Shoaling behaviour in a surface-dwelling and a cave-dwelling population of a barb <Emphasis Type="Italic">Garra barreimiae</Emphasis> (Cyprinidae,Teleostei)

We studied shoaling behaviour in a species of fish (Garra barreimiae) from Oman. We compared two populations (a surface-dwelling and a cave-dwelling population) with different theoretical costs and benefits of shoaling. We measured the tendency to associate with a shoal of conspecifics. The stimulus shoal was confined to (1) clear Plexiglas cylinders in light, (2) wire-mesh cylinders in light, or (3) wire-mesh cylinders in darkness. The surface form exhibited a strong preference for the stimulus shoal during the experiments in light, but also in darkness, when only non-visual cues from the shoal could be perceived. The cave form did not show a preference when solely visual cues were available (Plexiglas cylinder). When non-visual cues from the shoal could be perceived (wire-mesh), the cave form did show a preference to associate with the shoal, but the shoaling tendency was considerably weaker than in the surface form. The shoaling tendency has probably been genetically reduced in the cave form.     

Light-adaptation in the photophobic response by Stentor coeruleus

Effects of preillumination on photophobic response (light-adaptation) and recovery of the photophobic sensitivity in the dark (dark-adaptation) in Stentor coeruleus were examined. When the cells were preilluminated with white light of 7.80 W/m² for 2 min, the fluence-rate response curve of photophobic response was shifted toward higher light intensities by half an order of magnitude compared to the one without preillumination. Preillumination with a higher light intensity resulted in a further shift of the fluencerate response curve. An action spectrum for light-adaptation showed a primary peak at 610 nm and secondary peaks at 540 and 480 nm which are almost identical to the peaks observed in the photophobic action spectrum.The light-adapted cells showed a recovery of their photophobic sensing ability following dark treatment. Dark-adaptation resulted in total recovery of photophobic sensing ability in 8 minutes for the most cases examined.     

Offspring number in a livebearing fish (<Emphasis Type="Italic">Poecilia mexicana</Emphasis>, Poeciliidae): reduced fecundity and reduced plasticity in a population of cave mollies

Life history traits within species often vary among different habitats. We measured female fecundity in mollies (Poecilia mexicana) from a H₂S-rich cave and from a neighbouring surface habitat, as well as in laboratory-reared individuals of both populations raised in either light or continuous darkness. Compared to conspecifics from surface habitats, cave-dwelling P. mexicana had reduced fecundity (adjusted for size) in the field. In the laboratory, the fecundity of surface mollies was higher in light than in darkness, whereas fecundity in the cave mollies was almost unaffected by the ambient light conditions. Our results suggest a heritable component to the reduction in fecundity in female cave mollies. Moreover, the reduced plasticity in fecundity of cave mollies in response to light conditions might be an example of genetic assimilation or channelling of a life history trait in a population invading a new environment.     

Photomovement in Dunaliella salina: Fluence rate-response curves and action spectra

We determined the action spectra of the photophobic responses as well as the phototactic response in Dunaliella salina (Volvocales) using both single cells and populations. The action spectra of the photophobic responses have maxima at 510 nm, the spectrum for phototaxis has a maximum at 450–460 nm. These action spectra are not compatible with the hypothesis that flavoproteins are the photoreceptor pigments, and we suggest that carotenoproteins or rhodopsins act as the photoreceptor pigments. We also conclude that the phototactic response in Dunaliella is an elementary response, quite independent of the step-up and step-down photophobic responses. We also determined the action spectra of the photoaccumulation response in populations of cells adapted to two different salt conditions. Both action spectra have a peak a 490 nm. The photoaccumulation response may be a complex response composed of the phototactic and photophobic responses. Blue or blue-green light does not elicit a photokinetic response in Dunaliella.Diagrams of the optical set-ups used for measuring the responses at the single-cell level and of the plans for building the phototaxometer described in this paper are available to the interested readerWe thank Mr. M. Kubota for a tremendous amount of technical assistance and Mr. R. Nagy for building the phototaxometer. We thank T. Kondo, Professor H. Imaseki and the members of the Laboratory of Biological Regulation, NIBB, for their help and support in various aspects of this research. This research was supported, in part, from grants from the Okazaki Large Spectrograph (Project Nos. 86-535, 87-518, 88-523), the Japanese Society for the Promotion of Science, and the College of Agriculture and Life Sciences at Cornell University to R. W.     

光谱和光强度对棕榈蓟马雌成虫行为反应的影响

室内研究了光谱、光强度对棕榈蓟马雌成虫的趋、避光行为的影响。结果显示:在340—605 nm波谱内棕榈蓟马雌成虫对14个单色光刺激的趋光行为反应为多峰型。其中蓝光483 nm处峰最高,趋光反应率达34.96%,其次为绿光498—524 nm、562—582 nm、紫外光340 nm处;其避光行为反应共有3个峰,其中紫外光380 nm处最高,避光率18.08%,另外2个峰分别在橙光605 nm、紫光420 nm处。在趋光率较高的单色光(340、483、524、582 nm)和避光率较高的单色光(380、605 nm)以及白光刺激下,棕榈蓟马雌成虫的趋光率随光强增强的增强而提高,而避光率则随着光强的增强而降低;光强最弱时仍均有一定趋光率,最强时均未出现高端平台。因此:棕榈蓟马雌成虫对不同单色光具有明显的选择性,光谱和光强度对其趋光行为和避光行为有较大影响,光强度的影响作用与波长因素有关。     

Characterization of the Eyespot Regions of "Blind"Chlamydomonas Mutants after Restoration of Photophobic Responses

Chlamydomonas reinhardtii exhibits photophobic and positive and negative phototactic responses that can be defined for cell populations using computerized cell tracking and motion analysis. Mutants CC-2359 and FN68 are pigment deficient mutants that are blocked in carotenoid synthesis and lack these photo responses. In particular, neither mutant exhibits flash-induced photophobic responses to visible light stimuli to which wild-type gametic cells exhibit a strong response, with several behavioral stages. Upon addition of all-trans retinal to these mutants, the photophobic responses are restored with minor quantitative differences from wild-type populations. Using both light and electron microscopy, we have compared the ultrastructural characteristics of wild-type C. reinhardtii to those of both mutants. As previously described, wild-type cells contain an eyespot consisting of 2–4 layers of pigmented granules encased within thylakoid membranes, located between the distal extremities of the flagellar root. This structure is also visible as an orange-red spot in light microscopy. The photoreceptor is thought to be concentrated in the plasma membrane above the eyespot. The mutant, CC-2359, lacks this eyespot as seen by both light and electron microscopy, even when the photophobic response has been restored. FN68-like mutants studied earlier by Morel-Laurens and Feinlieb and others contain an eyespot which can be seen only by electron microscopy. In FN-68, the eyespot generally has the same dimensions as in wt cells, differing mainly in pigment granule appearance. Consistent with these findings, several laboratories have shown that the full range of phototactic responses can be reconstituted in FN68 and CC-2359, but that negative phototaxis requires a significantly stronger light stimulus in the latter strain. We confirm the suggestion that the eyespot is not necessary for the photophobic response, and is not critical for the appropriate assembly and function of the photophobic response receptor in the membrane. Furthermore, the locus of reconstitution of the functional receptor is not the eyespot. Because of the definitive demonstration of the absence of the eyespot in CC-2359, however, the eyespot may play a role in negative phototaxis.     

Anion sensitivity of motility and step-down photophobic responses of Euglena gracilis

The motility and step-down photophobic responses of Euglena are influenced by inorganic and organic anions. Persistent motility (with Ca²⁺, Mg²⁺ and K⁺ present) is supported with chloride or sulfate but not with acetate, nitrate or propionate as the only added anions. Cells in media containing acetate displayed a cell aggregation (clumping) behavior that was both red light sensitive and, under some conditions, was accompanied by suppression of the step-down photophobic response. Addition of sodium salts (Cl^-, SO ₄ ^2- , acetate or propionate) to cells in Cl^- or SO ₄ ^2- based media had differential effects on the duration of the step-down photophobic responses induced by blue light removal: anions alter the response. In addition, cells in all Cl^- containing media showed constant photophobic response duration following repeated stimulation. Cells in some SO₄ ^2- containing media, however, showed response summation to repeated stimulation. This latter effect was reversible and was overcome by the addition of chloride anions.     

Parallel evolution leads to reduced shoaling behavior in two cave dwelling populations of Atlantic mollies (<Emphasis Type="Italic">Poecilia mexicana</Emphasis>, Poeciliidae,Teleostei)

Shoaling behavior protects fishes from avian and piscine predation, but at the same time costs of group living arise due to several mechanisms including increased food competition. Most cave fishes live in an environment in which avian and piscine predators are lacking, and cave environments are often characterized by low food availability, leading to increased food competition. Altogether, this should favor the reduction of shoaling in cave fishes. We compared shoaling behavior (i.e. the tendency to associate with a stimulus shoal) among surface dwelling populations of the Atlantic molly, Poecilia mexicana, and two cave forms of that species. The first cave population of P. mexicana originated from the Cueva del Azufre and was previously recognized as the only cave form of a poeciliid fish. The second cave population examined came from a cave that was discovered only recently (Cueva Luna Azufre). In both cave forms shoaling behavior was reduced compared with surface dwelling mollies.     

Standing genetic variation as a potential mechanism of novel cave phenotype evolution in the freshwater isopod,Asellus aquaticus

Novel phenotypes can come about through a variety of mechanisms including standing genetic variation from a founding population. Cave animals are an excellent system in which to study the evolution of novel phenotypes such as loss of pigmentation and eyes. Asellus aquaticus is a freshwater isopod crustacean found in Europe and has both a surface and a cave ecomorph which vary in multiple phenotypic traits. An orange eye phenotype was previously revealed by F₂ crosses and backcrosses to the cave parent within two examined Slovenian cave populations. Complete loss of pigmentation, both in eye and body, is epistatic to the orange eye phenotype and therefore the orange eye phenotype is hidden within the cave populations. Our goal was to investigate the origin of the orange eye alleles within the Slovenian cave populations by examining A. aquaticus individuals from Slovenian and Romanian surface populations and Asellus aquaticus infernus individuals from a Romanian cave population. We found orange eye individuals present in lab raised surface populations of A. aquaticus from both Slovenia and Romania. Using a mapping approach with crosses between individuals of two surface populations, we found that the region known to be responsible for the orange eye phenotype within the two previously examined Slovenian cave populations was also responsible within both the Slovenian and the Romanian surface populations. Complementation crosses between orange eye Slovenian and orange eye Romanian surface individuals suggest that the same gene is responsible for the orange eye phenotype in both surface populations. Additionally, we observed a low frequency phenotype of eye loss in crosses generated between the two surface populations and also in the Romanian surface population. Finally, in a cave population from Romania, A. aquaticus infernus, we found that the same region is also responsible for the orange eye phenotype as the Slovenian cave populations and the Slovenian and Romanian surface populations. Therefore, we present evidence that variation present in the cave populations could originate from standing variation present in the surface populations and/or transgressive hybridization of different surface phylogenetic lineages rather than de novo mutations.     

The role of photoreception in the swarming behavior of the copepod Dioithona oculata

The copepod Dioithona oculata forms dense swarms near mangrove prop roots that are centered around shafts of light penetrating the mangrove canopy. Swarms can be created in the laboratory within light shafts created with a fiber optic light pipe. Laboratory observations of swarming behavior were recorded using video cameras, and the swimming behavior of the copepods and density of the swarms were quantified using video‐computer motion and image analysis techniques. Swarm formation results from a combination of phototactic and klino‐kinetic behavior. Dark adapted copepods initially exhibit a photophobic response to a light shaft, but become positively phototactic within 3–5 min after exposure to the light. Copepod aggregation rates under the light fit a saturation model, suggesting that copepods are attracted independently to the swarm marker. Copepods reverse their swimming direction when they encounter light intensity gradients near the edge of a light shaft, which aids in maintaining the swarm. Swarm formation can occur in the laboratory at light intensities as slow as 0.1 μM photons m^‐2 s^‐1, which is similar to light intensities at dawn when they are first observed to form in nature. Swarm formation appears to have an endogenous rhythm, as copepods will not form swarms at night under a light shaft.     

Is foraging innovation lost following colonization of a less variable environment? A case study in surface‐ vs. cave‐dwelling Asellus aquaticus

Behavioral innovation is a key process for successful colonization of new habitat types. However, it is costly due to the necessary cognitive and neural demands and typically connected to ecological generalism. Therefore, loss of behavioral innovativeness is predicted following colonization of new, simple, and invariable environments. We tested this prediction by studying foraging innovativeness in the freshwater isopod Asellus aquaticus. We sampled its populations along the route of colonizing a thermokarstic water‐filled cave (simple, stable habitat with only bacterial mats as food) from surface habitats (variable environment, wide variety of food). The studied cave population separated from the surface populations at least 60,000 years ago. Animals were tested both with familiar and novel food types (cave food: bacterial mats; surface food: decaying leaves). Irrespective of food type, cave individuals were more likely to feed than surface individuals. Further, animals from all populations fed longer on leaves than on bacteria, even though leaves were novel for the cave animals. Our results support that cave A. aquaticus did not lose the ability to use the ancestral (surface) food type after adapting to a simple, stable, and highly specialized habitat.     

Gene flow and population structure in the Mexican blind cavefish complex (<Emphasis Type="Italic">Astyanax mexicanus</Emphasis>)

Background  

Cave animals converge evolutionarily on a suite of troglomorphic traits, the best known of which are eyelessness and depigmentation. We studied 11 cave and 10 surface populations of Astyanax mexicanus in order to better understand the evolutionary origins of the cave forms, the basic genetic structuring of both cave and surface populations, and the degree to which present day migration among them affects their genetic divergence.     

Reduction in the metabolic levels due to phenotypic plasticity in the Pyrenean newt,Calotriton asper,during cave colonization

According to theories on cave adaptation, cave organisms are expected to develop a lower metabolic rate compared to surface organisms as an adaptation to food scarcity in the subterranean environments. To test this hypothesis, we compared the oxygen consumption rates of the surface and subterranean populations of a surface‐dwelling species, the newt Calotriton asper, occasionally found in caves. In this study, we designed a new experimental setup in which animals with free movement were monitored for several days in a respirometer. First, we measured the metabolic rates of individuals from the surface and subterranean populations, both maintained for eight years in captivity in a natural cave. We then tested individuals from these populations immediately after they were caught and one year later while being maintained in the cave. We found that the surface individuals that acclimated to the cave significantly reduced their oxygen consumption, whereas individuals from the subterranean population maintained in the cave under a light/dark cycle did not significantly modify their metabolic rates. Second, we compared these metabolic rates to those of an obligate subterranean salamander (Proteus anguinus), a surface aquatic Urodel (Ambystoma mexicanum), and a fish species (Gobio occitaniae) as references for surface organisms from different phyla. As predicted, we found differences between the subterranean and surface species, and the metabolic rates of surface and subterranean C. asper populations were between those of the obligate subterranean and surface species. These results suggest that the plasticity of the metabolism observed in surface C. asper was neither directly due to food availability in our experiments nor the light/dark conditions, but due to static temperatures. Moreover, we suggest that this adjustment of the metabolic level at a temperature close to the thermal optimum may further allow individual species to cope with the food limitations of the subterranean environment.     

Morphological Alterations in Euglena gracilis Induced by Treatment with CTAB (Cetyltrimethylammonium Bromide) and Triton X-100: Correlations with Effects on Photophobic Behavioral Responses*

SYNOPSIS. Treatment of Euglena gracilis with the cationic detergent CTAB at concentrations of 0.05 mM or higher selectively inhibited the ability of the cells to respond with flagellar reorientation to a sudden decrease of light intensity (step-down photophobic response). The ability to respond similarly to an increase in light intensity (step-up photophobic response) was unaffected even at detergent concentrations at which the step-down response was completely inhibited. Electron microscopy of cells treated with 1.0 mM CTAB revealed selective destruction of the membrane of the reservoir and flagellum. No selective effects upon the step-down or step-up photophobic responses were found upon treatment of the cells with Triton X-100.     

Irradiance Regulation of Photosynthesis and Respiration in Modern Marine Microbialites Built by Benthic Cyanobacteria in a Tropical Lagoon (New Caledonia)

Microbialites are organosedimentary deposits that have built up as a result of the growth and binding of detrital sediment by a benthic microbial community. This study focuses on microbialites built by monospecific populations of cyanobacteria in the south-west lagoon of New Caledonia, where they have been observed down to 20–25 m depth. The aim was to study their photosynthetic and respiratory responses to various light intensities. The Phormidium sp. TK1 microbialite was collected at 19 m depth and the P. crosbyanum (Tilden) microbialite was collected at 0.5 and 13 m depth. Phormidium sp. TK1 showed all the characteristic features of a low-light adapted species. The initial slope of the Photosynthesis versus Irradiance curve for this microbialite was close to the maximum quantum yield indicating an efficient light absorption and utilization at low light. The photosynthesis maximum was located 0.2–0.4 mm below the surface and did not shift with changing light intensity. Respiration rates were low and not enhanced by light; photoinhibition was observed at higher light intensities. In Phormidium crosbyanum (Tilden) microbialites, the photosynthesis maximum shifted downward to lower depths with increasing light, probably as a result of phototactic migration of cyanobacterial filaments, and light-enhanced respiration was observed at light intensities above light saturation. The photosynthetic para- meters measured in P. crosbyanum indicate that P. crosbyanum is capable of photo-acclimation at high light intensities. The gross productivity of the different microbialites was comparable to values measured in cyanobacterial stromatolites observed in other shallow environments. However, the microbialites studied here were characterized by a lower respiration / production ratio which indicates a higher growth efficiency.     

Random Amplified Polymorphic DNA Diversity among Surface and Subterranean Populations of Asellus aquaticus (Crustacea: Isopoda)

The ecological and evolutionary processes leading to isolation and adaptation of cave animals compared to their surface ancestors are not yet unequivocally understood. In this study the genetic relations of four cave and three surface population of the freshwater crustacean Asellus aquaticus in the Karst region of SW Slovenia and NE Italy were assessed using RAPDs as genetic markers. The results suggest that specialized populations from two caves invaded their subterranean habitat independently, and that their morphological similarity is a result of convergent evolution. Another, less specialized cave population seems to originate from a later colonization of a cave system already inhabited by a more specialized population, but the two populations do not interbreed. This series of temporally and spatially independent invasions has generated a diversity hotspot of non-interbreeding populations of a ubiquitous freshwatercrustacean, which is uniform over most of its range. Genetic variability estimated by the percentage of polymorphic RAPD fragments was similar (86–91%) in most cave and surface populations. Substantially lower values (as low as 49%) were found in two cave populations affected by heavy pollution. Two a priori groupings of populations, traditional subspecies and hydrologically connected groups, were rejected as not significant by means of nested analysis of molecular variance (AMOVA). On the other hand, groupings revealed by UPGMA clustering displayed a significant component of among-group variance. An analysis of gene flow between populations using estimated migration rates confirmed these findings.     

Origins and Relationship of Cave Populations of the Blind Mexican Tetra, Astyanax Fasciatus, in the Sierra De El Abra

The blind morph of Astyanax fasciatus (Pisces: Characidae) has been more thoroughly studied than any other cave inhabiting organism. Most studies of A. fasciatus have used individuals from different caves of the Sierra de El Abra, Mexico, and have assumed that each population independently evolved to live in the cave environment. We analyzed the relationships among several cave populations that delineate the Sierra de El Abra using RAPD markers. The results indicate that all cave populations are more closely related to one another than they are to the surface populations. This suggests that present day cave populations derived from a common ancestral stock, most likely due to a single colonization event, or alternatively, that strong gene flow among cave populations has occurred, raising precaution against considering each cave population as independent.     

Photophobic responses of calanoid copepods: possible adaptive value

A genual pattern of photophobic responses has been observedwhich differs for calanoid copepods from freshwater, estuarineand oceanic environments. Using a video-computer system formotion analysis, the photophobic responses of light and darkadapted calanoid copepods were compared. Dark-adapted copepodswere exposed to 600 ms flashes of dim blue light at 5 s intervalswhich simulated the flashes of biolumines-cent marine zooplankton.Light-adapted copepods were exposed to 600 ms intervals of darknessat 5 s intervals to simulate the shadows of organisms passingoverhead. Four species of coastal marine copepods (Acartia hudsonica,Centropages hamatus, Pseudocalanus minutus and Temora longicornis)all showed photophobic responses to both flashes and shadows.These responses may have adaptive value to the copepods sincethey live in an environment with predators that are bioluminescentat night and cast shadows on their prey during the day (e.g.ctenophores and cnidarian medusae). Two species of oceanic copepods(Euchaeta marina, Pleuromamma abdominalis) showed strong photophobicresponses to flashes but no response to shadows. This may correspondto the abundance of bioluminescent predators on copepods inthe oceanic environment (fish, ctenophores, siphonophores, etc.)and their lack of exposure to the shadows of predators, sinceboth these species are rarely found in the euphoric zone duringthe day. Two species of freshwater copepods (Diaptomus sanguineus,Epishwa massachusettsensis) showed no similar photophobic responseto flashes of light. This lack of startle response may relateto the lack of bioluminescence in the freshwater environment.Freshwater copepods showed a weak photophobic response to shadows.The adaptive value of this behavior is unclear, however, sincethe responses seem to be too weak to function for escape, andthe dominant predators large enough to cast shadows (fish) tendto approach their prey laterally. ¹Present address: Marine Science Institute, University of Texasat Austin, Port Aransas, TX 78373–1267, USA     

Step-up photophobic response in the ciliate,Stentor coeruleus

The avoidance by Stentor coeruleus of a light trap is caused by a step-up photophobic response. The phobic response invariably consists of a delay of about 200 ms, a stop response, a turn to one side, and resumption of swimming in the new direction. After this the cells enter a refractory period of 1–3 s following a phobic response, during which they will not give a second response. Phobic responses can be elicited by spatial and temporal increases in light intensity. The action spectrum for the step-up photophobic response resembles the absorption spectrum of stentorin, the proposed photoreceptor pigment, and of its chromophore, hypericin.The phobic response is specifically inhibited by the protonophorous uncouplers TPMP⁺ and FCCP but not by the ionophores gramicidin and A23187. Since the uncouplers block light-induced membrane potential changes at the same concentrations, it has been proposed that the primary photoreception causes a light-induced potential change, which in turn, induces a motor response.Abbreviations TPMP⁺ triphenyl methyl phosphonium bromide - FCCP carbonylcyanide p-trifluoromethoxy-phenylhydrazone     

Photophobic responses of the cyanobacterium Anabaena variabilis

The photophobic responses in the Cyanobacterium Anabaena variabilis which belongs to the Nostocaceae have been studied with aid of a population method as well as by single trichome observations. In white light experiments both step-up and step-down photophobic responses were observed. The wavelength dependence was examined at a constant fluence rate. The photophobically active light is absorbed by the photosynthetic pigments, mainly by the phycobiliproteins and chlorohyll a. Above 690 nm only negative reactions were observed, i.e. the trichomes left the light trap. In white light experiments DCMU strongly inhibited the photophobic responses, whereas photokinesis was not affected to the same extent indicating that the reaction is coupled with the non cyclic photosynthetic electron transport. DBMIB impaired the photophobic behaviour only slightly. It seems that the photophobic responses of A. variabilis are controlled by a similar mechanism as in Phormidium uncinatum (Oscillatoriaceae) although the two families and, hence, the two species differ in their movement mechanism as well as in their photoactic behaviour.