Control of bloom in a tropical lake: grazing efficiency of some herbivorous fishes

Using a limnocorral study, Microcystis grazing by silver carp, bighead and tilapia was examined in an eutrophic shallow lake permanently infested with a Microcystis bloom. Twelve fingerlings of each fish species were introduced into the limnocorrals placed in the lake, in triplicate, and their grazing effects were examined over a period until the limnocorral bloom declined substantially and remained unchanged. Introduction of fish, regardless of species, into the limnocorrals resulted in sharp reductions (60–93%) of the initial Microcystis population on day 3 or 7. Maximum gross and net clearance of Microcystis were by silver carp followed by bighead and tilapia. Short-term high retention of Microcystis by tilapia indicated that the clearance effect did not last long due to a high rate of defaecation of undigested Microcystis . Nearly 6–180% nutrient enrichment of the limnocorrals was attributable to the defaecation of test fishes suggesting their ichthyoeutrophication potentials, which were in the following order: tilapia>bighead>silver carp. It is concluded that silver carp is most suitable for clearing Microcystis in the lake because of its minimal ichthyoeutrophication effect.     

Control mechanisms of arctic lake ecosystems: a limnocorral experiment

To assess the potential impact of human exploitation on arctic lakes and to determine how these eco systems are regulated we initated a limnocorral experiment in Toolik Lake, Alaska, in the summer of 1983. The limnocorrals were 5 m in diameter and from 5–6 m in depth and were open to the sediments. In 1983 four limnocorrals were deployed in an isolated bay of Toolik Lake within a cross-classified treatment regime of high and low inorganic nitrogen and phosphorus additions and high and low free swimming fish additions. The objective of the nutrient addition was to stimulate phytoplankton growth and determine the extent to which increased plant production was passed through pelagic and benthic food chains. The objective of the fish addition was to determine the impact of fish predation on large-bodied zooplankton, especially the zooplanktivorous copepod Heterocope, then to study the effect of altered Heterocope densities on small-bodied zooplankton species population dynamics. In 1984 two more limnocorrals were deployed, one a low fish, 1 × nutrient addition treatment and the other a no fish, no nutrient treatment. The fish manipulation was changed to confining several fish in cages with the cages held in corrals for varying lengths of time. The addition of inorganic nitrogen and phosphorus dramatically increased phytoplankton productivity. This increase in algal biomass and production greatly altered the light environment and water quality in the nutrient treated limnocorrals. The secchi disk depth in the nutrient treated limnocorrals declined each summer reaching as low as 1 m in 1985. Both oxygen content and pH increased in the nutrient treatment corrals. Corrals not receiving nutrient additions remained near lake concentrations for most water quality parameters. While phytoplankton biomass was stimulated in 1983 phytoplankton growth was not sufficient to draw down all the nitrogen and phosphorus added and these nutrients reached high levels in the last half of the summer. In 1984 phosphorus remained above 20 μg in the nutrient-treated corrals but ammonia dropped to reference levels by day 25. In 1985 both nutrient concentrations rapidly declined to reference levels. Most pelagic components responded to the nutrient additions. Microbial production was stimulated in the nutrient treated limnocorrals and bacterial population sizes built up to nearly 8–10 times those of the reference corrals. However, microheterotrophs soon increased in abundance and apparently grazed down bacteria to reference levels. Phytoplankton population density, as estimated by chlorophyll a determinations, increased dramatically with nutrient addition such that each year the phytoplankton densities were higher than before. Primary productivity was also stimulated and appeared not to be light limited even when phytoplankton densities rose to high levels. In the first two years of the experiment zooplankton densities were little altered by the increased phytoplankton densities. However, by 1985 daphnid densities were quite a bit higher in the high nutrient addition limnocorrals. The benthic community and sediment response was much less affected by nutrient addition. Overall sediment respiration increased in the nutrient treated corrals but underlying sediments seemed little affected. Decomposition of Carex litter was likewise little affected by nutrient addition. Benthic invertebrates were also little impacted by the nutrient addition and increased sedimentation of phytoplankton. However, the response of benthic invertebrates is difficult to assess fully in the current experiment because chironomids, a prominent component of the benthic community, failed to recruit into the limnocorrals and the corrals physically shifted during ice-out in the spring of 1984 disturbing the sediment in several corrals. The fish additions in 1983 of free swimming grayling essentially eliminated large bodied zooplankton, especially Heterocope septentrionalis, from all four limnocorrals. In subsequent summers Heterocope were not so dramatically preyed upon but generally were found in higher densities in the low or no fish treatments. However, either when Heterocope were eliminated in 1983 or were in rough inverse proportion to fish density, altered Heterocope abundance had no obvious affect on small-bodied zooplankton abundance. The fish treatment apparently influenced the zooplankton response to high nutrient addition in 1985. In the high nutrient limnocorrals daphnid populations became very abundant, but in the high fish treatment the daphnid responding was the small-bodied D. longiremis while in the low fish treatment the daphnid responding was the large-bodied D. middendorffiana. Thus we have considerable evidence for bottom up control of phytoplankton density and production. This increased production ultimately, but not for two years, stimulated zooplankton density increases. Increased nutrients had little effect on the benthos or sediments. Fish manipulations influenced large-bodied zooplankton but had little effect on small-bodied zooplankton. Because grayling are predominantly plankton feeders in lakes, no fish effect on benthic invertebrates was expected. Limnocorrals thus seem good systems to study nutrient-phytoplankton interactions. They are not as suitable for benthic invertebrate studies and fish manipulations may be difficult. Most other limnocorral studies were of brief duration; however, in the present study the limnocorrals seemed to perform well over a three year period.     

An inexpensive, deep-water limnocorral that compensates for wave action

SUMMARY. 1. A large reusable limnocorral is described, suitable for use in water bodies with substantial wave action. The clear plastic enclosure is 1 m in diameter, 25m deep, lightweight (∼50 kg), and inexpensive ($350 U.S.A.).
2. The lower portion of the limnocorral is designed to hang motionless below the water surface, while an expandable floating collar section moves freely up and down with any wave action.
3. Two limnocorrals, equipped with removable sedimentation traps at the bottom, were field tested successfully for 6 weeks in Flathead Lake, Montana, withstanding wave heights up to 1m. Chlorophyll levels and profiles of various physical-chemical parameters measured inside the limnocorrals were comparable to Flathead Lake values.
4. Despite their large size, the limnocorrals are relatively easy to transport, install and retrieve.     

Cascading trophic interactions: Uncoupling at the zooplanktonphytoplankton link

Four limnocorrals (8 m dia. by 15 m deep) located at Lake St. George, Ontario, Canada; were used to examine the interactions between planktivorous fish, crustacean zooplankton (notably Daphnia galeata mendotae), and phytoplankton. During the spring, in all of the limnocorrals, high Daphnia biomasses were correlated with increased water transparency and a spring clear-water phase. However, as the summer progressed, the relationship between Daphnia biomass and phytoplankton abundance became more complex and less predictable. Investigation of these interactions suggested four conclusions. (1) During late July and throughout August and September, water transparency decreased and algal cell counts increased. In 3 of 4 limnocorrals, deterioration in water quality occurred 3–5 weeks before zooplankton (and Daphnia populations) declined. In all cases decreased transparency was associated with increased concentrations of algal cells (Gloeococcus) that were poor food sources for Daphnia. These results suggested that decreased water transparency was not caused by decreases in Daphnia biomass. (2) Taken together, data from all of the limnocorrals showed no correlation between the magnitude of July Daphnia biomasses and the percentage of poor food source algae that were observed in August. This suggested that grazer effects were not necessary for the onset of summer poor food source algal blooms. (3) In two limnocorrals, there was a positive correlation between increased Daphnia mortality and the onset of poor food source blooms. In the other two limnocorrals there was no correlation. In all limnocorrals there was no correlation between decreased Daphnia reproductive capacity and poor food source blooms. These data suggested that blooms of poor food source algae were not necessary for the collapse of Daphnia populations. (4) In all 4 limnocorrals there was a strong correlation between the time that 0+ yellow perch planktivores reached biomasses of 30–50 kg ha^–1 and the collapse of Daphnia populations Species and size selection was also observed. These results suggested that for this set of limnocorral experiments, fish biomasses in the 30–50 kg ha^–1 were responsible for the collapse of Daphnia populations in the summer.     

Utilization of prawn waste for chitinase production by the marine fungus Beauveria bassiana by solid state fermentation

Prawn waste, a chitinous solid waste of the shellfish processing industry, was used as a substrate for chitinase production by the marine fungus Beauveria bassiana BTMF S10, in a solid state fermentation (SSF) culture. Theprocess parameters influencing SSF were optimized. A maximum chitinase yield of 248.0 units/g initial dry substrate (U/gIDS) was obtained in a medium containing a 5:1 ratio (w/v) of prawn waste/sea water, 1% (w/w) NaCl,2.5% (w/w) KH2PO4, 425–600m substrate particle size at 27°C, initial pH 9.5, and after 5 days of incubation. The presence of yeast extract reduced chitinase yield. The results indicate scope for the utilization of shellfish processing (prawn) waste for the industrial production of chitinase by using solid state fermentation.     

The effects of dissolved macro-nutrients on the herbaceous vegetation around dune pools

In Dutch coastal dunes the relation was studied between the composition of the vegetation on the banks of pools and the nutrients supplied by the groundwater. Areas infiltrated with eutrophic river water were compared with non-affected areas. The floristic composition was interpreted according to classification systems of ruderal, nitrophilous tall hemicryptophytes and the conservation parameters richness and rarity. Of the three main macro-nutrients (orthophosphate, nitrate and potassium), the orthophosphate load shows the highest correlation with the vegetation variables. It is suggested that a reduction of orthophosphate during pre-purification of the infiltration water would reduce the dominance of the ruderals to the extent that the original dune slack species could return locally.Nomenclature of species follows Arnolds & Van der Meijden (1975).We thank G. J. de Bruyn, W. H. van Dobben, W. T. de Groot, W. J. ter Keurs, F. van der Meulen, W. H. van der Molen, M. A. W. Noordervliet, and I. S. Zonneveld for their comments on the study; J. Braak, J. Brakkee and R. van Nood for their assistance with the chemical analyses; A. R. Kaal for the translation, and G. P. G. Hock for the drawing of the figures.     

Discrimination of objects through electrolocation in the weakly electric fish,Gnathonemus petersii

Summary Three weakly electric fish (Gnathonemus petersii) were force-choice trained in a two-alternative procedure to discriminate between objects differing in their electrical characteristics. The objects were carbon dipoles in plexiglass tubing (length 2.5 cm, diameter 0.6 cm). Their electrical characteristics could be changed by varying the impedance of an external circuit to which they were connected (Fig. 1). In one (the capacitance dipole) the resistance was very low(< 3 ) and the capcitance variable. In the other (the resistance dipole) the resistance was variable and the capacitance low (<50 pF).Capacitances from several hundred pF (lower thresholds, Fig. 2) to several hundred nF (upper thresholds, Fig. 3) could be discriminated from both insulators and good conductors. In all cases the reward-negative stimulus was the capacitance dipole, which was avoided by all fish spontaneously. Thresholds were defined at 70% correct choices.The fish were then tested for their ability to discriminate between one object with a given capacitance and another with resistances varying from 3 to 200 k. The capacitance dipole continued to be the negative stimulus throughout. All 3 fish avoided it in at least 80% of the trials at each stimulus combination (Fig. 4). This result suggests that Gnathonemus perceives the capacitance and the resistance of objects differentially.The effect of the dipole-objects as well as some natural objects on the local EOD was recorded differentially very close to the fish's skin (Fig. 5). The amplitude of the local EODs was affected by all types of objects as they approached the skin. However, the waveform was changed only by capacitance dipoles and some natural objects (Figs. 6 and 7). It appears that the fish perceive not only intensity changes in the local EOD but wave-form deformations as well and can thus distinguish objects of different complex impedances.Abbreviations EOD electric organ discharge - f _max maximal spectral frequency - GP Gnathonemus petersii - LFS local filtered signal - PMA probing motor act - S+ positive stimulus - S negative stimulus     

Comparative efficiencies of bispecific F(ab′γ)2 and chimeric mouse/human IgG antibodies in recruiting cellular effectors for cytotoxicity via Fcγ receptors

Summary The three forms of Fc receptor carried by monocytes (FcRI, II) and natural killer (NK) cells (FcRIII) are all capable of mediating cell lysis. Here we compare the use of F(ab)₂ bispecific antibodies, specifically targetting individual FcR, and chimeric IgG mouse/human antibodies which are capable of targetting all FcR, for their ability to mediate target cell destruction. The derivatives are prepared by linking hinge sulphydryl residues via tandem thioether bonds, using a bismaleimide crosslinker: Fab from an anti-FcR mAb linked to Fab from a common anti-target mAb (BsAb), or Fab from the common anti-target mouse antibody linked to human Fc (FabFc or bisFabFc). All the derivatives targetting chick red blood cells gave efficient lysis, although different effector cell donors yielded differences in both the lytic levels achieved and the comparative efficiencies of derivatives. In contrast, significant lysis of the guinea pig lymphoblastic leukaemia, L₂C, regularly resulted only via the anti-FcRIII BsAb and the chimeric derivatives. These results suggest that the chimeric, Fc-containing derivatives mediate tumour cell lysis principally through FcRIII on NK cells. This is in contrast to the situation with the chick red blood cells where the chimeric derivatives appear capable of lysing erythrocytes by utilizing either monocytes or NK cells, because significant (50%) lysis occurred with effector cell populations magnetically depleted through either FcRII or FcRIII. A major difference between these two types of antibody derivative was their ability to function in the presence of high concentrations of normal human Fc. The lysis mediated by BsAb reactive with FcRI or II was unaffected by the presence of human Fc at 2.5 mg/ml (a concentration comparable with that yielded by IgG in plasma) whereas the BsAb recognizing FcRIII and all the Fc-containing derivatives were completely inhibited.This work has been supported by Tenovus, the Cancer Research Campaign, the Leukaemia Research Fund, Italfarmaco, Milano, Italy and the Imperial Cancer Research Fund     

Novel derivatives of 9,10-anthraquinone are selective algicides against the musty-odor cyanobacterium Oscillatoria perornata

Musty "off-flavor" in pond-cultured channel catfish (Ictalurus punctatus) costs the catfish production industry in the United States at least 30 million US dollars annually. The cyanobacterium Oscillatoria perornata (Skuja) is credited with being the major cause of musty off-flavor in farm-raised catfish in Mississippi. The herbicides diuron and copper sulfate, currently used by catfish producers as algicides to help mitigate musty off-flavor problems, have several drawbacks, including broad-spectrum toxicity towards the entire phytoplankton community that can lead to water quality deterioration and subsequent fish death. By use of microtiter plate bioassays, a novel group of compounds derived from the natural compound 9,10-anthraquinone have been found to be much more selectively toxic towards O. perornata than diuron and copper sulfate. In efficacy studies using limnocorrals placed in catfish production ponds, application rates of 0.3 micro M (125 micro g/liter) of the most promising anthraquinone derivative, 2-[methylamino-N-(1'-methylethyl)]-9,10-anthraquinone monophosphate (anthraquinone-59), dramatically reduced the abundance of O. perornata and levels of 2-methylisoborneol, the musty compound produced by O. perornata. The abundance of green algae and diatoms increased dramatically 2 days after application of a 0.3 micro M concentration of anthraquinone-59 to pond water within the limnocorrals. The half-life of anthraquinone-59 in pond water was determined to be 19 h, making it much less persistent than diuron. Anthraquinone-59 appears to be promising for use as a selective algicide in catfish aquaculture.     

DNA-based Fish Species Identification Protocol

We have developed a fast, simple, and accurate DNA-based screening method to identify the fish species present in fresh and processed seafood samples. This versatile method employs PCR amplification of genomic DNA extracted from fish samples, followed by restriction fragment length polymorphism (RFLP) analysis to generate fragment patterns that can be resolved on the Agilent 2100 Bioanalyzer and matched to the correct species using RFLP pattern matching software.The fish identification method uses a simple, reliable, spin column- based protocol to isolate DNA from fish samples. The samples are treated with proteinase K to release the nucleic acids into solution. DNA is then isolated by suspending the sample in binding buffer and loading onto a micro- spin cup containing a silica- based fiber matrix. The nucleic acids in the sample bind to the fiber matrix. The immobilized nucleic acids are washed to remove contaminants, and total DNA is recovered in a final volume of 100 μl. The isolated DNA is ready for PCR amplification with the provided primers that bind to sequences found in all fish genomes. The PCR products are then digested with three different restriction enzymes and resolved on the Agilent 2100 Bioanalyzer. The fragment lengths produced in the digestion reactions can be used to determine the species of fish from which the DNA sample was prepared, using the RFLP pattern matching software containing a database of experimentally- derived RFLP patterns from commercially relevant fish species.Download video file.^{(106M, mp4)}     

The predation impact of juvenile herring Clupea harengusand sprat Sprattus sprattus on estuarine zooplankton

The consumption of estuarine copepods by juvenile herring and sprat during estuarine residency was estimated using fish biomass data and daily rations calculated from two models of feeding in fish: a bioenergetic model and a gastric evacuation model. The bioenergetic model predicted daily rations that were, on average, three times higher than those estimated by a model based on field records of stomach contents. The biomass of herring and sprat in the estuary was negatively correlated with the daily ration suggesting that the clupeid fish populations were resource-limited. Copepod production decreased towards the winter and peaked in spring and summer. The relative importance of predation changed seasonally in function of the migration pattern of herring and sprat. In the spring and the summer, in situ production ofcopepod biomass was higher than the in situ consumption by fish. During the fall and the winter, consumption exceeded production. This suggests that top–down control exerted by marine pelagic fish may be an important force structuring estuarine copepod populations.     

Aggressive and Foraging Behavioral Interactions Among Ruffe

The ruffe, Gymnocephalus cernuus, is a nonindigenous percid in the Great Lakes. Ruffe are aggressive benthivores and forage over soft substrates. Laboratory studies in pools (100cm diameter, 15cm water depth) were conducted to determine whether fish density (low=2, medium=4, high=6 ruffe per pool) changed foraging and aggressive behaviors with a limited food supply of chironomid larvae. All fish densities demonstrated a hierarchy based on aggressive interactions, but ruffe were most aggressive at low and high fish densities. Time spent in foraging was lowest at the low fish density. The best forager at the low fish density was the most aggressive individual, but the second most aggressive fish at the medium and high fish density was the best forager and also the one chased most frequently. A medium fish density offered the best energetic benefits to ruffe by providing the lowest ratio of time spent in aggression to that spent foraging. Based on our results, ruffe should grow best at an intermediate density. With high ruffe densities, we would also expect disparity in size as the more aggressive fish are able to garner a disproportionate amount of the resources. Alternatively, as the Great Lakes are a fairly open system, ruffe could migrate out of one area to colonize another as populations exceed optimal densities.     

Mortality process in relation to aggregation in the southern green stink bug

A simple experiment of simulation was done to analyze the natural mortality process of young larval colonies and egg masses of the southern green stink bug. In this experiment, a degree of contagiousness was allowed in regard to the action of a mortality factor, and was defined as the mean number killed per a colony or an egg mass by the mortality factor within a unit time and the number killed per a colony was assumed to follow the Poisson series with the mean . Thus each component of the Poisson series was opposed to each colony or egg mass which was taken at random from 162 egg masses, 135 and 117 colonies of the first and the second instar larvae, respectively. It was revealed that mortality factors in the field did not act with a small degree of contagiosness, e. g., on all colonies or egg masses, but acted with a large degree of contagiousness, e. g., on some of the colonies or egg masses. Thus differential survival somewhat in all or none way occurred among the insect colonies irrespective of their initial sizes. These results were well explained by taking actual mortality factors into account.     

Strength of coupling between phyto- and zooplankton in Lake Lucerne (Switzerland) during phosphorus abatement subsequent to a weak eutrophication

The strength of coupling between phyto- and zooplankton was measured from 1961 to 1995 by comparing the grazing effect of zooplankton (visible as clear-water phase only in 1968-1994) and also by excluding zooplankton in limnocorral experiments (1980-1984). Although long-term (1961-1995) measurements show little evidence of temporal changes in total biomass of phytoplankton or zooplankton, there is strong evidence of changes in the strength of coupling due to top-down effects. The ratio of change in biomass caused by cladocerans in the intensive grazing period of each year (May/June) and the recovery of netplankton after this period seems to be strongly influenced by the trophic state of the lake. When Lake Lucerne was mesotrophic (1971-1982), the annual mean of monthly changes in phytoplankton biomass was in the range of 1-2, indicating that the biomass more than doubled (or halved) from month to month (no change = 0). Under oligotrophic conditions, the annual average of monthly changes in biomass was below 0.5. Grazing measurements in limnocorrals at 2 m depth with labelled food (Rhodomonas lacustris) showed distinct diel rhythms, with maximum community grazing rate at dusk and dawn. These diel changes were caused by vertical migration of the zooplankton. Grazing rate and zooplankton biomass were strongly coupled, with a maximum rate of 100-200 ml day^-1 mg^-1 (zooplankton biomass) when daphnids were dominant. The decrease in biomass caused by excessive grazing shows parallel trends in nanoplankton and netplankton. However, the increase in biomass after the clear-water phase was largely caused by netplankton.      

Retro-orbital Injection in Adult Zebrafish

Drug treatment of whole animals is an essential tool in any model system for pharmacological and chemical genetic studies. Intravenous (IV) injection is often the most effective and noninvasive form of delivery of an agent of interest. In the zebrafish (Danio rerio), IV injection of drugs has long been a challenge because of the small vessel diameter. This has also proved a significant hurdle for the injection of cells during hematopoeitic stem cell transplantation. Historically, injections into the bloodstream were done directly through the heart. However, this intra-cardiac procedure has a very high mortality rate as the heart is often punctured during injection leaving the fish prone to infection, massive blood loss or fatal organ damage. Drawing on our experience with the mouse, we have developed a new injection procedure in the zebrafish in which the injection site is behind the eye and into the retro-orbital venous sinus. This retro-orbital (RO) injection technique has been successfully employed in both the injection of drugs in the adult fish as well as transplantation of whole kidney marrow cells. RO injection has a much lower mortality rate than traditional intra-cardiac injection. Fish that are injected retro-orbitally tend to bleed less following injection and are at a much lower risk of injury to a major organ like the heart. Further, when performed properly, injected cells and/or drugs quickly enter the bloodstream allowing compounds to exert their effect on the whole fish and kidney cells to easily home to their niche. Thus, this new injection technique minimizes mortality while allowing efficient delivery of material into the bloodstream of adult fish. Here we exemplify this technique by retro-orbital injection of Tg(globin:GFP) cells into adult casper fish as well as injection of a red fluorescent dye (dextran, Texas Red) into adult casper fish. We then visualize successful injections by whole animal fluorescence microscopy.Download video file.^{(55M, mp4)}     

An overview of Acipenseriformes

Acipenseriformes occupy a special place in the history of ideas concerning fish evolution, but in many respects, phylogenetic studies of the group remain in their infancy. Even such basic questions as the monophyly of Acipenser (the largest genus) are unanswered. We define relationships based on comparative osteology, which allows us to incorporate well-preserved fossils into analyses. Acipenseriformes has existed at least since the Lower Jurassic (approximately 200 MYBP), and all fossil and recent taxa are from the Holarctic. Phylogenetic relationships among Paleozoic and Early Mesozoic actinopterygians are problematic, but most workers agree that Acipenseriformes is monophyletic and derived from some component of paleonisciform fishes. (Paleonisciformes is a grade of primitive non-neopterygian actinopterygians, sensu Gardiner 1993.) Taxa discussed in comparison here are: Cheirolepis, Polypterus, Mimia, Moythomasia, Birgeria, Saurichthys, Lepisosteus and Amia. We review generic diversity within the four nominal families of fossil and recent Acipenseriformes (Chondrosteidae, Peipiaosteidae, Polyodontidae, and Acipenseridae), and provide a cladogram summarizing osteological characters for those four groups. Monophyly of the two extant families is well-supported, but there are no comprehensive studies of all of the known species and specimens of Chondrosteidae and Peipiaosteidae. As a result, sister-group relationships among Chondrosteidae, Peipiaosteidae, and Acipenseroidei (= Polyodontidae + Acipenseridae) are unresolved. We discuss five features fundamental to the biology of acipenseriforms that benefit from the availability of our new phylogenetic hypothesis: (1) specializations of jaws and operculum relevant to jaw protrusion, feeding, and ram ventilation; (2) anadromy or potamodromy and demersal spawning; (3) paedomorphosis and evolution of the group; (4) the bioégeography of Asian and North American polyodontids and scaphirhynchines; and (5) the great abundance of electroreceptive organs in the rostral and opercular regions. Finally, we summarize our nomenclatural recommendations.     

Locomotor response of the goldfish to polarized light and itse-vector

Summary The locomotor orientation of eleven goldfish, 20–25 cm long, was monitored during periods varying between 24 hours and 8 1/2 days, to verify the response to a depolarized and polarized sky, 100 cm in diameter, and to abrupt 90 ° degree rotation of thee-vector. The monitor consisted of a cylindrical tank with 16 peripheral compartments (Fig. 1) to which the fish had free access. Entry into and exit from each compartment was electronically recorded. The distribution of entries, which had no cyclical relationship with the compartments in depolarized light, became significantly symmetrical and bimodal in polarized light with the preferred compartments oriented parallel with thee-vector. Abrupt 90 ° rotation of the vector counter clockwise maintained this relationship during the entire duration of the recordings (up to 17 hours) (Fig. 2). The mean of the orientation angles of the fish on leaving compartments aligned with thee-vector were significantly higher than those from the remaining compartments (Fig. 3). This behavior tended to keep the locomotor orientation parallel with thee-vector as the fish moved between compartments. A strong cyclical relationship between these orientation angles and the compartments of origin was present in polarized but absent in depolarized light. Counter clockwise 90 ° rotation of thee-vector maintained the cyclic behavior of angles but the relationship between the larger means and thee-vector shifted over one or two compartments. This shift disappeared in clockwise rotation. This phenomenon may be due to one of these directions being unnatural. The results demonstrate a pronounced sensitivity and response toe-vector orientation in the goldfish. The sensory mechanism remains unknown.The authors are greatly indebted to Dr. T. H. Waterman for a critical review and discussion of the results here presented.     

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the visual scene. However, the OKR requires that an animal detect moving stripes and it is possible that fish that fail to exhibit an OKR may not be completely blind. One zebrafish mutant, the no optokinetic response c (nrc) has no OKR under any light conditions tested and was reported to be completely blind. Previously, we have shown that OFF-ganglion cell activity can be recorded in these mutants. To determine whether mutant fish with no OKR such as the nrc mutant can detect simple light increments and decrements we developed the visual motor behavioral assay (VMR). In this assay, single zebrafish larvae are placed in each well of a 96-well plate allowing the simultaneous monitoring of larvae using an automated video-tracking system. The locomotor responses of each larva to 30 minutes light ON and 30 minutes light OFF were recorded and quantified. WT fish have a brief spike of motor activity upon lights ON, known as the startle response, followed by return to lower-than baseline activity, called a freeze. WT fish also sharply increase their locomotor activity immediately following lights OFF and only gradually (over several minutes) return to baseline locomotor activity. The nrc mutants respond similarly to light OFF as WT fish, but exhibit a slight reduction in their average activity as compared to WT fish. Motor activity in response to light ON in nrc mutants is delayed and sluggish. There is a slow rise time of the nrc mutant response to light ON as compared to WT light ON response. The results indicate that nrc fish are not completely blind. Because teleosts can detect light through non-retinal tissues, we confirmed that the immediate behavioral responses to light-intensity changes require intact eyes by using the chokh (chk) mutants, which completely lack eyes from the earliest stages of development. In our VMR assay, the chk mutants exhibit no startle response to either light ON or OFF, showing that the lateral eyes mediate this behavior. The VMR assay described here complements the well-established OKR assay, which does not test the ability of zebrafish larvae to respond to changes in light intensities. Additionally, the automation of the VMR assay lends itself to high-throughput screening for defects in light-intensity driven visual responses.Download video file.^{(107M, mp4)}     

Risk from genetically engineered and modified marine fish

In support of the emerging industries of warmwater marine fish mariculture, genetic engineering and classical genetic improvement programmes have been initiated for a variety of exclusively marine fish. These programmes have the potential to perturb allele and genotype frequencies, or introduce novel alleles and genes into conspecific wild populations. Despite concerns to the contrary, the following hypothesis remains to be falsified: laboratory induced allele frequency/genotype changes and novel alleles or genes have a negligible probability of being selectively favoured in wild populations under natural selection, and accordingly, without sustained large scale releases, have little potential for ecological impact     

Seawater Sampling and Collection

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses. For today''s demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. An A-frame derrick, with a multi-purpose winch and cable system, is used in combination with Niskin or Go-Flo water sampling bottles. Conductivity, Temperature, and Depth (CTD) sensors are also used to sample the underlying water mass. To minimize outgassing, trace gas samples are collected first. Then, nutrients, water chemistry, and cell counts are determined. Finally, waters are collected for biomass filtration. The set-up and collection time for a single cast is ~1.5 hours at a maximum depth of 215 meters. Therefore, a total of 6 hours is generally needed to complete the collection series described here.Download video file.^{(98M, mp4)}