Latitudinal trends in size-dependence of bioluminescence in the midshipman fish Porichthys notatus

The bioluminescent fish Porichthys notatus (plainfin midshipman), has a discontinuous distribution along the Pacific coast of North America. The fish is present from Cape Mendocino southward to Baja California, Mexico, absent off the coast of Oregon, USA, and abundant, northward, in Puget Sound, Washington. Interestingly, the population in Puget Sound lacks the substrate (luciferin) necessary for the luminescence reaction and, despite possessing an otherwise fully functional photophore system, is nonluminescent. The California population of P. notatus is uniformly luminescent south of Monterey Bay, but 15% of the speciments tested from San Francisco Bay and the Gulf of the Farallons have been reported to be nonluminescent. Explanations for nonluminescent midshipman in both Puget Sound and the San Francisco Bay area have focussed on a dietary requirement for luciferin. To gain further insight into reasons for nonluminescence in the San Francisco Bay region, the distribution of bioluminescence in P. notatus was studied from Monterey Bay to Cape Mendocino during 1985. A complex pattern of bioluminescence was found, in which nonluminescent individuals reflected neither a local anomaly in the San Francisco Bay region nor a simple gradient of decreasing luminescence towards the northern end of the range of the California population. Instead, a distinct size-dependent component in luminescence capability of the fish was observed. Aspects of the life history of P. notatus and related factors which might influence the bioluminescence characteristics of this population are discussed.Please address all correspondence to Dr. F.I. Tsuji at the Osaka Bioscience Institute     

Disruptive counterillumination and its anti-predatory value in the plainfish midshipman Porichthys notatus

The plainfin midshipman Porichthys notatus Girard occurs in nearshore waters off the western coast of North America. We found that the ventrally directed photophores of juvenile laboratory-grown P. notatus function in counterillumination. Their bioluminescence matched downwelling light to obscure the silhouette as viewed from below. P. notatus did not obligatorily counterilluminate at all times in appropriate light. A hydrodynamic stimulus, in addition to appropriate downwelling light, was required to induce counterillumination. This is perhaps an adaptation to conserve its Vargula-type luciferin, which is obtained from its diet. The angular distribution of light produced by P. notatus approximated that typical of downwelling light in the ocean. There was a direct relationship between downwelling irradiance and the maximum intensity of P. notatus luminescence. Insufficient light was produced to replace the light blocked by the fish under lighting conditions typical of the natural environment. The luminescence was, however, effective in eliminating the silhouette of P. notatus as observed with an image-intensifying camera and by dark-adapted human observers. Consequently, the effectiveness of counterillumination probably originates in part from the disruptive pattern of the luminous photophores. P. notatus from the Puget Sound population, which lacks a luciferin source and is non-luminous, displayed counterillumination behavior when its luminescent system was activated by force-feeding the fish with dried Vargula hilgendorfi. Experiments using adult P. notatus as predators on a mixed population of non-luminous (Puget Sound) and luminous (Santa Barbara Channel) juvenile P. notatus revealed that under conditions of dim overhead illumination non-luminous P. notatus were preyed upon at twice the rate of luminous individuals. This is the first experimental evidence suggesting that counterillumination is effective as an anti-predatory strategy. Received: 25 August 1998 / Accepted: 25 March 1999     

Vision in the lanternfish Stenobrachius leucopsarus (Myctophidae)

The visual system of the midwater fish Stenobrachius leucopsarus (Myctophidae) has been studied by biochemical, anatomical, and electrophysiological methods. Partial bleaching analysis in the presence of hydroxylamine showed that the eye contains a single extractable photopigment, based on retinal and absorbing maximally at 492 nm. The photoreceptor population consists entirely of rods, approximately 5.0×10⁵ rods per mm² and 1.8×10⁷ per retina. Visual sensitivity is enhanced by the lack of pigment granules in the pigment epithelium and the presence of a choroidal tapetum lucidum. Influenced by the high concentration of visual pigment, the spectral sensitivity has a broad plateau between 460 and 540 nm, which suggests that the eye retains high sensitivity to a diversity of bioluminescent stimuli and depth-attenuated solar light.     

Correlation of bioluminescence emissions with ventral photophores in the mesopelagic squidAbralia veranyi (Cephalopoda: Enoploteuthidae)

Specimens of the squidAbralia veranyi were collected off the Bahamas in 1989. The bioluminescence of the ventral photophores was recorded on videotape at 11 to 12°C and 24°C, in conjunction with measurements of the spectral emission at the same temperatures. At 11 °C, the spectrum was unimodal, peaking at 490 nm with a narrow bandwidth. At the higher temperature a shoulder at about 440 nm appeared in the emission spectrum. The short, bright flashes from the subocular photophores had a broader bandwidth and a shorter wavelength emission maximum (475 nm) than that of the ventral photophores. Video recordings at the two temperatures showed no changes in either the identities of the luminescing photophores or their relative intensities. One structurally distinct type of photophore was not illuminated. We conclude that the observed spectral changes were produced within one group of photophores, and that recruitment of the unilluminated photophores would produce an additional spectral component, previously reported from another species ofAbralia (A. trigonura).     

Two populations of the marine fish Porichthys notatus,one lacking in luciferin essential for bioluminescence

Porichthys notatus is a common benthic fish found along the Pacific coast of North America. It possesses more than 700 small dermal photophores on its head and trunk. When P. notatus collected in Monterey Bay, California, is injected subcutaneously with norepinephrine, the photophores emit a long-lasting luminescence that is readily visible to the dark-adapted eye. The light emission is due to the oxidation of luciferin (substrate) by molecular oxygen, catalyzed by luciferase. In contrast, P. notatus collected in Puget Sound, Washington, is nonluminous, even though the photophores do not differ ultrastructurally from those of the California fish. The inability of the Puget Sound fish to luminesce is due to lack of luciferin in its photophores. Luminescence capability, however, may be induced in the Puget Sound fish by the oral or intraperitoneal administration of a small amount of luciferin from the tiny luminescent marine ostracod crustacean, Vargula hilgendorfii, suggesting that the origin of luciferin in P. notatus may be from the diet. In this study, specimens of P. notatus were collected over the entire known range of the species, between November 1981 and September 1987, and the presence of luciferin in the photophores determined. The results indicate that there are two populations of P. notatus: a luciferin-deficient, nonluminescent population located north of northern California and a luciferin-containing, luminescent population extending south of Cape Mendocino, California, to Baja California, Mexico. At the northern end of the southern population, a mixture of luminescent and nonluminescent P. notatus was found.     

Spectral composition of bioluminescence of epipelagic organisms from the Sargasso Sea

The spectral characteristics of single identified epipelagic sources of bioluminescence from the western Sargasso Sea were measured with an optical multichannel analyzer (OMA) system during the April, 1985, Biowatt cruise. The emission spectra of specimens representing 45 species from 8 phyla were measured. Peak bioluminescence emissions typically occurred between 440 and 500 nm, in the blue region of the visible spectrum. Three exceptions involved emission in the green, yellow, and red spectral regions. Intraspectific variability in spectra, was noted in several species. One shrimp species exhibited two modes of light emission, each with different emission spectra. Other cases involved dynamic color shifts of 10 to 14 nm; the source of the spectral variability is unknown, but may involve optical filtering or differences in the color of luminescence from multiple sites of light emission. Measurements from independent samples of unsorted plankton revealed different spectral distributions. This suggests that the spectral emissions of bioluminescence in the upper water column will vary, based on species assemblage.     

Comparative study of crustacean larval photoresponses

Ovigerous females of four brachyuran (Cancer gracilis, Lophopanopeus bellus bellus, Hemigrapsus oregonensis and Scyra acutiforns) and two anomuran (Pagurus beringanus and P. granosimanus) species, which live as adults in coastal areas, were collected near Friday Harbor, Washington, USA, in 1985, and spectral sensitivity and phototactic pattern of their larvae were measured. Responses were compared with previous measurements on estuarine species to determine whether responsiveness varies with adult habitat. Estuarine and coastal species have similar photoresponses. Spectral sensitivity of the test brachyran species had two maxima, one near 400 to 420 nm and another around 500 nm. The anomuran species were similar, but had an additional peak in the region of 580 to 620 nm. This sensitivity is adapted to daytime light conditions in the adult environment, and available spectra at the time of larval movement during diel vertical migration. Upon light adaptation and stimulation with a narrow light field, all six species showed positive phototaxis to high light intensities and a pronounced negative response to low intensities. Only the negative response will occur in natural underwater light conditions, and it is part of a predator-avoidance shadow-response which operates in areas of higher light intensity. The same phototactic pattern is observed in all species when darkadapted except H. oregonensis. If nocturnal vertical migration occurs, this negative response may be responsible for the descent at sunrise and depth maintenance during the day.     

Spectral sensitivity and visual pigment of the compound eye of the galatheid crab Pleuroncodes planipes

The compound eye of the galatheid crab Pleuroncodes planipes has been studied by means of visual pigment extractions and spectral sensitivity measurements under various conditions of adaptation. A photosensitive pigment with maximum absorption at 503 nm was identified. Its chromophore is retinal. The spectral sensitivity maximum is near 523 nm in the dark-adapted eye. Selective adaptation experiments suggest that the apical region of the eye has a homogeneous receptor population. The spectral sensitivity characteristics of P. planipes seem to be an adaptation to its mainly benthic life.     

Photosensitivity of the calanoid copepod Acartia tonsa

The light intensity and spectral sensitivities of the calanoid copepod Acartia tonsa Dana were determined by measuring phototactic responses. Adult females displayed only positive phototaxis. The dark-adapted copepod, which possesses a single naupliar eye, perceived light at intensities as low as 2.8x10¹¹ photons m^-2 s^-1. The action spectrum for positive phototaxis had no clear maxima but rather showed a broad range of greatest sensitivity from 453 to 620 nm. This sensitivity encompassed those wavelengths that are maximally available at the depth where the copepod is found during the day. This spectral overlap, coupled with the finding that the copepod requires light cues for nocturnal vertical miration, suggests that broad spectral sensitivity is an adaptive mechanism to maximize light intensity sensitivity during migration.     

Dinoflagellate luminescence increases susceptibility of zooplankton to teleost predation

The burglar alarm theory of bioluminescence was investigated by determining predation rates of a nocturnal teleost predator,Porichthys notatus, on nonluminescent kelp mysids illuminated by dinoflagellate flashes, between the fall and spring of 1989/1990. Mysids (Holmesimysis costata) were placed in aquaria containing varying concentrations (0 to 40 cells/ml) of the dinoflagellatePyrocystis fusiformis and a single midshipman fish. Controls usedP. fusiformis during their luminescence-inhibited day phase. Mysid swimming movements readily stimulated dinoflagellate luminescence. Flashes and prey strikes were observed simultaneously by image-intensifying and infrared video cameras on a splitscreen monitor. Predation rates increased at dinoflagellate concentrations of 3 to 15 cells/ml and decreased below controls at levels>20 cells/ml. Videotape analysis showed that at low concentrations (2 to 5 cells/ml), strike success rates exceeded 75% if prey were previously illuminated by a flash, but dropped below 50% at higher cell densities. Increased predation was attributed to luminescence revealing prey position. The decrease at higher concentrations was considered to be due to greater flash frequency providing a more diffuse and confusing target. The study demonstrates the effects of secondary luminescence on zooplankton predation at normally encountered dinoflagellate concentrations.     

Observations of planktonic bioluminescence in the euphotic zone of the California Current

The distributions of bioluminescence, temperature, salinity, oxygen. pH, and chlorophyll a were measured at 10 m intervals, to a depth of 100 m at a station (33°46N; 119°36W) in the California Current from 17 to 20 July 1982. The distribution of bioluminescence showed a marked day-night change which was consistent over the sampling period. The nighttime maximum was at the surface, and the daytime maximum was between 30 and 40 m. The shapes of the day and night distributions were independent of the absolute intensity of bioluminescence and were also insensitive to advection, as inferred from changing temperature-salinity relationships. The nighttime depth distribution broadened during a period of high wind Day to night differences in the color spectrum at the depth of maximum bioluminescence suggest that the luminescent organisms differed from day to night.     

Identification of marine organisms using kinetic and spectral properties of their bioluminescence

We have used a polychromator consisting of six photomultiplier tubes, each filtered to a different wavelength with narrow band-pass interference-filters, to study bioluminescence. Spectral and kinetic data collected from ten marine species in the laboratory describe their luminous flashes. These data suggest that the concept of luminous signatures, within the limits of our studies, is a valid one, with potential uses for future biological studies both in the laboratory and in situ. The kinetic parameters considered were rise time (RT), decay time (DT) and total time (TT), while the spectral parameters consisted of ratios of light intensities at 480 and 520 nm to the intensity at 500nm. Oneway analysis of variance (ANOVA) demonstrated significant heterogeneity among species for all variables. A posteriori analysis performed with the ANOVA for TT indicated that mean TT for most species is significantly different from all other species. Canonical discriminant analysis was performed to estimate the value of kinetic and spectral parameters for species' identification. Kinetic data were somewhat more valuable in species' classification than spectral data. Discriminant analysis with RT and DT alone gave 83.1% correct species-classifications. Classification success based only on relative intensities at 480 and 520 nm was 77.5%. When all four variables were included, classification success was 100%.     

Variability of spectral absorption efficiency within living cells of Pyrocystis lunula (Dinophyta)

This study examines variability in the spectral absorption efficiency in various parts of living Pyrocystis lunula cells. changes in spectral absorption efficiency measured within cells are attributed to light-induced chloroplast migration as well as reorganization of cellular material during the process of asexual reproduction. During the dark cycle, major pigment peaks were well resolved in those spectra measured in the distal cytoplasmic strands where chloroplasts were concentrated. In contrast, the absorption efficiencies measured in the granular central area that did not contain chloroplasts decreased gradually from the blue to the red portions of the spectrum and are similar to those published for detrital particles. When chloroplasts migrated toward the center of the cell in response to light, absorption efficiency curves for the granular central area were flatter than the curves measured in cytoplasmic strands containing chloroplasts. This was due to the combined absorption properties of the central area and the chloroplasts. Absorption efficiency spectra were also flattened in aplanospores within the parent vegetative cells because of the concentration of cellular material into smaller areas. These findings suggest that shapes of spectral absorption curves measured for the major phytoplankton groups cannot be assumed to remain constant over time. Furthermore, changes in cell structure may account for some of the reported diel changes in beam attenuation and stimulated fluorescence in natural waters.     

Planktonic bioluminescence in the pack ice and the marginal ice zone of the Beaufort Sea

An icebreaker cruise into the Beaufort Sea in the fall of 1986 provided a unique opportunity for studying planktonic bioluminescence in ice fields and in the marginal ice zone. Bathyphotometer casts (bioluminescence intensity, seawater temperature, beam attenuation coefficient, and salinity) and biological collections were made to a depth of 100 m. A light budget, which describes the planktonic species responsible for the measured bioluminescence, and a dinoflagellate species budget were constructed from the mean light output from luminescent plankton and plankton counts. The vertical distribution of bioluminescence among the ice stations was similar. The maximum intensities were 2 to 8×10⁶ photons s^-1 cm^-3 in the upper 50 m of the sea-ice interface. The marginal ice zone station (MIZ) exhibited a maximum intensity of 2 to 3×10⁸ photons s^-1 cm^-3 between 5 and 30 m depth. At Ice Station 2, Metridia longa and their nauplii contributed approximately 80% of stimulable bioluminescence in the upper 10 m but, overall, Protoperidinium spp. dinoflagellates contributed most of the light to a depth of 100 m. In the MIZ, Protoperidinium spp. dinoflagellates contributed 90% of the light within the upper 10 m, decreasing to 43% of the contributed light at a depth of 40 m. Below 40 m, dinoflagellate bioluminescence decreased to a few percent of the total to a depth of 90 m. Metridia spp. copepods contributed more than 50% of the light at depths from 40 to 90 m. Ostracods, larvaceans, and euphausiid furcilia contributed <1% of all bioluminescence at all depths sampled. Correlation analyses between measured bioluminescence (photons s^-1 cm^-3), the number of bioluminescent dinoflagellates and the light budget for the MIZ indicated highly significant associations: r=0.919, p=0.001, and r=0.912, p<0.001, respectively (Student's two-tailed t-tests). Bioluminescence was negatively correlated with seawater salinity at all stations (p=0.001). Maximum bioluminescence was measured in the less saline surface waters at all stations.     

Bioluminescence in the Monterey Submarine Canyon: image analysis of video recordings from a midwater submersible

Video images of bioluminescence were recorded in situ during a 1985 study of the midwater environment of the Monterey Canyon, using a single-person, untethered submersible. Gelatinous organisms were responsible for the most brilliant bioluminescent displays, often exhibiting elaborate kinetics in response to mechanical stimulation. Images of bioluminescent displays recorded from identified organisms are shown and display patterns are described. All bioluminescence emission spectra from captured specimens were blue, with peak emissions between 460 and 494 nm. Image-analysis of recordings of mechanically stimulated bioluminescence revealed source densities between 43 and 175 m^-3 and intensities between 2.5 and 37.3 W sr^-1 m^-3. The predominant display type at all depths studied (between 100 and 560 m) was luminous secretions. Despite high intensities of mechanically stimulated bioluminescence, no spontaneous light production was recorded in the absence of mechanical stimulation.     

Diel migration of phytoplankton and spectral light field in the Ría de Vigo (NW Spain)

Diel migration of Mesodinium rubrum, Eutreptiella sp., Scrippsiella trochoidea, Dinophysis acuminata and Ceratium furca throughout a 24 h cycle is described for a stable, well-stratified estuary (Ría de Vigo, NW Spain). Daily changes in light quantity and in spectral light ratios i.e. red:far-red, blue:red, green:red and blue:green have been analysed. The spectral light ratios changed at twilight and around noon at various depths. Some of the downward migrations were well predicted by Stokes' law, while other migrations were faster and deeper than calculated. The coincidence of these movements with abrupt changes in red:far-red, green:red and blue:green light ratios is discussed. Some species are able to migrate through the pycnocline, whereas others do not seem to be able to do so. Several species are present in maximum numbers at depth at night, while others display upward migration independent of light, suggesting the existence of endogenous rhythms. Upward migration at dusk began with dispersal of populations, with renewed aggregation at the sea surface coincident with an increase in the red:far-red ratio at 6 m and the green:red ratio at 6 and 10 m. Based on direct evidence for the control of flagellar mobility by light quality reported by other authors from laboratory studies, it is suggested that, together with other cues, spectral light ratios of different light qualities modulate vertical phytoplanktonic migration. Received: 18 July 1997 / Accepted: 17 October 1997     

Luminescence in the oxidation of isoproterenol by the superoxide anion radical in dimethyl sulfoxide

Chemiluminescence appearing during oxidation of isoproterenol using chemical system involving superoxide anion radical has been studied. Chemiluminescence and fluorescence spectra were measured. The chemiluminescence spectrum was measured with cut‐off filters and revealed bands with maximum at 440, 480, 550, 640 and 700 nm. The bands at 480, 640 and 700 nm were similar to those observed for singlet oxygen. The fluorescence spectrum exhibited maximum at 560 nm.

The inhibitory effect of several biologically important compounds known as O₂ ^?‐ HO^? and ¹O₂ scavengers on the light emission was studied.

The obtained results indicate that oxidation of isoproterenol by O₂ ^?‐ involves products in the electronically excited states. The data also seem to indicate the protective effect of isoproterenol on the deoxyribose degradation.     

Comparative study of behavioral-sensitivity thresholds to near-UV and blue-green light in deep-sea crustaceans

The behavioral sensitivities of five species of deep-sea crustaceans (order Decapoda: Acanthephyra curtirostris, A. smithi, Notostomus gibbosus, Janicella spinacauda and Oplophorus gracilirostris) to near-UV and blue-green light were studied during a research cruise off the coast of Hawaii in 1993. Two of the five species have electrophysiologically-measured spectral sensitivity peaks at 400 and 500 nm, while the remaining three species have a single sensitivity peak at 490 to 500 nm. In the current study, behavioral mean threshold sensitivities (defined as the lowest irradiance change to which the shrimp would give a behavioral response) were determined for tethered specimens of each species at two wavelengths, 400 and 500 nm. The mean behavioral threshold sensitivities of the two species with putative dual visual-pigment systems were approximately the same to near-UV and blue-green light, while the other three species were significantly less sensitive to near-UV vs blue-green light. Results from these experiments indicate that (1) behavioral information obtained from tethered shrimp accurately reflects their spectral sensitivity, and (2) the sensitivity of the putative dichromats to near-UV light is sufficiently low to detect calculated levels of near-UV light remaining in the down-welling field at their daytime depth of 600 m. Possible functions of this high sensitivity to short wavelength light are discussed.     

An unusual blue mesogleal protein from the mangrove jellyfish Cassiopea xamachana

The jellyfish Cassiopea xamachana often contains a blue pigment diffused within the acellular portion of its masoglea. In the bell, both the pigment and endosymbiotic zooxanthellae are concentrated immediately beneath the ex-and subumbrellar epithelia. Chromatographic and polyacrylamide gel electrophoretic techniques demonstrate that the pigment is a highly polymeric glycoprotein (mol. wt>10⁶ daltons) comprised of two subunits with molecular weights of 34 500 and 30 300 daltons and characterized by multiple charged species. The blue native protein exhibits light absorption maxima at 620, 587, 555 and 415 nm, while SDS denatured protein is pink with a single absorption maximum at 507 nm. No prosthetic chromophore or heavy metal component was detected. The pigment is proposed to act as a light attenuator protecting the jellyfish from injurious solar irradiation while allowing photosynthetically active wavelengths to reach the zooxanthellae.     

Some features of the bioluminescence of the radiolarianThalassicolla sp.

The luminescence of the solitary radiolarianThalassicolla sp. has a spectral emission maximum at 446±4 nm and the luminescence of crude homogenates can be activated by calcium ions. Electrical stimulation of single individuals frequently causes them to produce repetitive post-stimulus flashing akin to that reported in a number of other marine animals. The possible relationships between the luminescent system ofThalassicolla sp. and of certain coelenterates are discussed.