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.     

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     

Eyes and extraocular photoreceptors in midwater cephalopods and fishes: Their roles in detecting downwelling light for counterillumination

The means of detecting downwelling light for counterillumination in several midwater animals has been examined. Eyes and extraocular photoreceptors (drosal photosensitive vesicles in the enoploteuthid squid Abraliopsis sp. B and pineal organs in the myctophid fish Myctophum spinosum) were alternately exposed to overhead light or covered by a small opaque shield above the animal and the bioluminescent response of the animal was monitored. Covering either the eyes or the extraocular photoreceptors resulted in a reduction in the intensity of counterillumination. Preliminary experiments examining the bioluminescent feedback mechanism for monitoring intensity of bioluminescence during counterillumination in the midwater squid Abralia trigonura indicated that the ventral photosensitive vesicles are responsible for bioluminescent feedback.     

Photopigment and spectral sensitivity in the bioluminescent fish Porichthys notatus

Digitonin extracts from retinas of the marine fish Porichthys notatus, from California and Puget Sound (USA), contain a single photopigment with maximum absorbance at 498 nm. In the presence of hydroxylamine the final product of bleaching absorbs maximally at 367 nm, indicating that the photopigment is based on retinal. Spectral sensitivity measurements performed on intact eyes and eye-cup preparations in darkadapted and selectively light-adapted states yield curves which suggest that the retina of P. notatus contains at least 3 different types of receptors. In the darkadapted state, the spectral sensitivity curve has a broad peak, with a maximum between 480 and 500 nm. In the blue and red light-adapted states, in addition to depressing the sensitivity, the shape of the sensitivity curve also changes. Under these conditions new peaks appear which suggest the presence of at least two different types of cones, with sensitivity maxima in the blue and the green regions of the spectrum, respectively. Comparison of the spectral sensitivity curves with the in vivo bioluminescence emission spectrum of the fish gives an especially good correspondence and suggests that the fish is able to perceive the bioluminescence of its own species.     

Counterillumination in the Hawaiian bobtail squid,<Emphasis Type="Italic"> Euprymna scolopes</Emphasis> Berry (Mollusca: Cephalopoda)

The mutualism between the Hawaiian bobtail squid Euprymna scolopes and the luminescent symbiont Vibrio fischeri has been used extensively as a model system for studies ranging from co-speciation and biogeography to gene regulation and the evolution of pathogenesis. In this association, the luminescent bacterium V. fischeri is housed in a complex light organ within the mantle cavity of E. scolopes. Prior hypotheses have assumed that sepiolid squids in general utilize the bioluminescence produced by their V. fischeri symbionts for counterillumination, a behavior that helps squid camouflage themselves by matching down-welling moonlight via silhouette reduction. This assumption, based solely on the morphology of the squid light organ, has never been empirically tested for Euprymna in the laboratory. Here, we present data demonstrating that E. scolopes can modify the intensity of light produced by V. fischeri in the light organ as down-welling light intensity changes. Bacterial bioluminescence from the light organ is directly correlated with down-welling light intensity, suggesting that E. scolopes individuals utilize and control V. fischeri luminescence for counterillumination.Communicated by P.W. Sammarco, Chauvin     

Population interconnectivity and implications for recovery of a species of concern, the Pacific hake of Georgia Basin

To aid the recovery of a species, understanding the extent to which populations are connected is useful for targeting conservation efforts. Pacific hake within waters of Puget Sound, Washington State, USA, and Georgia Strait, British Columbia, Canada are listed as a species of concern under the U.S. Endangered Species Act due to dramatic declines in the Puget Sound population. To assess the role of dispersal in the recovery of Pacific hake, we sought to quantify patterns of connectivity between populations in Puget Sound and Georgia Strait. Using natural chemical markers from otoliths of fish sampled from these two populations, we linked natal signatures of fish to signatures of individuals from known spawning grounds. Results indicated that 82 % of individuals collected from Puget Sound (n = 78) were estimated to have originated there, while 40 and 92 % of the individuals collected from two cohorts within Georgia Strait (n = 9 and 24, respectively) had originated from Puget Sound. A trend of “population abandonment” of fish from Puget Sound suggests that recovery of this Pacific hake population will depend on local management practices.     

Inception of formation and early morphogenesis of the bacterial light organ of the sea urchin cardinalfish, <Emphasis Type="Italic">Siphamia versicolor</Emphasis>

The cardinalfish Siphamia versicolor (Perciformes: Apogonidae) forms a bioluminescent symbiosis with the marine luminous bacterium Photobacterium mandapamensis, harboring the bacteria in a ventral, disc-shaped light organ and using the bacterial light apparently for counterillumination and attracting prey. Little definitive information has been available on the developmental and microbiological events surrounding the initiation of symbiosis, a critical stage in the life history of the fish, in S. versicolor or any of the many other species of bacterially luminous fish. To identify the stage at which light organ formation begins, to determine the origin of cells forming the light organ, and to characterize its bacterial colonization status during development, early developmental stages of S. versicolor obtained and reared from wild-caught mouth-brooding males were examined with histological and microbiological methods. A light organ primordium was not evident in embryos, post-embryos, or pre-release larvae, whereas the light organ began to form within 1 day of release of full-term pre-flexion larvae from the mouths of male fish. Analysis of post-release larvae revealed that the light organ arises from a proliferation and differentiation of intestinal epithelial cells, and that it quickly develops structural complexity, including the formation of chambers and gaps contiguous with the intestinal epithelium. However, the nascent light organ remained uncolonized by the symbiotic bacteria through several days of post-release development, even in the presence of high numbers of the symbiotic bacteria. These results demonstrate that the inception of light organ formation in S. versicolor occurs independently of its symbiotic bacteria and that receptivity to bacterial colonization apparently requires substantial post-release development of the light organ. Larvae therefore most likely acquire their symbiotic bacteria from seawater, during or shortly after the transition from the pre-flexion to the flexion developmental stage.     

Presence of coelenterazine in mesopelagic fishes from the Strait of Messina

Coelenterazine, a luciferine, and luciferase activity specific to coelenterazine were detected and assayed in various tissues of mid-water fishes, Argyropelecus hemigymnus, Chauliodus sloani, Myctophum punctatum, Vinciguerria attenuata and Cyclothone braueri. Coelenterazine was found mainly in digestive systems and photophores of these fishes. A large species to species variation was found: extremely high levels of coelenterazine and luciferase were found in the M. punctatum digestive system whereas only a very low level of coelenterazine was detected in C. braueri. Coelenterazine was detected in A. hemigymnus eggs supporting the hypothesis of a maternal transfer of luminous capabilities. Luciferase activity specific to coelenterazine was found in photophores as well as in various other tissues suggesting another (besides light emission) biological function for this enzymatic activity. Distribution of coelenterazine in all tissues of the individuals supports the hypothesis of the dietary acquisition of coelenterazine by these fishes.     

Evidence for multiple recruitment-cohorts of Puget Sound Dungeness crab,Cancer magister

While sampling intertidally in Puget Sound, Washington, USA, for juvenile Dungeness crab (Cancer magister) in 1984, we found evidence of two distinct cohorts of the same year-class based on sizes of first-instar juveniles (J1) and the spatial/temporal patterns of settlement. In 1988, three distinct cohorts were observed to settle in Puget Sound and its approaches. Settlement of one cohort occurred during May in the Strait of Juan de Fuca and in those areas of Puget Sound closest to the Strait. J1 individuals of this cohort were large (x=7.4 mm carapace width, CW) and comparable in both size and timing of settlement to populations along the Washington coast (e.g. Grays Harbor and Willapa Bay). Initial settlement density of the May cohort was as high as 215 crabs/m² in intertidal eelgrass beds along the Strait of Juan de Fuca and decreased to <2 crabs/m² within Puget Sound and the Strait of Georgia. A second cohort apparently originated in Hood Canal (a deep inland fjord), its size upon settlement in June was significantly smaller (J1 x= 5.3 mm CW) than the May cohort, and it was limited to Hood Canal and areas of Puget Sound close to the mouth of Hood Canal. A third cohort, which settled in late July and August, was the smallest of the three cohorts (J1 x= 4.8 mm CW), and was widely distributed around Puget Sound from Seattle in the south to the USA/Canadian border in the north. We hypothesize that most juvenile recruitment in Hood Canal and Puget Sound originates from parental stocks endemic to their respective basins (Hood Canal and Puget Sound cohorts), but that, on occasion, oceanographic conditions allow substantial influx of Pacific Ocean Dungeness crab larvae (oceanic cohort) through the Strait of Juan de Fuca into Puget Sound. Tracking of spatial/temporal settlement patterns and comparison of J1 sizes proved useful for estimating the probable sources and dispersion of Dungeness crab larvae. Differences in size and time of settlement between various larval cohorts of C. magister may prove useful as biomarkers for tracing circulation patterns within and between inland waters of Washington and the Pacific Ocean. Causes of smaller size and later settlement of the Puget Sound cohort relative to oceanic conspecifics of the same year-class are discussed.Contribution No. 856, School of Fisheries, University of Washington, Seattle     

Three new modes of luminescence in the leiognathid fish Gazza minuta: Discrete projected luminescence,ventral body flash,and buccal luminescence

Three new modes of luminescence are described for Gazza minuta (Bloch) (Perciformes: Leiognathidae) as observed in specimens collected in the Philippines in April and May, 1982: discrete projected luminescence (DPL), ventral body flash, and buccal luminescence. DPL sharply contrasts with previously reported modes of diffuse luminescence in leiognathids (counterillumination and opercular flash) in being a pair of bright collimated beams of light emanating from the fish in an anteroventral direction. The brightness, coherence, directionality, and control of DPL suggest striking similarities to luminescence in anomalopid (flashlight) and monocentrid (pinecone) fishes and perhaps in certain apogonids (cardinalfishes). The structural correlate for DPL is a small clear patch of skin lying at the posterior margin of each opercular cavity. Luminescence from the internally located light organ traverses transparent bone and translucent muscle before passing through the clear skin of the patch area. Behavioral and anatomical observations of ventral body flash and buccal luminescence are also presented. These new modes of luminescence indicate a much greater than expected diversity of luminescent behaviors in leiognathids, perhaps greater than that of any other organism yet studied. The internal location of the light organ is recognised as providing the potential for this diversity.     

Zinc and cadmium in the Pacific hake Merluccius productus off the western U.S. coast

Concentrations of zinc and cadmium in muscle tissue or whole Pacific hake, Merluccius productus (Ayres), increase with increasing total wet weight, especially for smaller fish. These trends appear to be related to the euphausiid diet of hake. Except for the isolated population in Puget Sound, no relationship between zinc or cadmium concentration and collection site is apparent.     

Transfer of symbiotic luminous bacteria from parental Leiognathus nuchalis to their offspring

Juveniles of Leiognathus nuchalis were raised from fertilized eggs for up to 60 d and examined for luminescence activity. Almost all juveniles raised separately from adults failed to produce detectable light. In contrast, a significant percentage (33 to 100%) of the juveniles became luminescent in less than 48 h when they were either kept with adults or inoculated with a homogenate of the adult light organs. The luminescence tended to increase with time after the treatments. These findings suggest that: (1) most of L. nuchalis offspring typically hatch and develop apo-symbiotically and (2) at least 45 d after hatching, juveniles can be infected with symbiotic luminous bacteria from the light organ of adult fish, and thereby gain the ability to produce light. Received: 16 February 1999 / Accepted: 21 September 1999     

Red bioluminescence in fishes: on the suborbital photophores of Malacosteus,Pachystomias and Aristostomias

Many deep-sea animals produce blue bioluminescence, but species of three genera of midwater dragonfishes also produce red light. In addition to numerous small body photophores, species of these genera (Malacosteus, Pachystomias and Aristostomias) have large suborbital photophores that emit red light and postorbital ones that emit blue light. Pachystomias microdon additionally has a red-emitting preorbital photophore. Fluorescence measurements from the intact suborbital photophores, and from their exposed cores, confirm the previous hypothesis that the red light emitted by Malacosteus is spectrally altered by a superficial shortwave cutoff brown filter. No such filter is present in the other two genera. Studies of the anatomy of the photophores show that the suborbital photophore of Malacosteus is similar in general organisation to other photophores, including the postorbital photophore. The red-emitting photophores of Pachystomias and Aristostomias, however, have a unique organisation, in which the bulk of the photophore comprises a gland that lies outside the thick reflector. Strands of tissue run from the gland into the photogenic core of the photophore through numerous pores in the reflector. The production and use of red light by these fishes is discussed in the context of these results.Electronic Supplementary Material Supplementary material is available for this article at     

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.     

Differences in morphology and habitat use among the native mussel <Emphasis Type="Italic">Mytilus trossulus</Emphasis>, the non-native <Emphasis Type="Italic">M. galloprovincialis,</Emphasis> and their hybrids in Puget Sound,Washington

Mytilus galloprovincialis (Mg), the Mediterranean blue mussel, is sympatric with the native M. trossulus (Mt) throughout much of the north Pacific, likely as the result of human introduction. We investigated the distribution of the two species and their hybrids (Mgt) in Puget Sound, Washington, to determine whether differences occur in habitat preference between the two species and hybrids. In addition, we investigated whether there were consistent size and shape differences between the native and introduced mussels and hybrids. Measurements of over 6,000 mussels from 30 sites, of which 1,460 were genotyped for a species-specific genetic marker, revealed that Mg and Mgt can be found throughout Puget Sound. Mg and Mgt were larger and exhibited a greater height:length ratio than Mt. Frequencies of Mg and Mgt were higher in subtidal habitats, such as docks, than on intertidal rocks, walls or pilings. Within intertidal habitats, Mg and Mgt were more frequent than Mt in the lower reaches of the intertidal. At slightly more than half the sites the frequency of the three genotypes accorded with random mating expectations suggesting no consistent barriers to gene flow between species. The standardized random sampling methods and simple morphometric identification techniques described here can be used to test whether the frequency of invasive mussels changes over time and space in Puget Sound.     

Dynamics of benthic vegetation standing-stock,irradiance, and water properties in central Puget Sound

The standing stock of benthic macroalgae, sediment-associated microalgae and eelgrass (Zostera marina L.) was sampled in conjunction with irradiance and water properties from June 1982 through March 1984 to examine the relationship between the dynamics of benthic primary producers and environmental factors in central Puget Sound, Washington, USA. Sediment-associated microalgal standing stock (measured by chlorophylla) peaked in April and August. The seaweed assemblage, dominated by bladed green algae (e.g.Ulva fenestrata) and eelgrass exhibited maximum standing stocks in August. Although water temperature correlated best with changes in standing stock of all vegetation types, solar irradiance appeared to trigger the onset of biomass buildup and autumn die-back by the plants. Seasonal variations in dissolved oxygen reflected the buildup and loss of plant standing-stock. Nutrient concentrations, with the exception of ammonia, exhibited seasonal trends. Most nutrients were in greatest concentration in winter and reached minimum concentration in late spring-summer. Regeneration of nutrients in autumn followed shortly after the autumn dieback of the benthic vegetation. We concluded that irradiance was the primary controlling factor in the system. Nutrient limitation (primarily nitrate) may control standing-stock accumulations from the period May–October when light is not limiting. In contrast to phytoplankton systems in deep portions of Puget Sound, shallow nearshore systems may be more susceptible to the effects of increased inorganic nutrient-loadings from anthropogenic sources.Contribution No. 808, School of Fisheries, University of Washington     

Luminescence from non-bioluminescent tissues in oceanic cephalopods

Several tissues (e.g. kidney, blood, digestive gland) in oceanic cephalopods which do not exhibit in vivo bioluminescence, luminesce when homogenized in the presence of air or when simply exposed to air in a vial (blood). The source of the luminescence appears to be a luciferin: treatment of kidney homogenates and blood with a photophore extract presumably containing luciferase resulted in a 20-fold increase in light production. Luminescence was also found in the renal fluid, which may be the source of luminescent clouds produced by squids. The variability in luminescence found in some tissues of cephalopods appeared to be related to feeding. Luminescence was also detected in the digestive glands of midwater octopods.     

Population genetics and systematics of theLeptasterias hexactis (Echinodermata: Asteroidea) species complex

Morphological characters and 14 enzyme loci were examined for 1 040 sea stars, currently recognized as forms ofLeptasterias hexactis, from Lynn Canal, Alaska, and Puget Sound, Washington, USA, between March 1988 and April 1989. Three morphologically and two genetically distinctLeptasterias forms were identified. The most common form found at both localities wasL. epichlora (Brandt)sensu Verrill.L. hexactis (Stimpson)sensu Verrill co-occurred withL. epichlora at all study sites and apparently hybridizes extensively withL. epichlora in the Puget Sound region, but rarely, if at all, in Alaska. The presumptive product of this hybridization morphologically resembledL. aequalis (Stimpson)sensu Fisher, and was conspicuously absent from Alaskan samples. Considerable genetic distance existed betweenL. epichlora andL. hexactis (Nei'sD=0.19±0.01) and moderate genetic differentiation occurred between populations of each species from Alaska and Washington (Weir and Cockerham'sF _RT =0.29±0.04 forL. epichlora and 0.21±0.15 forL. hexactis). A significant (p<0.05) deficiency in the proportion of heterozygous individuals was found compared to Hardy-Weinberg expectations (Wright's fixation index,F _ID=0.12±0.04 and 0.31±0.08 forL. epichlora andL. hexactis, respectively). However, mean observed heterozygosity for each species (0.09±0.03, 0.14±0.04 and 0.14±0.04 forL. epichlora, L. hexactis andL. aequalis, respectively) fell within the range of reported values for other asteroid species (ca. 0.04 to 0.37). The results of this study indicate that considerable genetic integrity is maintained betweenL. epichlora andL. hexactis, which warrants their recognition as distinct species despite their apparent hybridization in the Puget Sound region.     

The dynamics of phytoplankton blooms in puget sound a fjord in the Northwestern United States

This paper describes a quantitative investigation of relationships between the growth of phytoplankton, and climatic and hydrodynamci conditions in temperate fjords with marked tides, as exemplified by Puget Sound, Washington (USA). Algal growth in the open waters of the central basin of the Sound is dominated by a number of intense blooms beginning in late April or May and recurring throughout the summer. Rarely, and only briefly, does nitrate become exhausted. The phytoplankton production rate in the central basin of Puget Sound is about 465 g C m^-2 year^-1. During the springs of 1966 and 1967, oceanographic measurements were carried out at a mid-channel station with sufficient frequency to allow investigation of physical and biological processes with time scales of the order of a day. The principal investigative tool is a numerical model in which the hydrodynamical conditions are represented by an approximate analysis of the gravitational convection mode of circulation. Algal concentration is represented as a continous function of space and time in the model which ascribes changes in phytoplankton density to variations in photosynthetic and respiratory activity, algal sinking, grazing by herbivores, and to mixing and advection. Computations adequately reproduce the principal features of phytoplankton concentrations observed during 75 days and 35 days in the springs of 1966 and 1967, respectively. Numerical experiments assess the relative importance of various processes which govern the level of primary production in Puget Sound. It is concluded that phytoplankton growth is limited by a combination of factors, including vertical advection and turbulence, modulation of underwater light intensity by self-shading and inorganic particulates, sinking of algal cells, and occasional rapid horizontal advection of the population from the area by sustained winds. The high primary productivity of the Sound is due to intensive upward transport of nitrate by the estuarine mechanism. These results should be generally applicable to other temperate fjords because of the largely conventional choice of the biological functions.     

Larval development of the crabCancer magister in temperature regimes simulating outer-coast and inland-water habitats

Postlarval megalopae of the Dungeness crabCancer magister inhabiting offshore coastal waters along the west toast of North America are larger and settle earlier than do those occupying the inland waters of the Puget Sound basin (Washington, USA, and British Columbia, Canada). The Puget Sound habitat is characterized by low initial temperatures that steadily increase during the course of zoeal development, while offshore temperatures are more moderate and stable. Larvae were raised in the laboratory from hatching to megalopa in three temperature treatments: (1) constant 10 °C (CO), (2) a regime that temporally simulated temperatures found off the central California coast during the larval season (CA); and (3) a regime simulating temperatures found within the Puget Sound basin (PS). Zoeal duration was 44% longer in the PS treatment than in the other two, at least partially accounting for observed differentes between outer-toast and inland-water settlement times. Although differentes were measured in megalopal weights and carapace lengths among treatments, results do not explain differences in megalopal size observed between outer-toast and inland-water individuals. Survival to megalopa was highest in the PS treatment, with the difference due to significantly lower mortality than in CO and CA treatments during the terminal zoeal stage. Daily instantaneous mortality rates were lower in the PS than in CO or CA treatments, indicating Chat extended larval duration will not necessarily result in reduced settlement success.