Anthraquinones production in Rubia tinctorum cell suspension cultures: Down scale of shear effects

The effect of turbulence on suspended cells is one of the most complex problems in the scale-up of cell cultures. In the present paper, a direct comparison of the effects of turbulence on suspension cultures of Rubia tinctorum in a standard bioreactor and in shake flask cultures was done. A procedure derived from the well known global method proposed by Nishikawa et al. (1977) [39] was applied. Standard flasks and four-baffled shake flasks were used. The effect of turbulence and light irradiation on cell viability, biomass, and anthraquinones (AQs) production was evaluated. The biomass concentration and AQs production obtained using baffled shake flasks agitated at 360 rpm were similar to that achieved in R. tinctorum suspension cultures growing in a stirred tank bioreactor operating at 450 rpm, previously published (Busto et al., 2008 [17]). The effect of light on AQs production was found to be very significant, and a difference of up to 48% was found in cells with and without illumination after 7 days of culture. It is concluded that this down-scaled and simple flask culture system is a suitable and valid small scale instrument for the study of intracellular mechanisms of turbulence-induced AQs production in R. tinctorum suspension cultures.     

Turbulence and stratification in Priest Pot,a productive pond in a sheltered environment

Priest Pot is an example of the abundant ponds that, collectively, contribute crucially to species diversity. Despite extensive biological study, little has been reported about the physical framework that supports its ecological richness. This article elucidates the physical character of Priest Pot’s water column and thus that of similar water bodies. Vertical thermal microstructure profiles were recorded during summer 2003 and analyzed alongside concurrent meteorological data. During summer stratification, the thermal structure appeared to be dominated by surface heat fluxes. Surface wind stress, limited by sheltering vegetation, caused turbulent overturns once a surface mixed layer was present but appeared to contribute little to setting up the thermal structure. Variations in full-depth mean stratification occurred predominantly over seasonal and ∼5-day time scales, the passage of atmospheric pressure systems being posited as the cause of the latter. In the uppermost ∼0.5 m, where the stratification varied at subdaily time scales, turbulence was active (sensu Ivey and Imberger 1991) when this layer was mixed, with dissipation values ε ∼ 10⁻⁸ m² s⁻³ and vertical diffusivity K_Z = 10⁻⁴ — 10⁻⁶ m² s⁻¹. Where the water column was stratified, turbulence was strongly damped by both buoyancy and viscosity, and K_Z was an order of magnitude smaller. Vertical transport in the mixed layer occurred via many small overturns (Thorpe scale r.m.s. and maximum values were typically 0.02 m and 0.10 m, respectively), and seston were fully mixed through the water column.     

Larval recruitment of the blue mussel Mytilus edulis: The effect of flow and algae

The mussel Mytilus edulis settlement and distribution was studied on plastic panels with manipulated flow regime (faired, bluff, split and angled) with or without water soluble metabolites of the green alga Cladophora rupestris. The panels were exposed vertically on a device (hydrovane) that ensures their constant orientation in the current during the peak of larval settlement at 1 m depth. In order to investigate larval distribution on the panels, half of them were coated with a silicone vacuum grease that prevents larvae from de-attachment. This grease was not toxic and did not attract or repel larvae. Low densities of larvae on the un-greased plates compared to the greased ones suggested that some of larvae left the substratum. The blue mussel larvae initially settled in regions of reduced shear velocity and then redistribute to the regions of high shear velocity. The presence of the alga increased the density of blue mussel larvae and changed their distribution on the panels. Overall, our results demonstrated that larval recruitment of M. edulis is an active process affected both by boundary-layer hydrodynamics and algal waterborne compounds.     

Effect of larval ontogeny, turbulence and light on prey attack rate and swimming activity in herring larvae

The effects of ontogeny (larval size), light and turbulence on the attack rate and swimming activity (proportion of time swimming and duration of swimming bout) of herring larvae (15-28 mm TL) have been investigated. Emphasis was put on the experimental design in order to create a set-up where the turbulence intensity distribution could be accurately measured as well as controlled in the entire experimental tank.Both larval size (ontogeny) and light had a significant positive effect on prey attack rate. Likewise, an intermediate increase in turbulence had a positive effect on prey attack rate, but this effect was dependent of light intensity and larval size.At low light (1.5 μE m² s⁻¹) intermediate turbulence increased the prey attack rate significantly for larger larvae (26 and 28 mm), while at high light (18 μE m² s⁻¹) intermediate turbulence had only a significant positive effect on the attack rate of smaller larvae 20 and 23 mm.In general, our data show a dome-shaped response of turbulence on attack rate and a U-shaped response of turbulence on swimming activity.For herring larvae >20 mm, the maximum (attack rate) and minimum (swimming activity) response of turbulence were found at intermediate turbulence intensities (energy dissipation rates between 7∗10⁻⁸ and 1∗10⁻⁶ W/kg). The highest turbulence level tested (8∗10⁻⁶ W/kg) showed only negative effects, as attack rates where at the lowest and swimming activity at the highest.Swimming activity increased with larval size or light, and decreased at intermediate turbulence. Compared to turbulent intensities under natural conditions this implies that larger herring larvae at 10 m depth have to be exposed to wind speeds of more than 17 m/s before negative effects on attack rate and swimming activity occurs.     

Turbulence increases the risk of microcystin exposure in a eutrophic lake (Lake Taihu) during cyanobacterial bloom periods

Toxic cyanobacterial harmful algal blooms (CyanoHABs) have posed serious water use and public health threats because of the toxins they produce, such as the microcystins (MCs). The direct physical effects of turbulence on MCs, however, have not yet been addressed and is still poorly elucidated. In this study, a 6-day mesocosm experiment was carried out to evaluate the effects of wind wave turbulence on the competition of toxic Microcystis and MCs production in highly eutrophicated and turbulent Lake Taihu, China. Under turbulent conditions, MCs concentrations (both total and extracellular) significantly increased and reached a maximum level 3.4 times higher than in calm water. Specifically, short term (∼3 days) turbulence favored the growth of toxic Microcystis species, allowing for the accumulation of biomass which also triggered the increase in MCs toxicity. Moreover, intense turbulence raises the shear stress and could cause cell mechanical damage or cellular lysis resulting in cell breakage and leakage of intracellular materials including the toxins. The results indicate that short term (∼3 days) turbulence is beneficial for MCs production and release, which increase the potential exposure of aquatic organisms and humans. This study suggests that the importance of water turbulence in the competition of toxic Microcystis and MCs production, and provides new perspectives for control of toxin in CyanoHABs-infested lakes.     

Benthic shear stress gradient defines three mutually exclusive modes of non-biological internal nutrient loading in shallow lakes

Assessment of the importance of internal nutrient loading is essential for managing and restoring eutrophic shallow lakes. To date, studies of internal loads have tended to focus on one of two abiotic processes, either molecular diffusion or sediment/nutrient entrainment (resuspension). This study presents a new approach to determining the non-biological fluxes of nitrogen (N) and phosphorus (P) from the sediment to the water column of shallow lakes. Three mutually exclusive flux processes: (i) molecular diffusion, (ii) turbulent diffusion (eddy diffusivity) and (iii) wind-induced resuspension of N and P, were related to a gradient of benthic shear stress. A model presented here allowed the durations and magnitudes of different non-biological fluxes to be calculated over time, based on benthic shear stress. Two site-specific critical shear stress thresholds determined which of the three flux processes dominated for any benthic shear stress value. The model was calibrated for a shallow lake and the continuous flux of nutrient from the sediment to the overlying water generated by each process during that period was calculated, enabling the estimation of the relative importance of each of the three flux processes over a one-year period. Wind-induced resuspension dominated the internal nutrient flux, operating for 38% of the time and contributing 0.9 T P year⁻¹ and 10.2 T N year⁻¹ to the internal nutrient load. In contrast, molecular diffusion only contributed 0.01–0.02 T P year⁻¹ and 0.12–0.20 T N year⁻¹ to the water column, while turbulent diffusion provided up to 0.6 T P year⁻¹ and 6.2 T N year⁻¹. Our model suggests that turbulent diffusion is a neglected and potentially important process contributing to internal nutrient loading in shallow lakes, whereas molecular diffusion appears to be relatively unimportant in lakes that experience turbulence at the sediment–water interface. Handling editor: L. Naselli-Flores     

Environmental influences on the replenishment of lizardfish (family Synodontidae) in Caribbean Panama

Lizardfish (family Synodontidae) are little studied despite their potentially important predatory role in epibenthic coral reef communities. The present study documented the temporal and spatial larval supply patterns of five Caribbean lizardfish species together with environmental variables (solar radiation, rainfall, water temperature, onshore-offshore winds, alongshore winds and wind-induced turbulence) to examine: (1) whether species within the same family respond differently to their developmental environment and (2) if larval supply in year-round warm waters is influenced by climatic changes. To address these questions, late-stage larvae of Synodus foetens, Synodus intermedius, Synodus poeyi, Saurida suspicio and Saurida brasiliensis were collected in replicate light traps in three different reef habitats (back-reef, lagoon and exposed) in the San Blas Archipelago, Panama, over 18 consecutive lunar months. Although replenishment of lizardfish occurred year-round, the temporal and spatial supply patterns were species-specific: S. foetens, S. intermedius and S. poeyi were most abundant during the dry season while S. suspicio and S. brasiliensis were most prevalent during the wet season. When seasons were analysed separately, water temperature explained 39 and 26% of the variance in light trap catches of S. foetens and S. intermedius, respectively, in the dry season while wind-induced turbulence accounted for 25% of the variability in S. suspicio and S. brasiliensis catches during the wet season. These findings stress the importance of analysing larval supply in conjunction with environmental data at a high taxonomic resolution to better understand the mechanisms that drive replenishment in reef systems at low latitudes.     

Probabilistic settling in the Local Exchange Model of turbulent particle transport

The Local Exchange Model (LEM) is a stochastic diffusion model of particle transport in turbulent flowing water. It was developed mainly for application to particles of near-neutral buoyancy that are strongly influenced by turbulent eddies. Turbulence can rapidly transfer such particles to the bed, where settlement can then occur by, for example, sticking to biofilms (e.g., fine particulate organic matter, or FPOM) or attaching to the substrate behaviorally (e.g., benthic invertebrates). Previous papers on the LEM have addressed the problems of how long (time) and far (distance) a suspended particle will be transported before hitting the bed for the first time. These are the hitting-time and hitting-distance problems, respectively. Hitting distances predicted by the LEM for FPOM in natural streams tend to be much shorter than the distances at which most particles actually settle, suggesting that particles usually do not settle the first time they hit the bed. The present paper extends the LEM so it can address probabilistic settling, where a particle encountering the bed can either remain there for a positive length of time (i.e., settle) or immediately reflect back into the water column, each with positive probability. Previous results for the LEM are generalized by deducing a single set of equations governing the probability distribution and moments of a broad class of quantities that accumulate during particle trajectories terminated by hitting or settling on the bed (e.g., transport time, transport distance, cumulative energy expenditure during transport). Key properties of the settling-time and settling-distance distributions are studied numerically and compared with the observed FPOM settling-distance distribution for a natural stream. Some remaining limitations of the LEM and possible means of overcoming them are discussed.     

Numerical simulations of the pulsating flow of cerebrospinal fluid flow in the cervical spinal canal of a Chiari patient

The flow of cerebrospinal fluid (CSF) in a patient-specific model of the subarachnoid space in a Chiari I patient was investigated using numerical simulations. The pulsating CSF flow was modeled using a time-varying velocity pulse based on peak velocity measurements (diastole and systole) derived from a selection of patients with Chiari I malformation. The present study introduces the general definition of the Reynolds number to provide a measure of CSF flow instability to give an estimate of the possibility of turbulence occurring in CSF flow. This was motivated by the fact that the combination of pulsating flow and the geometric complexity of the spinal canal may result in local Reynolds numbers that are significantly higher than the commonly used global measure such that flow instabilities may develop into turbulent flow in these regions. The local Reynolds number was used in combination with derived statistics to characterize the flow. The results revealed the existence of both local unstable regions and local regions with velocity fluctuations similar in magnitude to what is observed in fully turbulent flows. The results also indicated that the fluctuations were not self-sustained turbulence, but rather flow instabilities that may develop into turbulence. The case considered was therefore believed to represent a CSF flow close to transition.     

Bloom formation potential in the harmful dinoflagellate Akashiwo sanguinea: Clues from movement behaviors and growth characteristics

We measured the growth rates and swimming behaviors of recently isolated strains of the dinoflagellate Akashiwo sanguinea to investigate to what degree growth and motility could contribute to the formation of in situ blooms. To quantify the effect of variation in in situ conditions on population growth rate, we applied two temperature treatments (10 °C and 20 °C) and measured growth in still conditions and on a shaker table, to emulate mild turbulence. To quantify the importance of intra-strain variability and trait variation in the species growth potential and vertical distribution, we included six strains isolated from a spatially and temporally extensive bloom on the US West Coast. Overall, as reported previously, A. sanguinea was observed to tolerate conditions amounting to a broad ecological niche with intra-specific variability further broadening tolerable conditions. In agreement with prior observations of slow growth rates of the species, average growth rates across all strains increased significantly from 0.12 d⁻¹ (±0.03) at 10 °C to 0.28 d⁻¹ (±0.13) at 20 °C in still conditions. Contrary to prior reports, mild turbulence had neutral or positive effects on most strains’ growth rates, with one strain only able to grow on the shaker table. Growth rates in mild turbulence were higher than in still conditions and increased from 0.15 d⁻¹ (±0.01) at 10 °C to 0.43 d⁻¹ (± 0.04) at 20 °C. There was significant intra-strain variation in growth rates (>50% coefficient of variation) and movement behaviors. All strains had both up and down swimming fractions, leading to predictions of vertically patchy distributions, rather than surface aggregations. Slow growth rates and dispersive swimming behaviors suggest in situ mortality must be low and tolerance of seasonally varying water temperatures lead to accumulation and persistence of cells over months and kilometers. Estimates of in situ loss rates are a critical but missing component of identifying the bloom formation mechanisms of this species.