Role of melatonin in the control of depth distribution of <Emphasis Type="Italic">Daphnia magna</Emphasis>

Previous studies confirmed the presence of melatonin in Daphnia magna and demonstrated diurnal fluctuations in its concentration. It is also known that in several invertebrate species, melatonin affects locomotor activity. We tested the hypothesis that this hormone is involved in the regulation of Daphnia diel vertical migration (DVM) behaviour that is well recognized as the adaptive response to predation threat. Using ‘plankton organs’, we studied the effect of three concentrations of exogenous melatonin (10⁻⁵, 10⁻⁷, 10⁻⁹ M) on DVM of both female and male D. magna in the presence or absence of chemical cue (kairomone) of planktivorous fish. Depth distribution was measured six times a day, using infrared-sensitive closed circuit television cameras. Our results showed a significant effect of melatonin on the mean depth of experimental populations, both males and females, but only when melatonin was combined with fish kairomone. Females stayed, on average, closer to the surface than males, both responding to the presence of kairomone by descending to deeper strata. In the presence of exogenous melatonin and with the threat of predation, Daphnia stayed closer to the surface and their distribution was more variable than that of individuals, which were exposed to the kairomone alone. Approaching the surface in the presence of predation threat seems to be maladaptive. We postulate the role of melatonin as a stress signal inhibitor in molecular pathways of response to predation threat in Cladocera.     

Impact of food concentration on diel vertical migration behaviour of <Emphasis Type="Italic">Daphnia pulex</Emphasis> under fish predation risk

Vertical migration of Daphnia represents the best-studied predator-avoidance behaviour known; yet the mechanisms underlying the choice to migrate require further investigation to understand the role of environmental context. To investigate the optimal habitat choice of Daphnia under fish predation pressure, first, we selected the individuals exhibiting strong migration behaviour. The animals collected from the hypolimnion during the daytime were significantly larger, being more conspicuous, and in turn performed stronger diel vertical migration (DVM) when exposed to fish cue. We called them strong migrants. Second, we provided the strong migrant D. pulex with food at high and intermediate (1 and 0.4 mg C l⁻¹, respectively) levels, which were well above the incipient limiting level and of high quality. They traded the benefits of staying in the warm water layer and moved down to the cold water in response to fish cue indicating fish predation. The availability of food allowed the animals to stay in the cold hypolimnion. However, at the low food level (0.1 mg C l⁻¹), which is an additional constraint on fitness, Daphnia moved away from the cold hypolimnion. Poor food condition resulted in strong migrant Daphnia to cease migration and remain in the upper warmer water layer. Although temperature is known to be a more important cost factor of DVM than food, our results clearly show that this is only true as long as food is available. It becomes clear that food availability is controlling the direction of vertical positioning when daphnids experience a dilemma between optimising temperature and food condition while being exposed to fish cue. Then they overlook the predation risk. Thus, the optimal habitat choice of Daphnia appears to be a function of several variables including temperature, food levels and fish predation. Handling editor: S. I. Dodson     

Clutch size and body size at first reproduction in Daphnia pulicaria at different levels of food and predation

Field data from seven alpine lakes in Serra da Estrela. Portugal.show that reproduction in Daphnia may be as efficiently controlledby fish predation and copepod predation on eggs in brood cavitiesas it is by food limitation. Body length and clutch size estimatesin Daphnia pulicaria revealed high inter- and intra-populationvariability in maturation size (body size at first reproduction).and in number of eggs per clutch. Daphnia at first maturationin lakes stocked with rainbow trout were half the size of thosefound in fishless lakes (body length of 0.86–0.95 and1.55–1.81 mm. respectively). The mean number of eggs perclutch was reduced to a similar degree by food limitation, predationby fish and copepod predation on eggs in brood cavities, butthe underlying mechanisms of this reduction were different.Food limitation caused smaller clutch sizes in all individuals,so variation remained the same. Fish predation caused the selectiveremoval of individuals with maximum clutches, so variation decreased.Copepod predation caused removal of eggs from brood cavitiesof randomly infested females, so that variation increased, particularlyat a high food level when non-infested females carried largeclutches of eggs.     

Goose-mediated nutrient enrichment and planktonic grazer control in arctic freshwater ponds

A dramatic increase in the breeding population of geese has occurred over the past few decades at Svalbard. This may strongly impact the fragile ecosystems of the Arctic tundra because many of the ultra-oligotrophic freshwater systems experience enrichment from goose feces. We surveyed 21 shallow tundra ponds along a gradient of nutrient enrichment based on exposure to geese. Concentrations of total phosphorus (P) and dissolved inorganic nitrogen (DIN) in the tundra ponds ranged from 2–76 to 2–23 μg l⁻¹ respectively, yet there was no significant increase in phytoplankton biomass (measured as chlorophyll a; range: 0.6–7.3 μg l⁻¹) along the nutrient gradient. This lack of response may be the result of the trophic structure of these ecosystems, which consists of only a two-trophic level food chain with high biomasses of the efficient zooplankton grazer Daphnia in the absence of fish and scarcity of invertebrate predators. Our results indicate that this may cause a highly efficient grazing control of phytoplankton in all ponds, supported by the fact that large fractions of the nutrient pools were bound in zooplankton biomass. The median percentage of Daphnia–N and Daphnia–P content to particulate (sestonic) N and P was 338 and 3009%, respectively, which is extremely high compared to temperate lakes. Our data suggest that Daphnia in shallow arctic ponds is heavily subsidized by major inputs of energy from other food sources (bacteria, benthic biofilm), which may be crucial to the persistence of strong top–down control of pelagic algae by Daphnia.     

Early responses of plankton and turbidity to biomanipulation in a shallow prairie lake

We evaluated the effect of a fish removal from a shallow, turbid, eutrophic lake. By late May (following an October fish removal), the cladoceran community shifted from small-bodiedBosmina andChydorus (less than 100 l⁻¹) to largerDaphnia (over 100 l⁻¹). During the periods of peak daphnid abundance (late May–June) chlorophyll-a concentrations and edible diatoms were reduced and water transparency improved dramatically. Total phosphorus was not significantly lowered during this period. Although this clear-water phase was short-lived (May, June and early July), it corresponded to the critical period of plant growth and allowed dramatic increases in submergent macrophytes.     

Spatial variability in the abundance of pelagic invertebrate predators in relation to depth and turbidity

The abundance of pelagic invertebrate predators in relation to turbidity and depth gradients in Lake Hiidenvesi (southern Finland) were studied. In the shallow (<5 m) and the most turbid (up to 75 NTU) part of the lake, the community of invertebrate predators consisted of cyclopoid copepods (max biomass >500 μg dw l⁻¹) and Leptodora kindtiii (Focke) (17 μg dw l⁻¹), while in the less turbid (10–40 NTU) stratifying area Chaoborus flavicans (Meigen) dominated (max 146 μg dw l⁻¹). In the temporarily stratifying and moderately turbid basin Chaoborus and small-bodied invertebrate predators co-existed. Mysis relicta (Lovén) occurred only in the stratifying area (max 15 μg dw l⁻¹). The results suggested that both water depth and turbidity contributed to the community structure of Chaoborus flavicans. Depth great enough for stratification was of special importance and its effect was amplified by elevated turbidity, while high turbidity alone could not maintain chaoborid populations. Mysis relicta also requires a hypolimnetic refuge but is more sensitive to low oxygen concentrations and may therefore be forced to the epilimnion where it is vulnerable to fish predation. Cyclopoids as rapid swimmers can take advantage at elevated turbidity levels and coexist in high biomass with fish even in shallow water. Leptodora kindtii can form high biomass despite planktivorous fish providing that turbidity exceeds 20 NTU. The results demonstrated that depth and water turbidity can strongly regulate the abundance and species composition of invertebrate predators. These factors must thus be taken into account when applying food web management, which aims to reduce phytoplankton biomass by depressing planktivorous fish.     

Heterotrophic production of eicosapentaenoid acid by the diatom Nitzschia laevis: effects of silicate and glucose

The effects of silicate and glucose on growth and eicosapentaenoic acid (EPA) production by the diatom Nitzschia laevis were studied. By alternately altering the concentrations of silicate (2.7–64 mg l⁻¹) and glucose (1–40 g l⁻¹) in the medium, the highest cell dry weight (ca. 5.5 g l⁻¹) was obtained at 20 g l⁻¹ glucose and 32 mg l⁻¹ silicate, while the highest specific growth rate (ca. 0.65 day⁻¹) was obtained at a relatively low glucose concentration (5 g l⁻¹) and high silicate concentrations (32–64 mg l⁻¹). At glucose levels of 5 and 20 g l⁻¹, EPA content was higher with lower silicate concentrations (2.7 and 16 mg l⁻¹ silicate, respectively), while at a silicate level of 16 mg l⁻¹, higher glucose concentrations (20–40 g l⁻¹) facilitated EPA formation. The highest EPA yield (131 mg l⁻¹) was obtained at 20 g l⁻¹ glucose and 32 mg l⁻¹ silicate, while the highest EPA productivity (15.1 mg l⁻¹ day⁻¹) was obtained at 20 g l⁻¹ glucose and 64 mg l⁻¹ silicate. Journal of Industrial Microbiology & Biotechnology (2000) 25, 218–224. Received 08 May 2000/ Accepted in revised form 21 July 2000     

Contributions of Different Organic Carbon Sources to <Emphasis Type="Italic">Daphnia</Emphasis> in the Pelagic Foodweb of a Small Polyhumic Lake: Results from Mesocosm DI<Superscript>13</Superscript>C-Additions

Abstract Freshwater ecosystems derive organic carbon from both allochthonous and autochthonous sources. We studied the relative contributions of different carbon sources to zooplankton in a small, polyhumic, steeply stratified lake, using six replicate surface-to-sediment enclosures established during summer and autumn 2004. We added ¹³C-enriched bicarbonate to the epilimnion of half the enclosures for three weeks during each season and monitored carbon stable isotope ratios of DIC, DOC, POC and Daphnia, along with physical, chemical and biological variables. During summer, ¹³C-enriched DIC (δ¹³C up to 44 ± 7.2‰) was soon taken up by phytoplankton (δ¹³C up to −5.1 ± 13.6‰) and was transmitted to Daphnia (δ¹³C up to −1.7 ± 7.2‰), demonstrating consumption of phytoplankton. In contrast, during autumn, ¹³C-enriched DIC (δ¹³C up to 56.3 ± 9.8‰) was not transmitted to Daphnia, whose δ¹³C became progressively lower (δ¹³C down to −45.6 ± 3.3‰) concomitant with decreasing methane concentration. Outputs from a model suggested phytoplankton contributed 64–84% of Daphnia diet during summer, whereas a calculated pelagic carbon mass balance indicated only 30–40% could have come from phytoplankton. Although autumn primary production was negligible, zooplankton biomass persisted at the summer level. The model suggested methanotrophic bacteria contributed 64–87% of Daphnia diet during autumn, although the calculated carbon mass balance indicated a contribution of 37–112%. Thus methanotrophic bacteria could supply virtually all the carbon requirement of Daphnia during autumn in this lake. The strongly ¹³C-depleted Daphnia values, together with the outputs from the models and the calculated carbon mass balance showed that methanotrophic bacteria can be a greater carbon source for Daphnia in lakes than previously suspected.     

A reliable protocol for plant regeneration from callus culture of <Emphasis Type="Italic">Phalaenopsis</Emphasis>

Summary Callus of Phalaenopsis Nebula was induced from seed-derived protocorms on 1/2 Murashige and Skoog (MS) basal medium plus 0–1.0 mg l⁻¹ (0–4.52 μM) N-phenyl-N′-1,2,3,-thiadiazol-5-yl urea (TDZ) and/or 0–10 mg l⁻¹ (0–45.24 μ M) 2,4-dichlorophenoxyacetic acid (2,4-D). Protocorms 2 mo. old performed better than 1-mo.-old protocorms for callus induction. More calluses formed on 1/2 MS basal medium supplemented with 0.1–1.0 mg l⁻¹ (0.45–4.52 μM) TDZ. These calluses could be maintained by subculturing every month with basal medium supplemented with 0.5 mg l⁻¹ (2.27 μM) TDZ and 0.5 mg l⁻¹ (2.26 μM) 2,4-D. Protocorm-like bodies were formed, and plants regenerated from these calluses on 1/2 MS basal medium alone or supplemented with 0.1–1.0 mg l⁻¹ (0.45–4.52 μM) TDZ. Plantlets were then potted on sphagnum moss in the greenhouse and grew well. No chromosomal abnormalities were found among the root-tip samples of 21 of the regenerated plantlets that were successfully acclimatized.     

Laccase and Manganese Peroxidase Activities of Phellinus Robustus and Ganoderma Adspersum Grown on Food Industry Wastes in Submerged Fermentation

Phellinus robustus produced both laccase (700–4,000 U l⁻¹) and manganese peroxidase (MnP) (1,000–11,300 U l⁻¹) in fermentation of nine food wastes, whereas Ganoderma adspersum produced only laccase (600–34,000 U l⁻¹). Glucose provided high laccase and MnP activity of P. robustus but repressed enzyme production by G. adspersum. Ammonium sulphate and ammonium tartrate increased the P. robustus laccase yield (3-fold), whereas the accumulation of MnP was not enhanced by additional nitrogen.     

Can Simple Empirical Equations Describe the Seasonal Dynamics of Bacterioplankton in Lakes: An Eight-Year Study in Shallow Hypertrophic and Biologically Highly Dynamic Lake Søbygård,Denmark

Abstract Seasonal variation in bacterioplankton abundance, biomass, and bacterioplankton production was studied over eight years in hypertrophic Lake S?byg?rd. Biologically, the lake is highly variable; this is due mainly to large interannual variation in fish recruitment. Bacterioplankton production was low during winter, typically 1–3 × 10⁷ cells l⁻¹ h⁻¹, and high during summer, albeit greatly fluctuating with maximum rates typically ranging from 60 to 90 × 10⁷ cells l⁻¹ h⁻¹ (or 0.4 to 0.6 mg C l⁻¹ day⁻¹). Less pronounced variations were found in bacterioplankton abundance, which typically ranged from 3–8 × 10⁹ cells l⁻¹ in winter to 15–30 × 10⁹ cells l⁻¹ during summer. The specific growth rate of bacterioplankton varied from 0.02–0.2 d⁻¹ in winter to 0.5–2.3 day⁻¹ during summer. Interpolated mean bacterioplankton production, in terms of carbon, ranged from 0.08 to 0.16 mg C l⁻¹ day⁻¹, corresponding to 1.6–5.5% of the phytoplankton production, while biomass ranged from 0.28 to 0.36 mg C l⁻¹, corresponding to 1.9–4.6% of the phytoplankton biomass. We conducted regression analysis, relating the bacterioplankton variables to a number of environmental variables, and evaluated the interannual parameter variability. Chlorophyll a and phytoplankton production contributed less to the variation in the bacterioplankton variables than in most previous analyses using data from less eutrophic systems. We suggest that the proportion of phytoplankton production that is channelized through bacterioplankton in lakes decreases with increasing trophic state and decreasing mean depth. This probably reflects a concurrent increase in fish predation on macrozooplankton and loss by sedimentation. An important part of the residual variation in the equations hitherto proposed in the literature could be explained by variation in macrozooplankton biomass and pH > 10.2. A negative effect of high pH on bacterioplankton production was confirmed by laboratory experiments. The impact of different zooplankton varies considerably, with Daphnia seeming to have a negative impact on bacterioplankton abundance and, thereby, indirectly on bacterioplankton production, while Bosmina, rotifers, and cyclopoid copepods seem to stimulate both abundance and production. Bosmina apparently also stimulate the bacterioplankton specific growth rate. Received: 8 February 1996; Accepted: 16 July 1996     

Coexistence of two similar copepod species, <Emphasis Type="Italic">Eudiaptomus gracilis</Emphasis> and <Emphasis Type="Italic">E. graciloides</Emphasis>: the role of differential predator avoidance

Habitat choice in relation to environmental factors of two coexisting calanoid copepod species, Eudiaptomus gracilis and E. graciloides, was studied in a mesotrophic lake and in large indoor mesocosms. Both species and sexes showed pronounced diel vertical migration (DVM) in the field. In 12 m deep mesocosms with free ranging fish DVM was observed and species increased day depth over time. No changes were observed in copepod day depth over time in experiments with fish kairomone. It is hypothesized that fish kairomone acts as an early warning system to copepods which respond by moving deeper, but only as far as the thermocline. For full DVM, a nearby mechanical stimulus is necessary. Thus, as fish go deeper to feed, copepods retreat. The response of copepods to fish predation, in the presence of low and high numbers of Daphnia,shows that copepods effectively use Daphniaas living shields to avoid predation. The two species adopt different vertical migration strategies depending on whether there are high or low numbers of Daphniapresent. A dominant feature of mesocosm experiments was the night time aggregating (lekking) of E. gracilis males at the surface. When the spring and autumn percentages of risk takers in the epilimnion were compared, E. gracilis, particularly males, suffered the greatest cost.     

Behavioural and population responses to changing availability of <Emphasis Type="Italic">Artemia</Emphasis> prey by moulting black-necked grebes, <Emphasis Type="Italic">Podiceps nigricollis</Emphasis>

Chemical communication may inform about the location of prey, predators, co-specifics, and mate partners in zooplankton. In this study, we evaluated several life-history traits of the rotifer, Brachionus calyciflorus, exposed to conditioned media by a rotifer predator (Asplanchna brightwelli) and a cladocera competitor (Daphnia similis), quantifying population growth and life-table demography at two algal food levels (2.0 and 0.5 × 10⁶ cells ml⁻¹ of Chlorella pyrenoidosa). At both food levels, B. calyciflorus grown in predator-conditioned media had lower population abundance and slower population growth rate than controls. Conversely, the competitor-conditioned media treatments produced both higher rotifer population abundance and faster population growth rate than controls. Life-history parameters varied significantly depending on the presence of predator and competitor-conditioned media. The Asplanchna-conditioned media significantly decreased gross reproductive rate (GRR): 8–9 offsprings per female; net reproductive rate (R ₀): 6–7 offsprings per female; population growth rate (r): 0.34–0.37 day⁻¹; and increased generation time (T): 5.5–5.6 days. On the other hand, The Daphnia-conditioned media significantly increased the GRR (13–14 offsprings per female); net reproductive rate (8–9 offsprings per female); population growth rate (0.42–0.43 day⁻¹); and decreased generation time (4.9–5.0 days). However, the effects of food level on the life-history characteristic were not significant in both treatments. Maximum values of the population abundance and the population growth rate are significantly influenced by the predator densities and pre-culture time. This study suggests that rotifers use variable life-history strategies (low reproduction and high survivorship versus high reproduction and low survivorship) based on the presence of predators and competitors.     

<Emphasis Type="Italic">In vitro</Emphasis> selection of NaCl-tolerant callus lines and regeneration of plantlets in a bamboo (<Emphasis Type="Italic">Dendrocalamus strictus</Emphasis> Nees)

Summary Sodium chloride-tolerant plantlets of Dendrocalamus strictus were regenerated successfully from NaCl-tolerant embryogenic callus via somatic embryogenesis. The selection of embryogenic callus tolerant to 100 mM NaCl was made by exposing the callus to increasing (0–200 mM) concentrations of NaCl in Murashige and Skoog medium having 3% (w/v) sucrose, 0.8% (w/v) agar, 3.0 mg l⁻¹ (13.6 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5mg l⁻¹ (2.3μM) kinetin (callus initiation medium). The tolerance of the selected embryogenic callus to 100 mM NaCl was stable through three successive transfers on NaCl-free callus initiation medium. The tolerant embryogenic callus had high levels of Na⁺, sugar, free amino acids, and proline but a slight decline was recorded in K⁺ level. The stable 100 mM NaCl-tolerant embryogenic callus differentiated somatic embryos on maintenance medium [MS medium +3% sucrose +0.8% agar +2.0 mg l⁻¹ (9.0 μM) 2,4-D+0.5 mg l⁻¹ (2.3 μM) kinetin] supplemented with different (0–200 mM) concentrations of NaCl. About 39% of mature somatic embryos tolerant to 100 mM NaCl germinated and converted into plantlets in germination medium [half-strength MS+2% sucrose+0.02 mg l⁻¹ (0.1 μM) α-naphthaleneacetic acid +0.1 mg l⁻¹ (0.49 μM) indole-3-butyric acid] containing 100 mM NaCl. Of these plantlets about 31% established well on transplantation into a garden soil and sand (1:1) mixture containing 0.2% (w/w) NaCl.     

Mechanisms of vasodilation in the dorsal aorta of the elephant fish, <Emphasis Type="Italic">Callorhinchus milii</Emphasis> (Chimaeriformes: Holocephali)

This study investigated vasodilator mechanisms in the dorsal aorta of the elephant fish, Callorhinchus milii, using anatomical and physiological approaches. Nitric oxide synthase could only be located in the perivascular nerve fibres and not the endothelium of the dorsal aorta, using NADPH histochemistry and immunohistochemistry. In vitro organ bath experiments demonstrated that a NO/soluble guanylyl cyclase (GC) system appeared to be absent in the vascular smooth muscle, since the NO donors SNP (10⁻⁴ mol l⁻¹) and SIN-1 (10⁻⁵ mol l⁻¹) were without effect. Nicotine (3 × 10⁻⁴ mol l⁻¹) mediated a vasodilation that was not affected by ODQ (10⁻⁵ mol l⁻¹), l-NNA (10⁻⁴ mol l⁻¹), indomethacin (10⁻⁵ mol l⁻¹), or removal of the endothelium. In contrast, the voltage-gated sodium channel inhibitor, tetrodotoxin (10⁻⁵ mol l⁻¹), significantly decreased the dilation induced by nicotine, suggesting that it contained a neural component. Pre-incubation of the dorsal aorta with the calcitonin gene-related peptide (CGRP) receptor antagonist, CGRP_8–37 (10⁻⁶ mol l⁻¹) also caused a significant decrease in the nicotine-induced dilation. We propose that nicotine is mediating a neurally-derived vasodilation in the dorsal aorta that is independent of NO, prostaglandins and the endothelium, and partly mediated by CGRP.     

Spatial variation in sea urchins, fish predators, and bioerosion rates on coral reefs of Belize

Although sea urchins are critical for controlling macroalgae on heavily fished coral reefs, high densities threaten reefs, as urchins are also prodigous bioeroders. This study examined urchin population characteristics, bioerosion rates, their fish predators (Labridae), and potential competitors (Scaridae) on unprotected reefs and a reef within a marine protected area (MPA) in the lagoonal regions off Belize. Urchin density (<1 m⁻²) and bioerosion rates (∼0.2 kg CaCO₃ m⁻² year⁻¹) were lowest and members of the Labridae were the highest (∼20 fish 200 m⁻³) within the MPA, while several unprotected reefs had higher (∼18–40 m⁻²) urchin densities, lower Labridae abundances (1–3 fish 200 m⁻³), and bioerosion rates ranging from ∼0.3–2.6 kg CaCO₃ m⁻² year⁻¹. Urchin abundances were inversely related to Labridae (wrasses and hogfish) densities; however, on reef ridges, low algal cover (∼15%), small urchin size (∼14 mm), and low proportion of organic material in urchin guts suggested food limitation. Both top–down (predation) and bottom–up factors (food limitation) likely contribute to the control of urchins, predominantly Echinometra viridis, off Belize, thereby potentially diminishing the negative impacts of bioerosion activities by urchins.     

Predator-induced diapause in Daphnia magna may require two chemical cues

The production of diapausing eggs by Daphnia magna stimulated by fish exudates can be explained as an anti-predator defence ensuring genome protection in periods of high risk from fish predation. The combined effects on the induction of D. magna diapause of an “alarm” chemical originating from injured conspecific prey and fish kairomones were tested. The results of the experiment showed that the cues when present together promote both the production of ephippial eggs and male formation, indicating their role in the synchronization of the entire mode of Daphnia sexual reproduction. Ephippial eggs were only produced in the presence of both fish kairomone and conspecific alarm chemicals, while male offspring occurred in the treatments where both, one or none of the cues were present. However, production of males was the highest when both cues were provided. D. magna responded similarly to the tested cues whether or not the hypothetical alarm substance associated with predator odour came from Daphnia specimens actually eaten by fish or from crushed conspecific individuals. However, chemicals from crushed chironomid larvae combined with fish kairomones did not induce a similar response in D. magna. The relative advantage of utilization of alarm cues or predator kairomones in the induction of defence responses in prey organisms is discussed. Received: 8 June 1998 / Accepted: 11 January 1999     

Interclonal variation in diel horizontal migration behaviour of the water flea <Emphasis Type="Italic">Daphnia magna</Emphasis>—searching for a signature of adaptive evolution

In shallow temperate lakes, zooplankton populations may exhibit diel horizontal migration (DHM) and move towards macrophytes during the day to avoid fish. Using a natural Daphnia magna population, we undertook an experimental investigation aimed to describe the genetic variation for DHM and to study whether an adaptive micro-evolutionary response occurred to changes in macrophyte coverage and fish predation pressure through time. Twenty-seven D. magna clones were hatched from ephippia in the sediment of shallow Lake Ring, Denmark. This lake was eutrophied during the 20th century and was subject to restoration measures in the 1970s. The DHM behaviour of the clones was observed both in the presence and absence of fish kairomone. Significant interclonal variation in DHM behaviour occurred in both treatments. To study the micro-evolutionary response of the Lake Ring D. magna population, two approaches were used. First, we compared the DHM behaviour of clones derived from ephippia collected at different depths. A comparison was conducted between clones resurrected from the period of eutrophication (1960–1980) and from the period of recovery (1986–2000). A significant treatment (presence and absence of fish kairomone) × period interaction effect was identified, suggesting a significant micro-evolutionary response for DHM behaviour. The D. magna clones exhibited a significantly stronger horizontal migration response during the period of eutrophication than in the recovery phase. Second, clonal means, representing the influence of the genotype on the trait, were correlated with environmental conditions (macrophyte cover, fish predation pressure and Secchi depth). The results of this analysis also suggest that a micro-evolutionary response by Daphnia has occurred in reaction to changes in fish predation pressure. In periods with high fish predation pressure, Daphnia migrated more strongly towards the plants. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Optimization of polyhydroxybutyrate production by <Emphasis Type="Italic">Bacillus</Emphasis> sp. CFR 256 with corn steep liquor as a nitrogen source

Polyhydroxyalkanotes (PHAs), the eco-friendly biopolymers produced by many bacteria, are gaining importance in curtailing the environmental pollution by replacing the non-biodegradable plastics derived from petroleum. The present study was carried out to economize the polyhydroxybutyrate (PHB) production by optimizing the fermentation medium using corn steep liquor (CSL), a by-product of starch processing industry, as a cheap nitrogen source, by Bacillus sp. CFR 256. Response surface methodology (RSM) was used to optimize the fermentation medium using the variables such as corn steep liquor (5–25 g l⁻¹), Na₂HPO₄ 2H₂O (2.2–6.2 g l⁻¹), KH₂PO₄ (0.5–2.5 g l⁻¹), sucrose (5–55 g l⁻¹) and inoculum concentration (1–25 ml l⁻¹). Central composite rotatable design (CCRD) experiments were carried out to study the complex interactions of the variables. The optimum conditions for maximum PHB production were (g l⁻¹): CSL-25, Na₂HPO₄ 2H₂O-2.2, KH₂PO₄ − 0.5, sucrose − 55 and inoculum − 10 (ml l⁻¹). After 72 h of fermentation, the amount of PHA produced was 8.20 g l⁻¹ (51.20% of dry cell biomass). It is the first report on optimization of fermentation medium using CSL as a nitrogen source, for PHB production by Bacillus sp.     

Fish and nutrient enrichment effects on rotifers in a Mediterranean shallow lake: a mesocosm experiment

A mesocosm experiment was conducted “in situ” in a Chara dominated shallow lake near Valencia (Spain) to study top–down and bottom–up effects on rotifers by means of nutrient and fish additions. Both processes were important in determining rotifer abundance, biomass and diversity. A total of 36 mesocoms were established with triplicate treatment combinations of three fish levels (from no fish to 45 individuals of Gambusia holbrooki males) and four nutrient enrichment levels (from no additions to 10 mg l⁻¹ nitrate-N and 1 mg l⁻¹ phosphate-P). The main effect was a notable increase of planktonic and plant associated rotifers densities with fish. Rotifers benefited from mosquitofish predation on microcrustaceans and chironomids. The results showed a marked negative relationship between rotifer and cyclopoid abundances, indicating the importance of the predatory pressure of cyclopoids on rotifers. Effects on rotifer diversity were also evident, in general rotifer diversity decreased with nutrients and increased with fish. The effects of nutrients analysed at species level showed two contrasting density responses: an increase or a decrease with nutrients, which levelled off at high nutrient concentrations. High-level nutrient additions (from 5 mg l⁻¹ nitrate-N and 0.5 mg l⁻¹ phosphate-P) induced a switch to a turbid state with macrophyte disappearance. Most planktonic rotifer species, as well as plant associated ones, diminished when the turbid state was well established, especially in the mesocosms without fish. In the turbid mesocosms, relative abundance of plant-associated rotifers (as a whole) was higher than that of planktonic rotifers. The changes in rotifer species composition after the switch from a clear to a turbid water state are also described. Species of the genus Anuraeopsis, Trichocerca and Hexarthra, dominant in the clear water state, practically disappeared in the turbid water state, in which Proalides tentaculatus and Lecane nana were the main species. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research