A method for measuring depth-integrated community metabolism in experimental planktonic–benthic ecosystems

We developed incubation chambers of clear acrylic extending the full depth of the water column to measure in situ depth-integrated community metabolism in experimental planktonic–benthic ecosystems. A magnetic stirring apparatus at mid-depth in each chamber prevented stratification and maintained a vertical turnover time that roughly matched turnover in the experimental ecosystems. We used the chambers in light–dark experiments to partition the depth-integrated photosynthesis and respiration occurring in the experimental ecosystems among water column, wall periphyton, and benthic communities. Community metabolism measured with the chambers was positively related to the chlorophyll-a concentrations of the water column and wall periphyton communities. We also found close agreement between the summation of partitioned community metabolism measured in the chambers and an independent measure of total ecosystem metabolism in the experimental ecosystems. Incubations in chambers and in light–dark bottles revealed similar patterns of water column and wall periphyton community metabolism, but magnitudes differed. In addition to application in experimental ecosystems, this incubation chamber technique may provide an efficient and realistic measure of water column and benthic metabolism and nutrient uptake in lakes and estuaries.     

Mathematical models for estimating fluxes at the sediment-water interface in benthic chamber experiments

Experimental data collected by means of benthic chambers have been used to develop simple mathematical models in order to predict heavy metal and non-metal fluxes at the sediment-water interface. In particular, two mathematical models have been applied to Ba, Co, Fe, Pb, Mn and Cu.Both models estimate benthic fluxes by means of Fick's law but they differ in the modelling of partition between liquid and solid phases. The first model, which is appropriate for Ba and Co, assumes that sorption is the most important process in determining partition. The second model, which gives discrete results for Fe and Pb, assumes that precipitation is the most important process determining the time variation of concentrations. Neither model is suitable to explain experimental data for Mn and Cu.This paper demonstrates the benefit of the use of mathematical models in the interpretation of benthic chambers experiments and in the identification of the relevant processes occurring at the sediment-water interface.     

Aquatic Eddy Correlation: Quantifying the Artificial Flux Caused by Stirring-Sensitive O2 Sensors

In the last decade, the aquatic eddy correlation (EC) technique has proven to be a powerful approach for non-invasive measurements of oxygen fluxes across the sediment water interface. Fundamental to the EC approach is the correlation of turbulent velocity and oxygen concentration fluctuations measured with high frequencies in the same sampling volume. Oxygen concentrations are commonly measured with fast responding electrochemical microsensors. However, due to their own oxygen consumption, electrochemical microsensors are sensitive to changes of the diffusive boundary layer surrounding the probe and thus to changes in the ambient flow velocity. The so-called stirring sensitivity of microsensors constitutes an inherent correlation of flow velocity and oxygen sensing and thus an artificial flux which can confound the benthic flux determination. To assess the artificial flux we measured the correlation between the turbulent flow velocity and the signal of oxygen microsensors in a sealed annular flume without any oxygen sinks and sources. Experiments revealed significant correlations, even for sensors designed to have low stirring sensitivities of ~0.7%. The artificial fluxes depended on ambient flow conditions and, counter intuitively, increased at higher velocities because of the nonlinear contribution of turbulent velocity fluctuations. The measured artificial fluxes ranged from 2 - 70 mmol m^-2 d^-1 for weak and very strong turbulent flow, respectively. Further, the stirring sensitivity depended on the sensor orientation towards the flow. For a sensor orientation typically used in field studies, the artificial flux could be predicted using a simplified mathematical model. Optical microsensors (optodes) that should not exhibit a stirring sensitivity were tested in parallel and did not show any significant correlation between O₂ signals and turbulent flow. In conclusion, EC data obtained with electrochemical sensors can be affected by artificial flux and we recommend using optical microsensors in future EC-studies.     

A charcoal cartridge for the removal of anionic detergent and electrophoresis stains

Instructions are given for the construction of a charcoal-containing cartridge that allows the rapid recirculation through charcoal of any fluid in which the cartridge is submerged; recirculatory flow is achieved by magnetic stirring of the fluid by a stirring bar placed under the cartridge. The device is assembled from nylon mesh and conical sections cut from polypropylene beakers. The device can be used to accelerate the destaining of electrophoresis gels and to remove SDS (sodium lauryl sulfate) from SDS gels. The removal of SDS prior to staining is essential for the staining of SDS gels with Coomassie Brilliant Blue G-250.     

A simple filtration device for separating nonadherent microorganisms from mammalian cells in in vitro studies on microbial adherence

In adherence studies, the removal of nonadherent microorganisms is essential for the valid enumeration of microorganisms that adhere to host cells. Although filtration devices are available commercially for the removal of nonadherent microorganisms, these are expensive and not reusable. In this article, we describe a simple, inexpensive, and reusable filtration device composed of two chambers of nylon, a nylon membrane of desired pore size, a rubber washer, and supporting stainless steel mesh. The device was effective in in vitro adherence assays for removing nonadherent endospores of Rhinosporidium seeberi from human buccal epithelial cells, providing valid counts of adherent microorganisms.     

A pedal corer for the quantitative sampling of sediments and benthic organisms in submerged areas accessible on foot

This paper presents a new device to collect quantitative samples of sediment and benthic organisms. The device is specially designed for sampling with the advantages of box-corer or Eckman dredges in submerged areas that are accessible on foot. The pedal corer is a simple, lightweight, user-friendly device that does not disturb the sediment structure and provides easy access to the sample contained inside the core. With this device, sampling in shallow water zones that are constantly submerged is made easy and sampling time is extended in intertidal zones. Handling editor: J. Saros     

Dams and Flow in the Cotter River, Australia: Effects on Instream Trophic Structure and Benthic Metabolism

This study assessed benthic macroinvertebrates and periphyton and its responses to managed river-flows, in riffles downstream of three dams on the Cotter River, Australian Capital Territory. Benthic macroinvertebrates and periphyton were also assessed in adjacent tributaries of the river, as well as in a nearby unregulated river and its tributaries. Food sources of four macroinvertebrate taxa (Leptophlebiidae, Elmidae, Glossosomatidae and Orthocladiinae) were determined by stable isotope analysis of the invertebrates and their potential food, in conjunction with examination of the gut contents of individual invertebrates. Components of benthic periphyton were the main food source for the selected taxa. Orthocladiinae consumed primarily amorphous detritus, while Elmidae, Glossosomatidae and Leptophlebiidae consumed diatoms. Enclosed benthic chambers were used to measure the response of benthic metabolism to monthly flow spikes released from one of the dams. The balance of benthic metabolism as measured by the Production/Respiration ratio (P/R) showed a shift towards production after the release of flow spikes. At sites downstream of the dams, there was more periphyton chlorophyll-a in the form of filamentous green algae than at sites in the unregulated river and the tributaries, and macroinvertebrate taxa using periphyton as a food resource were missing or reduced in abundance relative to sites without dams. However, the site downstream of the dam with environmental flow releases had more macroinvertebrate taxa and less periphyton cholorophyll-a content than sites downstream of dams without managed environmental flows, suggesting that a more suitable food supply resulting from environmental flow releases shifted macroinvertebrate communities towards those of unregulated streams.     

Microchambers for inverted microscopes

The designs for two microchambers, non-flow and flow, for inverted microscopes are described. These chambers are intended to study dynamic processes in living cells. The chambers described are made completely of a glass. An advantage of the flow chamber is the possibility of making simple changes in its internal space. It is dismantled easily to allow careful cleaning. This is especially important when working with toxic substances.     

Microchambers for inverted microscopes

The designs for two microchambers, non-flow and flow, for inverted microscopes are described. These chambers are intended to study dynamic processes in living cells. The chambers described are made completely of a glass. An advantage of the flow chamber is the possibility of making simple changes in its internal space. It is dismantled easily to allow careful cleaning. This is especially important when working with toxic substances.     

A device for sample pretreatment during flow cytometry]

A device for the preliminary treatment of samples immediately prior to flow cytofluorimetric analysis is described. The device is intended for several procedures: (a) mixing of batched sample volumes with the reagent and efficient stirring of the mixture; (b) disintegration of cell aggregates; and (c) disruption of cell membranes to release the cell contents (chromosomes, micronuclei, nuclei etc.). The pretreatment is useful for studying the kinetic parameters of fast cellular processes in the flow, a more correct analysis of the cell cycle and the study of karyotypes of single mitotic cells. The device was called a magnetic microstirrer.     

Tissue fluid pressure and flow during pneumatic compression in lymphedema of lower limbs

Background: Physiotherapy of edema in cases with obstructed main lymphatics of lower limbs requires knowledge of how high external pressures should be applied manually or set in compression devices in order to generate tissue pressures high enough to move tissue fluid to nonswollen regions and to measure its flow rate. Methods: We measured tissue fluid pressure and flow in subcutaneous tissue of lymphedematous limbs stages II to IV at rest and during pneumatic compression under various pressures and inflation timing. An 8-chamber sequential compression device inflated to pressures 50-120?mmHg, for 50 sec each chamber, with no distal deflation, was used. Pressures were measured using a wick-in-needle and electronic manometer. Fluid flow was calculated from continuously recorded changes in limb circumference using strain gauge plethysmography. Results: Before massage, in all stages of lymphedema, stagnant tissue fluid pressures in subcutaneous tissue ranged between -1 and +10 mmHg and did not differ from those measured in normal subjects. Pressures generated in tissue fluid by pneumatic compression reached 40-100 mmHg and were lower than those in inflated chambers. High pressure gradient through the skin was caused by its rigidity (fibrosis) and dissipation of applied compression force to proximal noncompressed limb regions. The calculated volumes of displaced tissue fluid ranged from 10 to 30 ml per compression cycle, to reach in some cases 100 ml in the groin region. Conclusions: Tissue fluid pressures generated by a pneumatic device were found lower than in the compression chambers. The obtained results point to the necessity of applying high pressures and longer compression times to generate effective tissue fluid pressures and to provide enough time for moving the stagnant fluid.     

Resuspension studies in cylindrical microcosms: Effects of stirring velocity on the dynamics of redox sensitive elements in a coastal sediment

Effects of resuspension on the release of dissolved, redox sensitive elements (Fe, Mn) was studied in cylindrical microcosms. Effects from changing water stirring velocity in sediment pools were evaluated through measurements of pore water profiles of dissolved Mn, Fe and redox potential. Mn was a good natural marker to follow such effects. At current velocities below the threshold velocity for resuspension (37 cm s-1), Mn release rates to overlying water were 100 times higher compared to steady-state values. Pulse increases in Mn concentration were the result of convective currents inside flow chambers. These results were strongly supported by measurements of Eh profiles in the sediment pore water. Furthermore, impacts from increasing stirring velocity were found down to 1.9 cm depth below the resuspended layer of sediment.     

Osmotic flow caused by polyelectrolytes

Osmotic flow of water caused by high concentrations of anionic polyelectrolytes across semipermeable membranes, permeable only to solvent and simple electrolyte, has been measured in a newly designed flow cell. The flow cell features small solution and solvent compartments and an efficient stirring mechanism. We have demonstrated that, while the osmotic pressure of the anionic polyelectrolytes is determined primarily by micro-counterions, the osmotic flow is determined by solution-dependent properties as embodied in the hydrodynamic frictional coefficient which is determined by the polymer backbone segment of the polyelectrolyte. The variation of the osmotic permeability coefficient, L(p)(o), with concentration and osmotic pressure closely correlated with the concentration dependence of this frictional coefficient. These studies confirm previous work that the kinetics of osmotic flow across a membrane impermeable to the osmotically active solute is primarily determined by the diffusive mobility of the solute.     

A microbead array chemical sensor using capillary-based sample introduction: toward the development of an "electronic tongue"

The development of a micromachined fluidic structure for the introduction of liquid samples into a chip-based sensor array composed of individually addressable polymeric microbeads is presented. The micromachined structure consists of micromachined storage cavities combined with a covering glass layer that confines the microbeads and fluidic channels. In our sensor array transduction occurs via optical (colorimetric and fluorescence) changes to receptors and indicator molecules that are covalently attached to termination sites on the polymeric microbeads. Spectral data are acquired for each of the individual microbeads using a charged-coupled device (CCD) allowing for the near-real-time analysis of liquid sample. Hence the micromachined fluidic structure must allow for both optical access to the microbeads and fluid flow through the micromachined cavities that serve as the microreactors/analysis chambers. One of the key parts of the structure is a passive fluid introduction system driven only by capillary force. This simple means of fluid introduction realizes a compact device. The capillary flow on the inlet channel has been studied, and the responses of the microbeads (alizarin complexone) to a liquid sample have been characterized. The test results show that this system is useful in a micro-total-analysis-system (mu-TAS) and biomedical applications.     

An automated system for detection and analysis of locomotor behavior in crustaceans

An efficient and simple system is presented for the analysis of crustacean locomotor behavior. The system is composed by six dual-compartment actographic chambers with photocoupling circuits for movement detection, and a device for acquisition and analysis of data. Such device is made by a digital interface which feeds into a microcomputer with disc unit and printer. Information is processed in real time during the experiment, with a simultaneous printout and storage in a floppy disc.     

Factors influencing the spatio-temporal distribution of benthic Microcystis aeruginosa colonies (Cyanobacteria) in the hypertrophic Grangent Reservoir (Loire, France)

The spatio-temporal distribution of benthic colonies of Microcystis aeruginosa in Grangent Reservoir (France) in 2000 was not homogeneous and appeared to be controlled by many external factors: lake depth, station morphometry, substratum and hydraulic regime (lacustrine or fluvial). A most important concentration of benthic colonies was found at deep sites with fine sediment or at sites where the sediment was rich in organic matter. In spite of a stable water level and a minimum flow during summer, the number of benthic colonies showed great variation in the lacustrine downstream part of the reservoir. These variations may be explained by the dynamics of planktonic cyanobacteria.     

A portable flow chamber for in situ determination of benthic metabolism

1. Many stream ecologists are interested in determining the metabolic rates of benthic organisms, particularly those of production and respiration. It is often necessary to make these measurements on fresh material in the field at remote sites. Recirculating chambers are commonly used for this purpose.
2. A broad variety of recirculating chambers are described in the literature, but each design has inherent limitations. The most common are inability to control flow in the chamber and match it with external flow rates, and a lack of the power required to do this for extended periods. Alteration of spectral irradiance, temperature rise and elevated internal chamber pressures are additional limitations that have received little attention.
3. We have designed and constructed a flow chamber that eliminates some of these problems. The chamber utilizes a DC motor-driven propeller as an efficient recirculator (axial impeller), minimizing power requirements and it is constructed of UVB transparent acrylic to allow a full spectral complement of solar irradiance in the interior. Modular components allow the chamber to be taken apart quickly for cleaning and replacement of parts, making it more functional than some previous designs.
4. The axial impeller chamber was compared to a similar sized conventional chamber that had a small diameter return line and a high capacity centrifugal pump. The axial impeller chamber had less of a temperature rise during field incubations, lower power consumption and less internal pressure in the return line when producing equivalent water velocities.
5. The reported axial impeller design had relatively homogeneous velocity across the working section relative to other chambers and was capable of water velocities in excess of 1 m s^–1.     

Effects of eutrophication on the interaction between algae and grazers in an Andean stream

Nutrient excess is a common disturbance that affects biological interactions in river ecosystems. The response of nutrient supply on primary producers and Tricorythodes sp., a common mayfly grazer, was determined in experimental chambers set in a tropical, high Andean stream. Chambers in an experimentally fertilized reach developed higher amount of both benthic and detached chlorophyll than chambers in an upstream control reach. Fertilization produced a slight increase in grazer biomass, and reduced algal biomass compared to grazer-free chambers. These results show that nutrient excess spread bottom-up effects through the food web, and that relevant top-down effects could also be detected. Eutrophication may produce relevant changes in the food web of tropical high-mountain streams.     

An inexpensive device for sample stirring in VSU Zeiss spectrophotometers. Its use for kinetic measurements and active site titrations with immobilized enzymes.

The construction of a simple and effective sample stirring device for commercial spectrophotometers and its use for continuous kinetic measurements and active site titrations with immobilized enzymes is described. Sepharose-bound leucine aminopeptidase and trypsin were selected as model enzymes to test the performance of the magnetic stirring equipment. Kinetic parameters of insolubilized leucine aminopeptidase using L-leucine p-nitroanilide as substrate and the catalytic site concentration of matris-bound trypsin using p-nitrophenyl p'-guanidinobenzoate as active site titrant could be evaluated without significant interference from the turbidity of the stirred Sepharose suspension. The problem of grinding of the support material could be overcome. Both unbound native and carrier-fixed enzyme may be reacted under identical conditions with similar convenience and sensitivity.     

Morphological abnormalities in Late Cretaceous and early Paleocene foraminifer tests (northern Patagonia, Argentina)

Morphological abnormalities are common in Late Cretaceous and early Paleocene foraminifer tests at two localities in northern Patagonia, Argentina. Protelphidium sp. in the Auca Mahuevo section (late Campanian–early Maastrichtian) exhibit abnormal size or shape of the later chambers, with the last chamber commonly larger than normal or inflated and variably extending onto one of the lateral sides of the test; modification of the coiling plane; protuberances near the proloculus or on one or more chambers; a double last chamber, and complex forms. Protelphidium hofkeri Haynes in the Cerro Azul section (Danian) exhibit abnormal size or shape of one or more chambers, producing peripheral irregularities. In addition, there are rare multiple tests in planktic species from the Cerro Azul section, probably teratological specimens. The sedimentology of the sections and the character of the accompanying faunas indicate that the abnormalities in the two benthic foraminiferal taxa were most probably caused by hypersalinity and/or fluctuations in salinity. The fossil occurrence of assemblages with abundant deformed specimens suggests that investigators should carefully look at many aspects of the environment before concluding that anthropogenic pollution is the only cause of deformations of living benthic foraminifera.