B. longimanus is a visual predator. The behavior of the clones derived from pre-B. longimanus era resting eggs was not significantly different from the behavior of the clones from the post-B. longimanus era and exposure to predator cue did not affect the phototactic response of the clones. There was a significant difference in the phototactic behavior of the three Daphnia species tested (Daphnia ambigua, Daphnia mendotae, and Daphnia pulicaria). These results suggest that predation by B. longimanus is not the main factor that is influencing the phototactic behavior of Daphnia in the lake. Other factors such as fish predation may be playing a more significant role in this system.

Effects of four food dyes on development of three model species, Cucumis sativus, Artemia salina and Danio rerio: Assessment of potential risk for the environment

Food dyes, or color additives, are chemicals added to industrial food products and in domestic cooking to improve the perceived flavor and attractiveness. Of natural and synthetic origin, their safety has been long discussed, and concern for human safety is now clearly manifested by warnings added on products labels. Limited attention, however, has been dedicated to the effects of these compounds on aquatic flora and fauna. For this reason, the toxicity of four different commercially available food dyes (cochineal red E120, Ponceau red E124, tartrazine yellow E102 and blue Patent E131) was assessed on three different model organisms, namely Cucumis sativus, Artemia salina and Danio rerio that occupy diverse positions in the trophic pyramid. The evidence collected indicates that food dyes may target several organs and functions, depending on the species. C. sativus rate of germination was increased by E102, while root/shoot ratio was ∼20% reduced by E102, E120 and E124, seed total chlorophylls and carotenoids were 15–20% increased by E120 and 131, and total antioxidant activity was ∼25% reduced by all dyes. Mortality and low mobility of A. salina nauplii were increased by up to 50% in presence of E124, E102 and E131, while the nauplii phototactic response was significantly altered by E102, E120 and E124. Two to four-fold increases in the hatching percentages at 48 h were induced by E124, E102 and E131 on D. rerio, associated with the occurrence of 20% of embryos showing developmental defects. These results demonstrated that the food dyes examined are far from being safe for the aquatic organisms as well as land organisms exposed during watering with contaminated water. The overall information obtained gives a realistic snapshot of the potential pollution risk exerted by food dyes and of the different organism' ability to overcome the stress induced by contamination.

The Blue-Green Sensory Rhodopsin SRM from Haloarcula marismortui Attenuates Both Phototactic Responses Mediated by Sensory Rhodopsin I and II in Halobacterium salinarum

Haloarchaea utilize various microbial rhodopsins to harvest light energy or to mediate phototaxis in search of optimal environmental niches. To date, only the red light-sensing sensory rhodopsin I (SRI) and the blue light-sensing sensory rhodopsin II (SRII) have been shown to mediate positive and negative phototaxis, respectively. In this work, we demonstrated that a blue-green light-sensing (504 nm) sensory rhodopsin from Haloarcula marismortui, SRM, attenuated both positive and negative phototaxis through its sensing region. The H. marismortui genome encodes three sensory rhodopsins: SRI, SRII and SRM. Using spectroscopic assays, we first demonstrated the interaction between SRM and its cognate transducer, HtrM. We then transformed an SRM-HtrM fusion protein into Halobacterium salinarum, which contains only SRI and SRII, and observed that SRM-HtrM fusion protein decreased both positive and negative phototaxis of H. salinarum. Together, our results suggested a novel phototaxis signalling system in H. marismortui comprised of three sensory rhodopsins in which the phototactic response of SRI and SRII were attenuated by SRM.

Influence of green light illumination on several enzymes involved in energy metabolism in the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae)

Abstract The oriental armyworm, Mythimna separata Walker, is an important insect pest in eastern Asia. Green (520 nm) light with an exposure time of >30 min can result in a stronger phototactic response than the wavelengths of other lights in M. separata moths. The present study was mainly focused to estimate the activities of several enzymes (such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glycerol-3-phosphate dehydrogenase (GPDH), lactate dehydrogenase (LDH) and 3-hydroxyacyl-CoA dehydrogenase (HOAD)) involved in the energy metabolism and a ratio of HOAD to GAPDH activity in M. separata moths exposed to the green light for different exposure times. Our results showed that when M. separata moths were exposed to the green light for 30 min, the activities of the two enzymes (GPDH and HOAD) and as well as the ratio of HOAD to GAPDH activity were significantly elevated. The activity of LDH was significantly increased in the moths exposed for 60 min. Furthermore, significant differences in enzyme activities between the male and female moths were recorded in 45 min exposure time group of the GAPDH, all light treatment groups of the LDH, and 45 min group of the ratio of HOAD to GAPDH activity, respectively. We suggest that when M. separata adult moths are exposed to the green light, these enzymes can be activated to produce energy for starting the phototactic behavior to the green light. Our findings may provide a theoretical basis for elucidating a reason of the phototactic behavior of nocturnal moths.

Shedding light on the effects of hydraulic fracturing flowback and produced water on phototactic behavior in Daphnia magna

The effluent produced during hydraulic fracturing (i.e. flowback and produced water; FPW), is a complex hyper-saline solution that is known to negatively impact the survival and the fitness of the water flea Daphnia magna, but to date effects on behavior are unstudied. In the current study, the effects of FPW on phototactic behavior of D. magna were examined. Exposure of naïve animals to FPW resulted in a dose-dependent increase in the speed of appearance of daphnids in the illuminated zone of the test apparatus (i.e. a faster positive phototaxis response). A similar dose-dependent response was observed in a test solution where the salt content of FPW was recreated in the absence of other components, suggesting that the effect was largely driven by salinity. The effect of FPW was significant when the raw FPW sample was diluted to 20% of its initial strength, while the effect of salt-matched solution was significant at a 10% dilution. A distinct effect was observed following FPW pre-exposure. After a 24 h pre-exposure to 1.5% FPW, Daphnia displayed a significantly inhibited positive phototaxis response when examined in control water, relative to control animals that were not pre-exposed to FPW. This effect was not observed in salinity pre-exposed animals, however these daphnids displayed a significantly reduced phototactic response when tested in saline waters, indicating a loss of the positive phototaxis seen in naïve organisms. These data indicate that FPW can induce perturbations in the behavior of aquatic invertebrates, an effect that may influence processes such as feeding and predation rates.

The role of phototaxis in the initial swim bladder inflation of larval yellow perch (Perca flavescens)

The yellow perch (Perca flavescens) is a high priority species for aquaculture, with strong demand in the USA. Unfortunately, numerous developmental problems during larval rearing prevent its commercial viability. Failed swim bladder inflation (SBI) has been reported in the culture of numerous physoclistous fish species and is a prominent problem in the culture of yellow perch. Initial SBI occurs within a short period of ontogeny, and missing this opportunity results in permanent malformation of the organ. Lighting cues appear to play a role in triggering the response of rising to the surface to swallow the air required for initial SBI in several fishes including yellow perch. Larval yellow perch are photopositive, and this phototactic response seems to co-occur with the window of opportunity for SBI. Thus, we examined the role of phototactic behavior and low-intensity, moonlight-like illumination in initial SBI in yellow perch. Lights were positioned below and above the water surface in larval culture tanks to see if the presence of nighttime illumination impacted SBI success, and if the phototactic behavior of yellow perch helps them find the water surface when inflating at night. The results of this study reveal that even very low-intensity nighttime light reduces the proportion of yellow perch larvae to initially inflate regardless of the direction of the light source. Also, on treatment with no nighttime illumination, higher than expected SBI was observed, and the culture conditions that produced this success should be examined when focusing on improving SBI in the future. This research suggests that the photopositive response of yellow perch is not involved in SBI success, and lighting could play a role in hindering the process.

Optical toxicity of triphenyl phosphate in zebrafish larvae

Triphenyl phosphate (TPhP) has been shown to cause developmental neurotoxicty. Considering the visual system is a sensitive target, in the present study, we investigated the potential toxicity of TPhP on the visual development and function in zebrafish larvae. Embryos were exposed to 0, 0.1, 1, 10, and 30 μg/L TPhP from 2 to 144 h post-fertilization (hpf). The transcription of photoreceptor opsin genes, and histopathological changes in the retina and visual behavior (optokinetic and phototactic responses) were evaluated. TPhP significantly downregulated the transcription of opsin genes (zfrho, opn1sw1, opn1sw2, opn1mw1, opn1mw2, opn1mw3, opn1mw4, opn1lw1 and opn1lw2) in all exposure groups. Histopathological analysis revealed that the areas of the outer nuclear layer (ONL), inner nuclear layer (INL), and inner plexiform layer (IPL) of the retina were significantly reduced in the 10 and 30 μg/L TPhP groups. The number of ganglion cells was reduced significantly in the 30 μg/L group. The optokinetic response (OKR) and phototactic response showed dose-dependent decreases caused by impaired visual function, which was confirmed by unchanged locomotor activity. The results indicated that exposure to environmentally relevant concentrations of TPhP could inhibit the transcription of genes related to visual function and impair retinal development, thus leading to visual impairment in zebrafish larvae.

Volatiles mediate host-selection in the corn hoppers Dalbulus maidis (Hemiptera: Cicadellidae) and Peregrinus maidis (Hemiptera: Delphacidae).

Volatile organic compounds (VOCs) released by plants are generally involved in host recognition and host selection for many phytophagous insects. However, for leafhoppers and planthoppers, host recognition is mainly thought to involve a phototactic response, but it is not clear if a host plant could be selected based on the volatile cues it emits. In this study we evaluated olfactory responses in dual choice tests of two Hemiptera species, Dalbulus maidis (De Long) (Cicadellidae) and Peregrinus maidis (Ashmead) (Delphacidae), vectors of maize-stunting diseases, to three maize (Zea mays L.) germplasms, a temperate and a tropical hybrid and a landrace. VOCs emitted by the germplasms were collected and identified using gas chromatography-mass spectrometry. The temperate hybrid released significantly more VOCs than the tropical hybrid and the landrace, and its volatile profile was dominated by (±)-linalool. D. maidis preferred odours emitted from the temperate hybrid, whereas P. maidis preferred odours from the tropical hybrid and the landrace over the temperate one. In order to test if linalool plays a role in the behavioural responses, we assayed this compound in combination with the tropical hybrid, to provide other contextual olfactory cues. D. maidis was attracted to the tropical hybrid plus a 0.0001% linalool solution, indicating that this compound could be part of a blend of attractants. Whereas addition of linalool resulted in a slight, though not significant, reduction in host VOC attractiveness for P. maidis. Both hopper species responded to olfactory cues in the absence of supplementary visual cues.

Anesthetic protocol for microinjection-related handling of Siberian sturgeon (Acipenser baerii; Acipenseriformes) prolarvae.

An anesthetic protocol was optimized for microinjection-related handling of Siberian sturgeon (Acipenser baerii; Acipenseriformes) prolarvae, an extant primitive fish species commonly grown in aquaculture. Comparative examinations of three selected anesthetics (clove oil, lidocaine, and MS-222) with a dosage regime of 50, 100, 200, and 400 mg/L indicated that MS-222 was the most efficient agent for Siberian sturgeon prolarvae, as evidenced by the fast induction of anesthesia with quick and uniform recovery. Meanwhile, clove oil should be avoided, due to prolonged recovery times varying widely between individuals. None of the tested anesthetics significantly affected prolarval viability at any of the dosage regimes tested in this study. Based on an analysis of the duration of an unconscious state in air, we recommend a dose of 200 mg/L MS-222 for microinjection. Recovery time after use of this dose was influenced by the prolarval age and the development of gills, in which prolarvae older than 3 days after hatching required longer recovery times than did younger prolarvae. Post-recovery behavioral assessment showed no apparent difference between MS-222-anesthetized and non-anesthetized prolarvae in their swimming behavior and phototactic responses. Applicability of currently developed anesthetic protocol using MS-222 in larval microinjection was demonstrated with the injection of a visible dye to the anesthetized prolarvae, followed by the analysis of post-recovery viability. Taken together, the present anesthetic protocol based on 200 mg/L of MS-222 could provide researchers with practical usefulness with good safety margins for the micromanipulation and other related handlings of Siberian sturgeon prolarvae.

Effect of Different Light Spectrum in Helicoverpa armigera Larvae during HearNPV Induced Tree-Top Disease.

Lepidopteran larvae upon infection by baculovirus show positive photo-tactic movement during tree-top disease. In light of many insects exploiting specific spectral information for the different behavioral decision, each spectral wavelength of light is an individual parsimonious candidate for such behavior stimulation. Here, we investigated the responses of third instar Helicoverpa armigera larvae infected by Helicoverpa armigera nucleopolyhedrovirus (HearNPV) to white (broad-spectrum), blue (450–490 nm), UVA (320–400 nm), and UVB (290–320 nm) lights for the tree-top disease. Our findings suggest that tree-top phenomenon is induced only when the light is applied from above. Blue, white and UVA lights from above induced tree-top disease, causing infected larvae to die in an elevated position compared to those larvae living in the complete dark. In contrast, UVB from above did not induce tree-top disease. Blue light exerted the maximum photo-tactic response, significantly (p < 0.01) higher than white light. The magnitude of the response decreased with decreasing wavelength to UVA, and no response at UVB. Our results suggested that the spectral wavelength of the light has a significant effect on the induction of the tree-top disease in H. armigera third instar larvae infected with HearNPV.

Zebrafish larvae show negative phototaxis to near-infrared light.

Zebrafish larvae (Danio rerio) are among the most used model species to test biological effects of different substances in biomedical research, neuroscience and ecotoxicology. Most tests are based on changes in swimming activity of zebrafish larvae by using commercially available high-throughput screening systems. These systems record and analyse behaviour patterns using visible (VIS) and near-infrared (NIR) light sources, to simulate day (VIS) and night (NIR) phases, which allow continuous recording of the behaviour using a NIR sensitive camera. So far, however, the sensitivity of zebrafish larvae to NIR has never been tested experimentally, although being a critical piece of information for interpreting their behaviour under experimental conditions. Here, we investigated the swimming activity of 96 hpf (hours post fertilization) and 120 hpf zebrafish larvae under light sources of NIR at 860 nm and at 960 nm wavelength and under VIS light. A thermal source was simultaneously presented opposite to one of the light sources as control. We found that zebrafish larvae of both larval stages showed a clear negative phototactic response towards 860 nm NIR light and to VIS light, but not to 960 nm NIR light. Our results demonstrated that zebrafish larvae are able to perceive NIR at 860 nm, which is almost identical to the most commonly used light source in commercial screening systems (NIR at 850 nm) to create a dark environment. These tests, however, are not performed in the dark from the zebrafish´s point of view. We recommend testing sensitivity of the used test organism before assuming no interaction with the applied light source of commonly used biosensor test systems. Previous studies on biological effects of substances to zebrafish larvae should be interpreted with caution.

Two-Dimensional Microfluidic System for the Simultaneous Quantitative Analysis of Phototactic/Chemotactic Responses of Microalgae.

Microalgae have been spotlighted as a renewable energy source to produce biofuels from CO2 by photosynthesis. However, their innate inefficiency of CO2 conversion using light energy has been a challenge to the commercialization of algae-based biofuel production. Photosynthetic organisms have evolved behavioral responses, including phototaxis and chemotaxis, to find optimal conditions for capturing light energy and inorganic carbon (Ci) sources for photosynthesis. In this context, investigation of phototaxis and chemotaxis to HCO3-, the predominant form of Ci in neutral aqueous solutions, is necessary to understand the physiological role of tactic responses in photosynthesis. In this study, a two-dimensional microfluidic system enabled efficient analysis of phototactic and chemotactic responses by investigation of cell distribution in the outlet chambers. From statistical analysis (skewness and kurtosis) of tactic responses of different algal strains to external stimuli, the preferred concentrations of HCO3- for Chlamydomonas reinhardtii CC125 (wild type), PTS42 (random insertional mutant of C. reinhardtii, high photosynthetic activity), and CC2702 ( cia5 mutant of C. reinhardtii, unable to acclimate to low CO2 concentration) were determined to be 27.22, 43.23, and 36.95 mM, respectively. From the analysis of tactic responses of wild type and 14 mutant strains, it was found that the photosystem II (PSII) operating efficiency and CO2 fixation rate were strongly correlated with the phototactic ( R2 = 0.931) and chemotactic response ( R2 = 0.857), respectively. Finally, this system can be applied to high-throughput screening strategies for the rapid isolation of high photosynthetically productive microalgal strains based on their tactic responses.