GRAZER-INDUCED COENOBIAL FORMATION IN CLONAL CULTURES OF SCENENDESMUS OBLIQUUS (CHLOROCOCCALES, CHLOROPHYCEAE)

Five strains of the phenotypically plastic alga Scenedesmus obliquus (Turpin) Kützing were cultured in standard medium or in medium containing filtered water from a Daphnia culture to examine grazer-induced coenobia formation. Strains varied considerably in their response to Daphnia chemicals. A rapid formation of four- and eight-celled coenobia was observed in the presence of Daphnia water in three strains. One strain showed numerous irregular aggregates with more than eight cells per colony, and no colony formation occurred in a fifth strain. In standard medium, all cultures remained unicellular. Growth rates were affected by Daphnia water in four strains and were different among strains. The chl a concentration was significantly different among strains but was not influenced by Daphnia water. The specific absorption coefficient was reduced at larger colony sizes, suggesting a possible cost associated with colony formation. The phenomenon of grazer-induced coenobia formation seems widespread but not universal within the species S. obliquus.     

EFFECT OF GRAZING-ASSOCIATED INFOCHEMICALS ON GROWTH AND MORPHOLOGICAL DEVELOPMENT IN SCENEDESMUS ACUTUS (CHLOROPHYCEAE)

Populations of the phenotypically plastic alga Scenedesmus acutus Meyen were cultured in standard medium or in medium with filtered water from a Daphnia culture to examine the expression of ecomorphs in both water types. A rapid formation of four- and eight-celled coenobia was observed in the presence of Daphnia water, but not in standard medium. Moreover, cell dimensions were increased in the Daphnia water containing medium. Population growth rates were comparable in both water types; however, the carrying capacity appeared significantly lower in Daphnia water. Daphnia needed to feed on digestible food to produce the colony-inducing chemicals; medium from starved animals and Daphnia fed polystyrene beads appeared inactive. Neither suspensions of homogenate of Scenedesmus and Cryptomonas nor auxins affected colony formation or growth rate in S. acutus. The colony-inducing infochemicals are probably not constituents of the algae themselves; however, modification of algal constituents in the digestive system of daphniids may be involved.     

Does microcystin disrupt the induced effect of Daphnia kairomone on colony formation in Scenedesmus?

In natural aquatic system, Scenedesmus and Microcystis species usually coexist. Microcystins are released into water after lysis of Microcystis cells during the collapse of heavy blooms. The released toxins can then come into contact with a wide range of aquatic organisms. In this study, we used filtered Daphnia test water containing kairomone from Daphnia magna to stimulate the inducible colony formation in Scenedesmus obliquus under microcystin-contaminated system, to examine how microcystin affects the induced effect of Daphnia kairomone on colony formation in S. obliquus. The results showed neither microcystin nor Daphnia kairomone affected the growth of S. obliquus. Microcystin neither promoted nor impaired the overall Daphnia-induced colony formation in S. obliquus, except reducing the proportion of eight-celled colonies on day 2, indicating that the effect of microcystin was just short-term and in general did not disrupt grazer-induced colony formation of S. obliquus.     

The effect of substances from different zooplankton species and fish on the induction of defensive morphology in the green alga Scenedesmus obliquus

The common green alga Scenedesmus obliquus may respond morphologicallyto the presence of natural enemies. Exposure to water-bornecues from the herbivorous zooplankton Ceriodaphnia reticulata,Daphnia galeata x hyalina, Daphnia magna and Daphnia pulicariastimulated the formation of protective colonies in S. obliquus.This response seemed strongly related to the amount of algaegrazed upon, because a highly significant correlation betweenthe amount of S. obliquus consumed and the induced colony formationwas found. However, when exposed to medium that had been inhabitedby the carnivorous zooplankton Bythotrephes longimanus and Leptodorakindtii, no colony formation occurred. A similar result wasobtained in two different experiments when S. obliquus was exposedto filtrate from cultures of the freshwater fish ide (Leuciscusidus) and perch (Perca fluviatilis). These results support thehypothesis that S. obliquus responds to a herbivorous zooplanktonchemical cue, rather than to a more general animal excretoryproduct. No support was obtained for the hypothesis that cuesfrom the enemy of their enemy could serve as signals to thealgae: despite the presence of filtrate from fish culture, filtratefrom Daphnia cultures induced the formation of colonies in S.obliquus. The biological activity seems to be linked to thealga–grazer interaction, which ensures a reliable cuethat evokes the morphological response of S. obliquus only whennecessary. The grazer-induced colony formation can be viewedas an adaptive reaction in habitats with variable grazing pressurefrom an assemblage of many different herbivores to pare downmortality through grazing.     

Psychrotrophic Clostridia

Of 287 strains of clostridia of 23 different species 96 grew well at 6°, 77 of which were Clostridium carnofoetidum var. amylolyticum and 16 were Cl. perfringens.
The water of polluted wells seems to be the normal habitat of Cl. carnofoetidum var. amylolyticum . This species is both proteolytic and saccharolytic and is not pathogenic for mice or guinea pigs. Spores survive at 80° for 40 min. Mg is essential to obtain sporulation of the strains in artificial environments.
Growth temperatures vary between 6 and 37° with an optimum at 30°. At 6° strains untrained to grow at this temperature grow in about 5 days in Rosenow medium. Trained stains have a lag phase of 24 h. Growth curves show a longer logarithmic phase at 6° than at 30°. At the end of this phase the bacterial count is the same at both temperatures. Cl. carnofoetidum var. amylolyticum is, therefore, a true psychrotrophic species.
Glucose and pyruvate breakdown studied by Warburg's method confirms the slowing down of metabolism at 6°. A study of amino acid degradation indicates that serine is the substance most rapidly attacked at 37°, but at 6° the activity against serine disappears completely.
In Rosenow medium at the first inoculation 8 strains of Cl. perfringens out of 24 failed to grow at 6° after 30 days. They were all enterotoxic types. Among the other 16 strains none survived beyond the tenth transfer at 6° and it was not possible to train any strain to grow at this temperature. An examination of the growth curves shows that they are not truly psychrotrophic.     

Temperature-mediated dynamics of planktonic food chains: the effect of an invertebrate carnivore

1. This study involves an examination of two- and three-trophic-level food chains at two temperatures (18 and 25 °C) in order to determine how the addition of a carnivore to a predator–prey system can alter the dynamics of populations and how this effect may be temperature mediated. The system consisted of phytoplankton, Daphnia pulex and the flatworm Mesostoma ehrenbergii .
2. Although the plant–herbivore system is inherently unstable at 25 °C, the addition of the carnivore led to a further destabilization of the Daphnia –algal dynamics at the higher temperature. No destabilization effect of the carnivore was noted at 18 °C. At the lower temperature, all populations persisted and the carnivore induced changes only in the age structure of the Daphnia populations rather than in overall biomass.
3. The differential effects of the carnivore at two temperatures can be attributed to shifts in the life history, physiological rates and the reproductive strategy employed by Mesostoma .
4. Previous theoretical work has predicted that the addition of a third trophic level to an unstable predator–prey system should stabilize dynamics. Our results indicate that the effect of a carnivore on plant–herbivore dynamics can be significantly affected by ambient temperature.     

Temperature-mediated dynamics of planktonic food chains: the effect of an invertebrate carnivore

1. This study involves an examination of two- and three-trophic-level food chains at two temperatures (18 and 25 °C) in order to determine how the addition of a carnivore to a predator–prey system can alter the dynamics of populations and how this effect may be temperature mediated. The system consisted of phytoplankton, Daphnia pulex and the flatworm Mesostoma ehrenbergii .
2. Although the plant–herbivore system is inherently unstable at 25 °C, the addition of the carnivore led to a further destabilization of the Daphnia –algal dynamics at the higher temperature. No destabilization effect of the carnivore was noted at 18 °C. At the lower temperature, all populations persisted and the carnivore induced changes only in the age structure of the Daphnia populations rather than in overall biomass.
3. The differential effects of the carnivore at two temperatures can be attributed to shifts in the life history, physiological rates and the reproductive strategy employed by Mesostoma .
4. Previous theoretical work has predicted that the addition of a third trophic level to an unstable predator–prey system should stabilize dynamics. Our results indicate that the effect of a carnivore on plant–herbivore dynamics can be significantly affected by ambient temperature.     

Coexistence of similar genotypes of Daphnia magna in intermittent populations: response to thermal stress

We investigated the diversity and thermal response of a fitness related trait, juvenile growth rate, in seasonal population samples of Daphnia magna from two temperate ponds. Both populations were intermittent, i.e. they disappeared from the water body and recolonized seasonally by hatching from resting eggs in the sediment.
Temporally isolated clones of Daphnia magna showed the typical asymmetric response for growth rate with temperature and a sharp decline after the maximum response at 26°C (TMR). There was no evidence for genetically adapted seasonal groups. Despite significant genetic variation among clones and for phenotypic plasticity (G×E interactions without genetic correlations), seasonal groups of clones showed no shift in TMR and mean temperature reaction norms were similar among groups and both populations. Heritabilities remained similar among temperatures despite a large increase in genetic variance at stressfully high temperatures of 29°C and 32°C, due to simultaneous increase in environmental variance. Further, heritabilities remained high among sample periods and were not eroded during several months of asexual reproduction.
Regular diapause, an intrinsic feature of intermittent Daphnia populations, may replace the need for physiological temperature adaptation and promote maintenance of diversity through phenotypic similarity by reducing the time over which competitive interactions occur. Such populations are unlikely to be directly affected by elevated temperatures. They have a large potential for phenotypic plasticity as their TMR is higher than the temperature normally encountered.     

Conditions affecting growth and enterotoxin production by Staphylococcus aureus on temperature-abused chicken meat

Growth of Staphylococcus aureus at 15°C, with and without addition of representative spoilage bacteria, was studied in cooked, whole chicken meat and chicken broth. In the absence of competitors, the organism grew better in broth culture than on whole meat, but multiplied more slowly in broth when other organisms were present, even from twice the previous level of inoculum. The presence of competitors had no marked effect on the growth of Staph. aureus on whole meat. Enterotoxin A was not produced at 15°C on either whole meat or in broth, and occurred at 20°C only in pure culture. At 30° and 37°C, toxin was produced whether or not competitors were present. Toxin production by Staph. aureus appeared to be influenced more by growth temperature than by bacterial competition.     

Temperature effects on bioconcentration of DDE by Daphnia

1. Lake temperatures vary with season, latitude, elevation and as a result of thermal pollution. In addition, lake temperatures may increase with global warming. Radiotracer experiments were conducted to determine the effects of temperature on the bioaccumulation of lipophilic organic contaminants by zooplankton. Daphnia pulex were exposed to ¹⁴C-labelled DDE, a stable metabolite of the organochlorine pesticide DDT, in particle-free water for 24 h. An increase in temperature from 5 to 25 °C resulted in a 314% increase in bioconcentration factor (the ratio of contaminant concentration in the organism to contaminant concentration in the water).
2. To mimic the fluctuating temperatures experienced by zooplankton during diel vertical migration, we conducted experiments in which animals were exposed to 25 °C for 12 h in the light, then 15 °C for 12 h in the dark. Exposure to this fluctuating temperature regime for 48 h resulted in a 27–33% increase in bioconcentration factor relative to a constant 20 °C control.
3. Live animals accumulated more than twice the amount of DDE than freshly killed animals, indicating that the activity of the organism is important in bioconcentration. This finding sheds light on the possible mechanisms for the increase in bioconcentration at higher constant temperatures. Daphnia pump more water through their branchial chambers at higher temperatures. Thus, if the thoracic limbs are an important site of contaminant uptake, then animals are exposed to more contaminant molecules at higher temperatures. Other possible mechanisms include changes in the thickness of the diffusive boundary layer and changes in cell membrane permeability.     

Temperature effects on bioconcentration of DDE by Daphnia

1. Lake temperatures vary with season, latitude, elevation and as a result of thermal pollution. In addition, lake temperatures may increase with global warming. Radiotracer experiments were conducted to determine the effects of temperature on the bioaccumulation of lipophilic organic contaminants by zooplankton. Daphnia pulex were exposed to ¹⁴C-labelled DDE, a stable metabolite of the organochlorine pesticide DDT, in particle-free water for 24 h. An increase in temperature from 5 to 25 °C resulted in a 314% increase in bioconcentration factor (the ratio of contaminant concentration in the organism to contaminant concentration in the water).
2. To mimic the fluctuating temperatures experienced by zooplankton during diel vertical migration, we conducted experiments in which animals were exposed to 25 °C for 12 h in the light, then 15 °C for 12 h in the dark. Exposure to this fluctuating temperature regime for 48 h resulted in a 27–33% increase in bioconcentration factor relative to a constant 20 °C control.
3. Live animals accumulated more than twice the amount of DDE than freshly killed animals, indicating that the activity of the organism is important in bioconcentration. This finding sheds light on the possible mechanisms for the increase in bioconcentration at higher constant temperatures. Daphnia pump more water through their branchial chambers at higher temperatures. Thus, if the thoracic limbs are an important site of contaminant uptake, then animals are exposed to more contaminant molecules at higher temperatures. Other possible mechanisms include changes in the thickness of the diffusive boundary layer and changes in cell membrane permeability.     

Changes in food quality of the green alga Scenedesmus induced by Daphnia infochemicals: biochemical composition and morphology

1. The effects of Daphnia infochemicals on the morphology and biochemical composition of Scenedesmus were studied and subsequently their influence on Daphnia life history. Three species of Scenedesmus ( S. acutus, S. obliquus and S. subspicatus ) were tested for Daphnia -induced colony formation. Life history experiments were performed with Daphnia cucullata and D. magna feeding on unicellular or colonial S. acutus.
2. Colony formation was promoted when S. acutus and S. obliquus were exposed to filtered water from a D. magna culture. S. subspicatus did not form colonies when exposed to culture water, but showed some colony formation when exposed to live D. magna.
3. No clear differences were found in total lipid, protein or carbohydrate content between the unicellular and colonial ecomorphs of the three Scenedesmus species. However, fatty acid (FA) concentration and composition were changed. Total FA as a percentage of frozen dry weight (DW) were increased in colonies. The ratio of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) and the ratio of ω3 : ω6 fatty acids decreased in colonies.
4. A short-term grazing experiment with three size classes of Scenedesmus revealed lower clearance rates for small Daphnia when feeding on large colonial Scenedesmus.
5. Life history parameters of both Daphnia species were influenced by food type. The intrinsic rate of population increase ( r) was significantly lower for daphnids feeding on colonies.
6. The negative influence of colonial Scenedesmus on population growth of Daphnia seems to be determined by algal morphology rather than biochemical composition.     

Potato cold hardiness development and abscisic acid. II. De novo synthesis of proteins is required for the increase in free abscisic acid during potato (Solanum commersonii) cold acclimation

During cold acclimation of potato plantlets ( Solanum commersonii Dun, PI 458317), there are two transitory increases in free ABA content corresponding to a three-fold increase on the 2nd day and a five-fold increase on the 6th day (Ryu and Li 1993). During this period, plantlets increased in cold hardiness from −5°C (killing temperature, control grown at 22/18°C, day/night) to −10°C by the 7th day of exposure to 4/2°C (day/night). This increase in free ABA was not found when cycloheximide (CHI), an inhibitor of cytoplasmic protein synthesis, was added to the culture medium 6 h before exposure to low temperatures. Plantlets treated with CHI did not acclimate to cold, maintaining a hardiness level (−5°C) similar to that of the 22/18°C-grown plantlets. When the CHI-treated plantlets were exposed to low temperatures for 3 days, transferred to CHI-free culture medium and grown at low temperatures, the plantlets showed a transitory increase in free ABA 2 days later. This increase was followed by the development of cold hardiness (−8°C). Application of CHI to the culture medium after 3 days of cold acclimation, when the first ABA peak and a partial development of cold hardiness (−8°C) had occurred, blocked the second transitory increase in free ABA and resulted in no further development of cold hardiness. These results suggest that de novo synthesis of proteins is required for these transitory increases in free ABA during cold acclimation of potato plantlets.     

The recovery of heat-stressed Escherichia coli in lake water microcosms

C.-H. LIM AND K.P. FLINT. 1995. Escherichia coli was heat stressed at 55, 60 or 65°C in sterile flasks of lake water. After 6 h at these temperatures the viable count on nutrient agar had dropped below the limits of detection (1 colony in 100 ml). The flasks were transferred to a 15°C incubator and left for 7 d. Recovery of the stressed E. coli was shown to occur within 48 h at this temperature. Recovery also occurred in microcosms amended with 5o (v/v) synthetic sewage. The stressed E. coli multiplied in the amended but not in the unamended microcosms.     

Ventilation frequency compensation responses of three eurythermal estuarine fish exposed to moderate temperature increases

Ventilation frequency patterns of Trinectes maculatus, Morone americana and Leiostomus xanthurus were used to evaluate potential thermal stress after exposure to moderate temperature increases. Fish acclimated to 5°, 15° and 25° C were exposed to a 5°C T; fish acclimated to 30° C were exposed to a 2.5° C A T. Ventilation frequencies were measured at each acclimation temperature before the fishes were exposed over a 15-min period to the increased temperatures. Ventilation rates were then measured at the elevated temperatures for the next 24 h. Significant increases in rate frequency occurred after the temperature increases in T. maculatus and M. Americana acclimated to 5°, 15° and 25° C and in L. xanthurus acclimated to 15°, 25° and 30°C. In general, rate frequencies increased as the temperature increased. Ventilation rates stabilized quickly at the higher temperatures and remained relatively constant throughout the remaining exposure period. Acclimated rate-temperatures curves (R- T curves), acute R- T curves and Q10 temperature coefficients used to assess the significance of the changes in rate frequency and to compare the species in an ecologically meaningful way, showed that several adaptive types occurred among and between species. The Q,10's of the acute R- T curves, in most cases, were found to approximate those values derived for the acclimated R-T curve. This suggests that the temperature increases had a negligible effect, that is, little or no thermal stress occurred.     

Cold-tolerant (psychrotrophic) moulds and blue stain fungi from softwood in Sweden. Growth rates in relation to pH and temperature

Seventy isolates of moulds and blue stain fungi ( Zygomycota and Deuteromycota ), isolated from discoloured outdoor softwood in Sweden, comprising of 27 different species, (the two largest genera Penicillium and Cladosporium ) were investigated for their linear growth at three different start-pH values (5, 7 and 9.5) at two temperatures (2°C and 24°C) on malt extract agar (MEA). At 24°C all isolates showed growth at all three start-pH values except for one isolate which did not grow at initial-pH 9.5. After 21 days at 2°C at the three start pH-values, only six isolates showed no growth indicating that 64 of the isolates were cold-tolerant (psychrotrophic). Of these 64 strains, 58 showed growth at an initial pH of 9.5. Lower pH optima at 2°C than at 24°C were found for most of the isolates. The reduction of the linear growth at initial pH 9.5 in relation to the growth at optimal pH was more pronounced (higher) at the low temperature.     

Rooting of etiolated stem segments of Populus robusta – interaction of temperature, catechol and sucrose in the presence of IAA

Etiolated stem segments of Populus robusta Schneid. were cultured in test solutions each containing 2.0 mg/1 of IAA in combination with varying concentrations of catechol or sucrose or both at 10 ± 2°C, 25 ± 1°C and 35 ± 1°C in the dark. Cuttings did not root in catechol in the presence of IAA alone at 10 ± 2°C. There was no mortality in this culture solution at this temperature. However, cuttings rooted in this culture solution at 25 ± 1°C and 35 ± 1°C and also showed mortality, which was more severe at 35 ± 1°C than at 25 ± 1°C and more severe at higher than at lower concentrations of catechol. Rooting occurred on cuttings in catechol in the presence of IAA and sucrose even at 10 ± 2°C and was strongly promoted at 25 ± 1°C and 35 ± 1°C. The mortality of segments caused by catechol was markedly lowered in the presence of sucrose at these temperatures. Sucrose thus antagonised the toxic effect of catechol.     

Relation between Multiplication Rate and Temperature in Tetrahymena pyriformis, Strains HS and GL

SUMMARY. The multiplication rate of Tetrahymena pyriformis HS in proteose peptone medium was measured at 12 temperatures between 18.4°C. and 36.6°C. At the temperature optimum, 32.5°C., the generation time is 2.25 hours. The upper lethal temperature lies between 36.6°C. and 38.0°C. Similarly, a study of Tetrahymena pyriformis GL revealed a temperature optimum for multiplication of 29°C. with a generation time of 3.70 hours. The upper lethal temperature falls between 34.6°C. and 35.4°C. At all temperatures employed the HS strain of organisms multiplies more rapidly than strain GL. Under identical conditions, the two strains have distinctly different growth optima, upper lethal temperatures and growth rates.
As measured by multiplication rate the readjustment to a sudden change in temperature (from 18.4°C. to 27.7°C.) is completed very rapidly, with an effective lag time of about 1 hour. Such a shift in temperature gives rise to a small degree of division synchrony during the first and second population doublings which follow. Subsequently, all traces of division synchrony are lost.     

The effects of different soil water potentials, temperature and salinity on germination of seeds of the desert shrub Zygophyllum dumosum

The effects of different soil water potentials, temperature and NaCl concentration on seed germination of Zygophyllum dumosum Boiss., a common shrub in Israeli deserts, was investigated. Seeds had to be exposed to constant field capacity conditions (–0.0316 MPa) for a minimal period of two days before germination could start. Maximal germination under such conditions occurred after four days or more. Under simulated conditions of gradual dehydration of the soil, seeds were inhibited either at low soil water potentials (–0.10 to –10.00 MPa) or at high ones (–0.002 to –0.0398 MPa). Germination of Z. dumosum was independent of temperature in the range of 10–25°C, but strongly inhibited at 30 and 35°C. At 20°C germination was inhibited by salinity of the medium but still occurred (0.5%) even at a concentration of 0.5 M NaCl.