Induced tolerance expressed as relaxed behavioural threat response in millimetre-sized aquatic organisms

Natural selection shapes behaviour in all organisms, but this is difficult to study in small, millimetre-sized, organisms. With novel labelling and tracking techniques, based on nanotechnology, we here show how behaviour in zooplankton (Daphnia magna) is affected by size, morphology and previous exposure to detrimental ultraviolet radiation (UVR). All individuals responded with immediate downward swimming to UVR exposure, but when released from the threat they rapidly returned to the surface. Large individuals swam faster and generally travelled longer distances than small individuals. Interestingly, individuals previously exposed to UVR (during several generations) showed a more relaxed response to UVR and travelled shorter total distances than those that were naive to UVR, suggesting induced tolerance to the threat. In addition, animals previously exposed to UVR also had smaller eyes than the naive ones, whereas UVR-protective melanin pigmentation of the animals was similar between populations. Finally, we show that smaller individuals have lower capacity to avoid UVR which could explain patterns in natural systems of lower migration amplitudes in small individuals. The ability to change behavioural patterns in response to a threat, in this case UVR, adds to our understanding of how organisms navigate in the ‘landscape of fear’, and this has important implications for individual fitness and for interaction strengths in biotic interactions.     

Behavioural,developmental and morphological responses of Chironomus gr. thummi larvae (Diptera,Nematocera) to aquatic pollution

Populations of Chironomus gr. thummi larvae from two differently polluted lowland streams (Dommei, high cadmium and zinc; Ijse, medium copper and organic xenobiotics) were screened for behavioural and morphological responses to pollution. Behaviours such as locomotion (swimming and looping), respiration movements (ventilation) and inactivity were quantified with impedance conversion technique. Chironomids from the Dommel were more active than larvae from Ijse. In Ijse, deformed larvae showed less emergence, less locomotion and more ventilation than non-deformed larvae. In Dommel, deformed and normal larvae were equally fit (behaviour, emergence).     

A simple trickle chamber for rearing aquatic invertebrates

A chamber for laboratory rearing of aquatic invertebrates is described. The animals are reared in plastic dishes, through which a recirculating water volume is passed. The design ensures identical water quality in a high number of parallels. The chamber is constructed for rearing the freshwater amphipod Gammarus pulex, but can be used in studies of growth, reproduction etc. of other aquatic invertebrates as well.     

Aliesterase- (Ali-E) activity in Daphnia magna straus as a parameter for exposure to parathion

In vivo exposure of Daphnia magna Straus to parathion in concentrations from 0.05 to 5 µg/l resulted in a concentration-dependent decrease in aliesterase activity of the homogenate of the animals. Decrease in this activity after 48 hours of exposure was maximal for concentrations > 1.5 µg/l and amounted to about 90% of the activity of the blank.     

Cyst ornamentation in aquatic invertebrates: a defence against egg-predation

Hydrobiologia - Eggs, including encysted embryos (cysts) of aquatic invertebrates may not only be thick-walled, but also provided with various external ornamentations such as spines and...     

Analysis of factors controlling responses of an aquatic microcosm to organic loading

The changes in the numbers of individuals of the constituent species and ammonia concentrations in 6 subsystems (Bacteria, bacteria-Cyclidium, bacteria-Aeolosoma, bacteria-Chlorella, bacteria-Cyclidium-Chlorella, bacteria-Aeolosoma-Chlorella) of an aquatic microcosm were examined after the addition of organic substance (peptone) at both the young and mature stages of succession. Organic loading led to decreases in the numbers of bacteria and Aeolosoma after temporary increases in their numbers and an oscillation in the numbers of Cyclidium but the densities of these populations were maintained at constant values in the presence of Chlorella. When organic substance was added at a young stage where the photosynthetic activities of algae were high, invertebrates, especially Aeolosoma, were damaged by unionized ammonia which was formed as the pH value increased. In contrast, organic loading at the mature stage enhanced the population densities and biomasses of all constituent species.     

Heat shock as inducer of metamorphosis in marine invertebrates

Summary In most sessile marine invertebrates, metamorphosis is dependent on environmental cues. Here we report that heat stress is capable of inducing metamorphosis in the hydroid Hydractinia echinata. The onset of heat-induced metamorphosis is correlated with the appearance of heat-shock proteins. Larvae treated with the metamorphosis-inducing agents Cs⁺ or NH₄ ⁺ also synthesize heat-shock proteins. In heat-shocked larvae, the internal NH₄ ⁺-concentration increases. This fits the hypothesis that methylation plays a central role in control of metamorphosis. In the tunicate Ciona intestinalis, a heat shock is able to induce metamorphosis too. Offprint requests to: M. Walther     

Lipiniella arenicola (Chironomidae) compared withChironomus muratensis andCh. nudiventris: Distribution patterns related to depth and sediment characteristics,diet, and behavioural response to reduced oxygen concentrations

Density patterns ofLipiniella arenicola andChironomus muratensis were studied in Lake Wolderwijd (The Netherlands) using geostatistics, and compared with patterns of depth and silt contents of the sediment.L. arenicola was restricted to shallow (depth <1 m), sandy silt (silt content <5%), wind-exposed sediments.Ch. muratensis occurred all over the lake, but the highest densities were found in the sheltered silty North-West corner. Gut analyses of IV-instar larvae from the Ventjagers flats (Haringvliet, Lower Rhine-Meuse) showed thatL. arenicola fed by selectively grazing on benthic algae,Ch. muratensis by filtering and non-selective grazing, andCh. nudiventris by non-selective grazing. In the laboratory, behavioural responses to reduced oxygen concentrations were tested with the impedance conversion technique.L. arenicola was more sensitive to low oxygen concentrations (<3 mg l^–1) than bothChironomus species. It is argued that the differences in resistance to hypoxia and feeding behaviour are in accordance with the differences in distribution patterns. The restriction ofL. arenicola to the pure sand habitats is not attributable to an especially high sensitivity to low dissolved oxygen concentrations alone. Several behavioural adaptations enableL. arenicola larvae to live in the shallow, wind-exposed pure-sand habitats: the ability to select food items, tube strength, and site selection of ovipositing females or planktonic larvae.     

Electrophysiological and behavioural responses of mosquitoes to volatiles of Silene otites (Caryophyllaceae)

In order to understand the biological significance of flower odour for attraction of mosquitoes, electrophysiological responses to headspace flower odour samples of Silene otites (L.) Wibel were investigated on Culex pipiens pipiens biotype molestus Forskal 1775 and Aedes aegypti L. using coupled gas chromatographic-electroantennographic detection (GC-EAD). No remarkable differences in antennal responses to the odour compounds have been found between these two mosquito species. Further, the behavioural attractiveness of the electrophysiologically active compounds, singly or as multiple odour mixtures, was evaluated with bioassay experiments with C. pipiens molestus. In bioassays, C. pipiens responded to 14 electrophysiologically active compounds in different magnitudes (65–20%) and acetophenone, linalool oxide (pyranoid), phenyl acetaldehyde and phenylethyl alcohol were found as more attractive in comparison to the least attractive compound, hexanol. In two-stimulus choice test, mosquitoes were significantly more attracted to the mixture of the four most attractive compounds compared to the mixture of all 14 compounds. The results of present study confirm that floral odours are attractive cues for mosquitoes.     

Occurrence and antagonistic potential of Stenotrophomonas strains isolated from deep-sea invertebrates

Stenotrophomonas maltophilia is known to be of significance as opportunistic pathogen as well as a source of biocontrol and bioremediation activities. S. maltophilia strains have been isolated from rhizospheres, soil, clinical material, aquatic habitats, but little is known about Stenotrophomonas strains recovered from marine environments. During a survey of the biodiversity of Pseudomonas-like bacteria associated with deep-sea invertebrates six Stenotrophomonas strains were isolated from sponge, sea urchin, and ophiura specimens collected from differing Pacific areas, including the Philippine Sea, the Fiji Sea and the Bering Sea. 16S rRNA gene sequence analysis confirmed an assignment of marine isolates to the genus Stenotrophomonas as it placed four strains into the S. maltophilia CIP 60.77^T cluster and two related to the S. rhizophila DSM 14405^T. Together with a number of common characteristics typical of S. maltophilia and S. rhizophila marine isolates exhibited differences in pigmentation, a NaCl tolerance, a range of temperatures, which supported their growth, substrate utilization pattern, and antibiotics resistance. Strains displayed hemolytic and remarkable inhibitory activity against a number of fungal cultures and Gram-positive microorganisms, but very weak or none against Candida albicans. This is the first report on isolation, taxonomic characterization and antimicrobial activity of Stenotrophomonas strains isolated from deep-sea invertebrates.     

Population responses of plant-associated invertebrates to foraging by largemouth bass fry (Micropterus salmoides)

The effect of foraging by largemouth bass fry (Micropterus salmoides) upon invertebrates associated with aquatic macrophytes was determined using six 4 m² exclosures in Cochran Lake, Michigan during June 1978. The cladoceranSida crystallina rapidly declined in exclosures with fry, but increased in control exclosures without fry. Chironomids and chydorids showed little change in the exclosures.Invertebrate populations and foraging by fry were also monitored in the lake during 1976 and 1979.Sida declined rapidly in June of both years. In 1979, a decline from 2.2 × 10³ to 0.3 × 10³ individuals per m² and a sharp drop in the proportion of adults between 19 and 22 June coincided with the entry of a large school of fry into the study site on 19 June. Chironomids also declined during June of both years, while chydorids became increasingly abundant in mid-summer and showed no evidence of depletion by the fish.These results suggest that populations of certain prey, which are found locally in high densities in the littoral zone, may be highly susceptible to brief episodes of intense predation by fish fry.     

Efficacy of seed-based media for the mould-yeast conversion of Blastomyces dermatitidis

The efficacy of 20 seed-based media is reported for the in vitro mould-yeast conversion of Blastomyces dermatitidis, employing pharmamedia agar, peptone glucose agar, glucose agar and water agar as controls. The mould-yeast conversion varied significantly according to the culture medium, fungal strains and incubation period (p<0.01). Garden-pea, chick-pea, cow-pea, soyabean, peanut, green gram, French bean, lentil, okra and cottonseed converted all of the 7 B. dermatitidis test strains after 5 days of incubation at 37 ° C. Although the efficacy of many of these seed media was found to be at par with pharmamedia agar — a commercial cottonseed embryo-derived protein, garden-pea seed agar is adopted because of the wider availability and low fat content of this seed. The recommended composition of the medium comprises 2% aqueous seed extract, 2% glucose and pH 6–7. Only nigerseed and sunflower seeds failed to support the conversion of B. dermatitidis. Of the control media, peptone glucose agar, glucose agar and water agar did not support the conversion of 2 of the B. dermatitidis test strains. The mechanism underlying variable mould-yeast conversion of B. dermatitidis on seed-based media is not clearly understood. However, most of the seeds supporting excellent mould-yeast conversion are known for their high protein content. The conversion was apparently not affected by the fat content of the seeds or by incorporation of glucose in the medium.     

Development of a bioassay reagent usingPhotobacterium phosphoreum as a test for the detection of aquatic toxicants

Lyophilized cells ofPhotobacterium phosphoreum, rehydrated in 2% (w/v) NaCl in 0.022M KH₂PO₄ at pH 7.0, were used for developing an assay to test the acute toxicity of organic and inorganic compounds. The standardized assay gave good reproducibility of results with 11 organic and four inorganic compounds. Results were compared with reported data obtained with other test organisms and are within their sensitivity ranges. Environmental screening of wastes from oil and petrochemical industries is discussed.     

Analysis of Root Growth by Impedance Spectroscopy (EIS)

Electrical impedance spectroscopy (EIS) is investigated as a non-destructive method for monitoring root growth of tomato. This paper aims to (i) review the basic principles of EIS applied to the characterisation of the different parts of the soil–root–stem-electrode continuum, (ii) experiment the validity of the relationship between root weight and root capacitance taking into account the influence of the soil and plant electrodes position, (iii) describe an EIS analysis of the root growth of tomato plants. All experiments were carried out in 50 dm³ containers either in hydroponics at 930 μS for the test of root fresh or dry weight and root capacitance relationships, or in a potting mix (oxisol) for electrode placement tests and EIS estimation of root growth. Electrical measurements of the soil–root–stem-electrode continuum were done with a two-electrode measuring system using unpolarisable Ag–AgCl electrodes. A ‘root cutting’ and a ‘progressively immersed root system’ experiments were carried out in order to validate the relationship between root capacitance and root mass at 1 kHz. The effects of soil electrode and plant electrode placement were also tested, pointing out the sensitivity of the method to the insertion height of the “plant electrode” into the stem. For the root growth experiment, Impedance Spectra (IS) measurements were made just before harvesting the roots for dry weight and length determination. Measurements were made 14, 22, 26 and 39 days after planting, until flowering. The IS of the soil–root–stem-electrode continuum was modelled by a lumped electric circuit consisting of a series resistor R ₀ for the soil and of four parallel resistance (R _i )-capacitance (C _i ) circuits for the other components of the circuit. The model had nine parameters whose values were estimated by Complex Nonlinear Least Squares curve fitting. A stepwise ascendant regression was used to identify the electrical parameters that better correlated with root dry mass or length increment: C ₃ and C ₄ were well correlated with root dry mass with a r ² of 0.975, whereas root length was explained by the combination of 1/R ₃, C ₃, 1/R ₂ and 1/R ₁ with a r ² of 0.986. This work may be considered as a new methodological contribution to the understanding of root electrical properties in the non-destructive diagnosis of root systems.     

The relationship between temperature,competition and the potential for colonization of a subtropical pond by Daphnia magna

The role that temperature plays in excluding large daphnid species from subtropical and tropical ponds, and competitive relationships between Daphnia laevis and Daphnia magna, are explored. D. magna, a large temperate species is probably not excluded from subtropical or tropical systems by elevated temperatures. However, D. magna was a poorer competitor, under a restricted set of laboratory conditions, than D. laevis, the only pond dwelling daphnid in subtropical Florida. Competition is proposed as one mechanism that may limit the number of daphnid species in subtropical and tropical ponds and lakes. Reduced environmental fluctuations in subtropical and tropical systems (compared with temperate systems) may allow zooplankton populations to reach an environmental carrying capacity where competition limits the number of similar, coexisting species in a habitat.     

Larval aquatic and terrestrial mites infesting a temperate assemblage of mosquitoes

We collected 22,769 adult female mosquitoes, representing 27 species, from light traps in Norfolk, Virginia (2006–2007) and examined them to assess infestation by larval mites. Mosquitoes were parasitized by two species of aquatic (Acari: Arrenuridae: Arrenurus) and three species of terrestrial mites (Acari: Erythraeidae). The prevalence of infestation varied from 0.55% (2006) to 0.17% (2007). The mean intensity of parasitism ranged from 3.6 mites per host (2006) to 1.8 mites per host (2007). The most common host species for aquatic mites was Culex erraticus, while the most common host for terrestrial mites was Anopheles quadrimaculatus. Relationships between biotic and abiotic factors were investigated in an attempt to provide insight into temporal, spatial, and interspecific variation in mite–mosquito interactions. Scanning electron microscopy was used to examine the mode of attachment for larval mites. While the prevalence of aquatic mite parasitism was correlated for Culex erraticus, the invasive mosquito, Aedes albopictus, was never parasitized through the duration of the study. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The relationship between cladoceran body size and the growth of underyearling roach (Rutilus rutilus) (L.) in two shallow lowland lakes: a mechanism for density-dependent reductions in growth

Large planktonic Cladocera are typically the most important components of the diet of underyearling roach. Selection for large species and individuals by fish can result in a shift in the species composition of the cladoceran community as well as a reduction in the mean size of the individuals of large species and in the assemblage as a whole. Laboratory experiments demonstrated that underyearlings feeding on Daphnia hyalina smaller than 1 mm in length has a significantly lower intake of prey volume per unit time than when feeding on prey greater than 1.5 mm. A decrease in the nutritional quality of zooplankton prey, brought about by increasing predation pressure is suggested as the mechanism for density-dependent reductions in the growth of underyearling roach in eutrophic water bodies.     

A study of PstB cells during Dictyostelium migration and culmination reveals a unidirectional cell type conversion process

Summary The prestalk region of the Dictyostelium slug has recently been shown by Williams and his collaborators to consist of two distinct cell types, pstA and pstB cells. Here the movement of these cells in both the slug and culmination stages has been examined with the use of vital dyes. In the slug some of the pstB cells are continually lost from the prestalk region as small clusters of cells. These cells move through the prespore region and temporarily lie in the rearguard region at the posterior end of the slug. They are finally left in the slug's slime track as single cells or groups of a few cells. When culmination is initiated the pstB cells move as a whole from the prestalk region to the base where they join the rearguard cells to form the basal disc of the fruiting body. Transplantation experiments reveal that the rearguard cells form an outer ring portion of the basal disc and the pstB cells form an inner portion to which the stalk attaches. The continuous loss of one cell type during the slug stage without any change in cell type proportions suggests that cell types are redifferentiating. Grafting and transplantation experiments reveal that there is a unidirectional flow of cells through successive steps of cell type conversion. Prespore cells redifferentiate as anterior-like cells which migrate to the prestalk region and become pstA cells. The pstA cells then replace the pstB cells that are lost from the slug.