Rhythmic settling induced by temperature cycles in continuously-stirred autotrophic cultures of Euglena gracilis (Z strain)

Summary Temperature cycles which produced synchrony of cell division in autotrophic Euglena (preceding paper, Terry and Edmunds, 1970) usually also evoked a cellular settling rhythm. The rhythm was expressed as a recurrent cycle of cell attachment to culture-vessel surfaces, with nearly the same phase angle to each of the different temperature cycles. Attachment occurred in cultures stirred rapidly enough for thorough mixing of the cell suspension, but it was possible to prevent attachment by increased agitation. The settling rhythm was entrained by temperature cycles with periods shorter than the minimum period length required to phase cell division, but the rhythm also persisted with a circadian period at constant temperature in continuous light following entrainment by 12,12 hr temperature cycles. The rhythm appeared in stationary-phase cultures exposed to either cold-synchronizing or heat-synchronizing temperature cycles, and also in growth-phase cultures exposed to cold-synchronizing cycles. The settling rhythm was generally not observable in heat-synchronized growth-phase cultures although it often appeared in the same cultures as they approached the stationary phase.This work is derived from a dissertation submitted to the Department of Biological Sciences of the State University of New York at Stony Brook by OzTerry in partial fulfillment of the requirements for the degree of Doctor of Philosophy. It was supported in part by a National Science Foundation Cooperative Graduate Fellowship to 0. Terry and in part by National Science Foundation research grants GB-4140 and GB-6892 and by State University of New York Research Foundation Grant-in-Aid 31-7150A to L. Edmunds.     

The coupling effects of some thiol and other sulfur-containing compounds on the circadian rhythm of cell division in photosynthetic mutants of Euglena

Previous work has demonstrated a persisting, free-running, circadian rhythm of cell division in the P₄ZUL photosynthetic mutant of the alga Euglena gracilis Klebs (Strain Z) Pringsheim grown organotrophically in continuous light or darkness at 19° C following prior synchronization by a repetitive LD: 10,14 light cycle. A similar circadian rhythmicity has been recently discovered in the W₆ZHL heat-bleached and the Y₉ZNalL naladixic acid-induced mutants of Euglena grown under comparable conditions. Over extended timespans, however, these mutants appear to gradually lose first their ability to display persisting overt rhythms, and then even their capability of being entrained by imposed LD cycles. These properties can be restored by the addition of certain sulfur-containing compounds to the medium including cysteine, methionine, dithiothreital, sodium monosulfide, sodium sulfite, and sodium thiosulfate, as well as thioglycolic [mercaptoacetic] acid. The implications of these findings toward biological clock mechanisms are discussed: It appears that some sort of coupling process is operating as opposed to the initiation of an underlying oscillation.Non-Standard Abbreviations LL continuous illumination - DD continuous darkness - LD repetitive light-dark cycle - SS stepsize - period of biological rhythm Supported by research grants (GB-36287, GB-43543) from the National Science Foundation.Reports on portions of this work were presented at the 19th Annual Meeting of the Biophysical Society, Philadelphia, Pennsylvania, February 19–21, 1975; at the XII International Botanical Congress, Leningrad, U.S.S.R., July 3–10, 1975; and at the XII International Conference of the International Society for Chronobiology, Washington, D.C., August 10–13, 1975.     

Ambient temperature cycles entrain the free-running circadian rhythms of the stripe-faced dunnart,Sminthopsis macroura

Summary We examined the effect of cycles of 12 h warm (35 ± 2 °C) and 12 h (21 ± 2 °C) ambient temperature (Ta) upon the circadian activity rhythms of stripe-faced dunnarts, Sminthopsis macroura, free-running in conditions of constant dark (DD) or constant light (LL). It was hypothesized that dunnarts would entrain to the temperature cycles (TaHLs) or show perturbations of period, and that LL would act synergistically with the TaHLs in these effects. Under DD, 2 of 6 animals showed clear entrainment to the TaHLs. Other animals exhibited changes of period () and heavy negative masking of activity during the warm fraction of the TaHLs. Under LL, 3 of 12 animals entrained to the TaHLs. It was concluded that Ta is a significant though weak Zeitgeber for S. macroura compared to light. It is possible that TaHLs entrain homeotherm activity rhythms by altering the rhythm of body temperature, which is usually tightly coupled to activity.Abbreviations TaHL a cycle of Higher and Lower ambient temperature - TaC Constant Ta - Tb body temperature     

Entrainment and phase shifting of the circadian rhythm of cell division by light in cultures of the achlorophyllous ZC mutant ofEuglena gracilis

The photosynthesis-deficient ZC mutant ofEuglena gracilis Klebs (strain Z) was cultured at 16°C on an aerated, magnetically stirred, mineral medium containing 0.1% ethanol (pH 7.0). Cell division could be entrained by a 12: 12 light: dark cycle (LD: 12, 12) or even by a one-pulse skeleton photoperiod (LD: 1,23) The rhythm free-ran in DD for at least 8 days with a circadian period (=25.5 h) in populations that had been previously entrained by LD. The freerunning rhythm could be phase-shifted by a single 1-h light pulse (3000 lx). The strong (Type 0) phase-response curve derived from the resetting effects of such signals given at different circadian times was similar to that for the photosynthetic wild-type strain. These results demonstrate that the presence of a functional chloroplast compartment is not necessary for the circadian clock to function inEuglena and suggest that phase resetting of the circadian clock by light occurs via a similar pathway in both photosynthetic and non-photosynthetic cell types.     

The effect of photoperiod on daily rhythms of Oxygen consumption in the tropical toad,Bufo marinus

Summary Oxygen consumption was measured with an automatic continuously recording electrolysis system in toads acclimated at 15° C and under photoperiods of LD 816, 1212, 168, 66, 231 and LL and DD. All groups exposed to LD had pronounced significant daily rhythms with a trough near the onset of the photophase and a peak at the beginning of the scotophase (Figs. 1–3, 9, 10), while no rhythms were detected in animals exposed under free-running conditions of constant light (Figs. 6–7) or constant darkness (Fig. 8), even on the first day under LL or DD. These rhythms are thus shown to be non-circadian, since they do not persist in freerunning conditions of up to 45 days. Some preliminary evidence from studies on locomotor activity (Figs. 12, 13) indicates that the daily rhythm in metabolic rate may be independent of the locomotor activity cycle. The difference in the rate of oxygen consumption during peak and low hours of each daily cycle is defined as the daily routine metabolic scope, which may be more useful in ecological studies of animal energetics than the difference between the minimum and a forced maximum metabolic rate. No correlations were found between daily changes in atmospheric pressure and cycles of oxygen consumption.This work was supported by a grant from the National Science Foundation (GB-3574) and a University of Rhode Island Research Committee Grant-in-aid. We are grateful to Robert Cubert for aid in designing and constructing the electrolysis system and for assistance with computer programs.     

Phosphopyruvate carboxylase activity and carbon dioxide fixation via C4 acids over the division cycle in synchronized Euglena cultures

Summary Phosphoryruvate carboxylase activity was determined in division synchronized Euglena gracilis strain Z cultures. The profile of enzyme activity was essentially that of a peak enzyme; activity increased over the light phase of the cycle, doubling by early dark phase followed by a substantial decline in activity near the end of the dark phase. Dark carbon dioxide fixation did not parallel changes in phosphoryruvate carboxylase activity. The rate of carbon dioxide fixation increased fourfold over the light phase but decreased in the dark phase until it was only double the rate at the beginning of the light phase.Although the specific activity of phosphopyruvate carboxylase was greater than that of ribulose 1–5 diphosphate carboxylase in Euglena cell extracts at all stages over the division cycle C₄ acids were not an early product of carbon dioxide fixation in the light, neither did they ever account for more than a small proportion of the total ¹⁴C present in the soluble fraction of the cells. Phosphopyruvate carboxylase was shown by the non-aqueous localization technique to be present in the cytoplasm in Euglena, and it is concluded that the main function of this enzyme in algal cells is to provide an anaplerotic sequence to the tricarboxylic acid cycle.     

Wall structure of a bitunicate ascus

Summary Ultraviolet light-induced, bleached Euglena clones exhibit synchronous steps of cell division in response to daily cycles of light and dark. The cyclic division activity, in the bleached cells, will persist in constant lighting conditions with a period, independent of temperature, of about 24 h. This persisting rhythm of cell division supports the hypothesis that this phase of the cell cycle may be coupled to the fluctuations of the endogenous circadian clock of the cell.Newly isolated bleached clones are sensitive to light in their growth rates and metabolic characteristics, showing light induced difference in substrate-stimulated respiration, and production of the polyglucan, paramylon. After repeated subculturing of a bleached clone the photosensitivity of the metabolic characteristics and of the growth rate are diminished along with the ability to photo-entrain division synchrony. Division control and the induction of cell synchrony in this organism apparently involve both the temporal influence of the endogenous cell clock and one or more other photosensitive reactions in the metabolism of the cell.A unique culture mixing technique utilizing the bleached Euglena, failed to support the hypothesis of the involvement of intercellular communication in the maintenance of cell synchrony in constant lighting conditions.     

Dynamics of pattern formation

Experiments were designed to test the validity ofTuring's suggested pattern-forming mechanisms, which are initially capable of giving rise to only five to seven uniform structures.TheOregon-R (wild-type), mass-cultured strain ofDrosophila melanogaster was employed. Selection for the regular arrangement of microchaetac on the margin of the fourth abdominal sternite was practiced for twenty generations. In the L line, individuals with six uniformly spaced bristles were selected as parents of every generation. Due to the absence of nine bristles dividing the sternal margin uniformly, the progeny was raised in each generation in the H line from males and females with nine as equidistant bristles as possible. The whole experiment was performed at 25±0.50°C.Selection was effective in increasing the frequency of six regular bristles in the L line. But no progress in the desired direction was obtained in the H line, although the proportion of sternites with nine irregular structures was found to increase. The experimental results supportTuring's diffusion-reaction scheme of pattern formation in morphogenesis.Supported by grants GB-1388 and GB-3219 from the National Science Foundation.     

The S phase is discrete and is controlled by the circadian clock in the marine dinoflagellate Gonyaulax polyedra

Cloned cultures of the dinoflagellate Gonyaulax polyedra grown in a 12-h light-12-h dark cycle (LD 12:12) were synchronized to the beginning of G₁ by a two sequential filtration technique. After the second filtration, with the cultures growing in LD 12:12, not many cells had divided after 1 day, but approximately half underwent cell division after 2 days. Flow cytometric measurements of the cells revealed that there is one unique S phase starting about 12 h prior to cell division and lasting for less than 4 h. A majority of cells in cultures synchronized in the same way but maintained in continuous light (LL) after filtration also divided synchronously after 2 days. Just as for the cultures in LD 12:12, those in LL have a similar discrete DNA synthesis phase prior to division. It is concluded that the circadian control of cell division acts before the S phase, giving rise to a discontinuous DNA synthesis phased by the circadian clock.     

A covert contaminant of cultured plant cells: elimination of a Hyphomicrobium sp. from cultures of Datura innoxia (Mill.)

We have discovered a bacterial contaminant in some cell cultures of Datura innoxia (Mill.). The bacterium was tentatively identified as a species of Hyphomicrobium on the basis of its morphology and life cycle, and was isolated and grown in pure culture on a defined medium. The contaminant was not macroscopically observable in plant cell cultures. It caused neither a reduction of plant cell growth nor a noticeable increase in culture turbidity. Furthermore, it was not readily detectable by many standard assays for culture contamination: it would not grow alone in plant culture medium or yeast extract potato dextrose medium, and grew only very slowly on nutrient agar or beef-peptone medium. Repeated treatments with a combination of streptomycin (100 g/ml) and carbenicillin (100 g/ml) eliminated the contaminant from D. innoxia cell cultures without harming the plant cells.     

Spatial distribution of 0-group eel larvae (Anguilla sp.) in the Sargasso Sea

Both Atlantic eel species (Anguilla anguilla andA. rostrata) were collected in the Sargasso Sea during the 1979 cruise of the F. R. V. Anton Dohrn and R. V. Friedrich Heincke. A total of 3,097 0-group larvae were caught during 80 hauls using the Isaacs Kidd Midwater Trawl (55 hauls) and the 9-fold opening and closing net MOCNESS (25 hauls). 11 hauls of the MOCNESS indicated that the larvae showed a preference for the 150–175 m water depth during daytime and for the 50–75 m depth during night. The northern distribution limit seems to coincide with the 18°C-water at the sea surface. The Antilla current characterised by higher temperatures and salinities could have been the southern distribution limit. The north-easternmost occurrence ofA. anguilla was noted at 50°W; the eastern distribution limit of this species could be farther east. The westernmost station at 69°W was positive for both species.A. anguilla, therefore, is very likely to occur beyond this area. The easternmost occurrence ofA. rostrata was noted at 52°W though sporadic and with increasing abundance towards the west. At the western end of the network of stations the highest concentration of larvae from both species was recorded. Oceanographic investigations reveal that the distribution of the smaller larvae (size-groups <7 mm and <10 mm) almost coincides with the assumed spawning area.     

Energy conservation in whole cells of the methylotrophic bacterium Methylophilus methylotrophus

Respiratory chain phosphorylation has been investigated in the methylotrophic bacterium Methylophilus methylotrophus following the addition of oxidisable substrates to aerobic, whole cell suspensions. Initial-rate experiments showed that ATP synthesis occurred at the overall expense of AMP and inorganic phosphate via the sequential action of the ATP phosphohydrolase and adenylate kinase; some of the nascent ATP was rapidly used to synthesis nonadenine nucleoside triphosphates. After being corrected for ATP turnover, Pi/O quotients of 0.46 to 0.54, 0.77 and 1.37 nmol/ng-atom O were obtained for the oxidation of methanol dehydrogenase-linked substrates (methanol, ethanol and acetaldehyde), duroquinol and formate (NAD⁺-linked) respectively. These values were proportional to the H⁺/O and/or K⁺/O quotients exhibited by these substrates, and yielded an average H⁺/ATP (H⁺/Pi) quotient of 4.2 ng-ion H⁺/nmol. Steady-state experiments showed that the extent of cellular energisation varied with the respiration rate but was always in the order methanol > duroquinol > acetaldehyde, thus indicating that under these longer-term conditions methanol was completely oxidised to yield PQQH₂ and 2NAD(P)H. These results are discussed in terms of the various reactions which lead to the generation or utilisation of the protonmotive force in this organism.Abbreviations FCCP carbonylcyanide p-trifluoromethyxyphenyl-hydrazone - bulk phase, transmembrane electrochemical potential difference of protons ( ) - pH bulk phase, transmembrane pH difference (pH_in–pH_out) - bulk phase, transmembrane electrical potential difference (_in - _out) - [P] concentration of anhydride phosphate bonds in adenine nucleotides (2[ATP]+[ADP]) - FPLC fast protein liquid chromatography - PQQ pyrroloquinoline quinone - Gp phosphorylation potential     

Synchronous cell divisions in Anacystis nidulans Richter

Summary The blue-green alga, Anacystis nidulans was successfully synchronized in continuous light (15,000 lux) by a 14 hr cycle, consisting of 8 hrs at 26°C, and 6 hrs at 32°C, coupled with periodic dilution of the algal suspension to a constant cell number at the end of each cycle. The alga continues to grow at the lower temperature, and a division burst begins 2 hrs after transfer to the higher temperature.

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Zusammenfassung Die Cyanophycee Anacystis nidulans wurde in Dauerlicht von 15 000 Lux vollständig synchronisiert. Die Cyclen bestehen aus 8 Std bei 26° C und 6 Std bei 32° C, an ihrem Ende wurde jeweils mit frischer Nährlösung auf die Ausgangszellzahl von 3·10⁷ Zellen/ml verdünnt. Bei beiden Temperaturen kann sich die Alge vermehren, unter den synchronisierenden Bedingungen des Temperaturwechsels beginnt der Teilungsschub (Verdopplung der Zellzahl) bei 32° C nach 2 Std.

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Pringsheim (1968) renamed this alga as Lauterbornia nidulans.     

Effect of light and temperature on the bleaching of the yellow mutant y-1 ofEuglena gracilis

Summary The yellow mutant y-1 ofEuglena gracilis, whose plastids are chlorophyll-deficient, is stable only in the dark and at suboptimal growth temperatures (below 26 C).At growth temperature of 30 C (in darkness) this strain is unstable and undergoes permanent bleaching. The process is enhanced by light so that bleaching may be increased up to 100 per cent. The degree of bleaching is proportional to light intensity.Bleaching is possible only during the active cell division. In the resting medium there is no bleaching, but total carotenoid synthesis is stimulated by illumination. At suboptimal growth temperature (26 C) the mutant is bleached only in the light, if light intensity exceeds 2,000 Lux. The results indicate that the process of bleaching in the yellow mutant is much more sensitive to temperature and light than in wild typeEuglena.     

Life cycles of marine nematodes

Summary Monhystera denticulata Timm, a free-living nematode present in the aufwuchs assemblages of several marine macrophytes located in North Sea Harbor, Southampton, New York, was isolated from Zostera marina and established in laboratory culture in order to study the influences of temperature and salinity on its life history. Under experimental conditions, M. denticulata has a generation time (Measured as the time elapsing between the first egg depositions of consecutive generations) of 10–12 days at 25° C and 26 S, which represent optimal growth conditions in the laboratory. The organism has a generation time of 20 days at 25° C and 13, 17 days at 25° C and 39, 18 days at 15° C and 26, 36 days at 15° C and 13 and 34 days at 15° C and 39. As conditions vary from the optimum of 25° C and 26 S, a decrease in temperature of 10° C and an increase or decrease in salinity of 13 results in a doubling of the generation time. At 5° C the generation time is about 180–197 days.Assuming optimum conditions and average generation time, about 15 generations of M. denticulata could occur in North Sea Harbor during the year. The number of generations occurring in reality is probably less, however, due to the fact that the females deposit their eggs over a period of several days.This work was supported by National Science Foundation Grant GB-19245.Contribution No. 04 from the Institute of Oceanography, City University of New York.     

Circadian rhythms of corneal mitotic rate,retinal melatonin and immunoreactive visual pigments,and the effects of melatonin on the rhythms in the Japanese quail

We investigated circadian ocular rhythms in the Japanese quail, Coturnix coturnix japonica. The birds were placed under light-dark cycles (LD 1212), constant light (LL) and constant darkness (DD), and the retinas were dissected out at four-hour intervals throughout 24 h. Following measurements were performed. (1) Melatonin content in the retina was measured by radioimmunoassay. It was low in light and several folds higher in darkness under LD 1212. The rhythm continued in DD, but disappeared in LL. (2) Mitotic figures in the corneal epithelium were counted. Similar rhythms to the melatonin content were observed in the corneal mitotic rate with a slight phase delay. (3) The retinas were fixed at 4-h intervals and immunostained with anti-bovine rhodopsin serum and anti-chicken iodopsin monoclonal antibodies. The outer segments of photoreceptor cells were stained intensively throughout 24 h in LD 1212, LL and DD. In contrast, the stainability of the locus close to the outer limiting membrane where the Golgi apparatus exists changed diurnally. Scores showing the ratio of cells with positive staining indicated high values from 4 h after the onset of light to the beginning of dark phase under LD 1212. The values were high throughout 24 h in LL and intermediate or low in DD. (4) To investigate the effect of melatonin on the corneal mitotic rate and visual pigments at the Golgi region, melatonin was injected into one eye and saline into the contralateral eye. Melatonin induced a phase advance in the corneal mitotic rate under LD 1212, but did not induce a rhythm under LL. The ratio of photoreceptor cells with positive staining to anti-visual pigment antibodies at the Golgi region was not affected by melatonin injection.Abbreviations ANOVA analysis of variance - BSA bovine serum albumin - DD constant darkness - Io-mAb monoclonal antibodies against chicken iodospin - LD light-dark - LL constant light - mRNA messenger ribonucleic acid - PBS phosphate buffer solution - Rh-As antiserum against bovine rhodopsin - SCN suprachiasmatic nucleus - T transducin - T transducin      

Pineal melatonin rhythms in the lizardAnolis carolinensis: effects of light and temperature cycles

Summary Pineal and ocular melatonin was assessed, over 24 h periods, in male lizards (Anolis carolinensis) entrained to 24 h light-dark (LD) cycles and a constant 32 C, and in lizards entrained to both 24 h LD cycles and 24 h temperature cycles (32 C/20 C). At a constant temperature, the duration of the photoperiod has a profound effect on the duration, amplitude, and phase of the pineal melatonin rhythm (Fig. 1). The pineal melatonin rhythm under cyclic temperature peaks during the cool (20 C) phase of the cycle regardless of whether or not the cool phase occurs during the light or dark phase of a LD 1212 cycle (Fig. 3). Under a temperature cycle and constant dim illumination, a pineal melatonin rhythm is observed which peaks during the cool phase of the temperature cycle, but the amplitude of the rhythm is depressed relative to that observed under LD (Fig. 2). Illumination up to 2 h in duration does not suppress the nocturnal melatonin peak in theAnolis pineal (Fig. 4). No melatonin rhythm was observed in the eyes ofAnolis under either 24 h LD cycles and a constant temperature (Fig. 1), or under simultaneous light and temperature cycles (Fig. 3). Ocular melatonin content was, in all cases, either very low or non-detectable.Abbreviations HIOMT hydroxyindole-O-methyltransferase - NAT N-acetyltransferase     

Response characteristics of cold cell on the antenna ofLocusta migratoria L.

Summary The dendritic outer segment of the cell which is most likely the cold unit in the poreless coeloconic sensilla onLocusta migratoria antennae, has finger-like projections up to 1.5 m long and 0.13 m thick (Fig. 1). This unit responds to constant temperature, to slowly changing temperature and to step changes. Under stationary conditions impulse frequency attained 35 imp/s. Between 14 °C and 41 °C the higher frequencies were associated with the higher temperatures (Fig. 5). In this range the differential sensitivity is positive but not large: + 0.8 (imp/s)/°C. Its resolving power for steady temperature is 4.7 °C.Downward step changes produced by shifting between airstreams at different temperatures yield far higher frequencies (Figs. 2, 3). Step amplitudes were between –0.1 °C and –12 °C; the conditioning temperature from which the steps were initiated, was between 16 °C and 33 °C. Frequency peaked during the first 50 ms after stimulus onset (Fig. 2) and reached its highest values (310–340 imp/s) at initial temperatures above 30 °C and steps larger than –10 °C (Fig. 4). The mean differential sensitivity from 23 curves was –19 (imp/s)/°C and the resolving power 0.6 °C.During slowly changing temperature the impulse frequency was governed by two parameters simultaneously: ambient temperature and its rate of change. Rates were between 0.001 °C/s or less, and 0.03 °C/s in either direction. Frequency was higher during slow cooling at a given temperature than during slow warming (Fig. 6). The average differential sensitivity to the rate of change was –210 (imp/s)/(°C/s). Further, the larger responses to cooling developed at lower ambient temperatures (differential sensitivity: –1.0 (imp/s)/°C). It is to be noted that this sign is negative, in contrast to the sign for differential sensitivity to constant temperature and also for the influence of initial temperature on the response to downward step changes.Abbreviations b Slope of characteristic curve, differential sensitivity - F impulse frequency in imp/s - imp/s impulses/s - P _w partial pressure of water vapor in torr - r correlation coefficient - T temperature in °C - T T-step - x resolving power in °C     

Effects of “skeleton” photoperiods and high frequency light-dark cycles on the rhythm of cell division in synchronized cultures of Euglena

Summary Two further lines of evidence support the contention (Edmunds, 1966) that the cell cycle in autotrophically grown Euglena can be coupled to an endogenous, circadian biological clock under certain conditions. So-called skeleton photoperiods (LD: 3,6,3:12 and LD: 4,4,4:12) following a complete photoperiod regime entrain the cell division rhythm in the population to a precise 24 hr period, although the step-sizes of the successive fission bursts are always less than 2.00, indicating that not all cells divide in any one 24 hr interval. These findings imply that the continuous action of light is not required for synchronization and suggest that the putative oscillation underlying the rhythm can be phased by discrete light (or dark) pulses or signals.The effects of high frequency LD cycles whose periods were integral submultiples of 24 hr were also investigated. In most regimes (LD:1/4,1/2; LD:1/2,1; LD: 1,2; LD: 1,3; LD: 2,4; LD: 2,6; LD: 4,4) synchronous cell division iccurred in the culture with an average period of 26–27 hr, although only a fraction of the cells divided during any one burst. Similar results were obtained if (i) a synchronized culture was exposed to certain high frequency cycles whose periods were not integral submultiples of 24 hr (e.g., LD: 5,5 or LD: 8,8); (ii) an asynchronous culture (grown in LL) was subsequently exposed to a high frequency cycle; or (iii) a synchronized culture was subjected to a random LD cycle. The synchrony does not break down as long as the given LD regime is imposed and shows some indications of persistence in certain ensuing conditions of continuous illumination.A general formula was derived which predicts the time of division, t _D , for an individual cell: t _D =k+n, where k is the initial phase delay, n is an integer, and is the free-running period of the rhythm observed in the population. These results are interpreted as indicating that the high frequency cycles employed were unable to entrain the circadian oscillation(s) hypothesized to underly and gate cell division, with the result that the rhythm reverted to its free-running period. Exposure to such cycles, however, apparently either initiates a rhythm or synchronizes the phases of the individual oscillations in the populations of cells. The possible direct interaction between energy supply and the observed somewhat variable period lengths is discussed; also, the relevance of stochastic models for the decay of division synchrony in the absence of a recurrent synchronizing procedure is considered.Some of these results were initially reported at the 5th International Congress on Photobiology, Hanover, N.H., U.S.A., August 26–31, 1968.This work was supported by NSF research grants #GB-4140 and #GB-6892 to L. Edmunds.     

A new species of Gastrosaccus (Crustacea,Mysidacea) from Algoa Bay (South Africa)

The effect of temperature on growth rate, shell size and shell shape in Krithe praetexta praetexta (Sars) was studied in four thermocultures. From July 1995 to June 1996, the cultures were kept in a continuously flowing open system pumping water from the intermediate watermass of the Gullmarn fjord, west coast of Sweden. Three cultures were kept at constant temperatures of 5, 10 and 14 °C, respectively. The fourth (reference) culture largely followed the natural variation in temperature. At the termination of the experiment, all living ostracods from a 125 m sieve were sampled from the cultures. Population age structures were analysed for the various thermocultures of K. praetexta praetexta. These were more shifted towards later ontogenetic stages with higher temperature, i.e. the ontogenetic development was more rapid in the warmer cultures. An alternative explanation is due to diapause causing cohorts to accumulate in some ontogenetic stages only when the temperature is constant. The differences in shell size of K. praetexta praetexta among the thermoconstant cultures were not statistically significant.