Evaluation of the attachment strength of individuals of Asterina gibbosa (Asteroidea,Echinodermata) during the perimetamorphic period

Abstract

A turbulent channel flow apparatus was used to determine the adhesion strength of the three perimetamorphic stages of the asteroid Asterina gibbosa, i.e. the brachiolaria larvae, the metamorphic individuals and the juveniles. The mean critical wall shear stresses (wall shear stress required to dislodge 50% of the attached individuals) necessary to detach larvae attached by the brachiolar arms (1.2 Pa) and juveniles attached by the tube feet (7.1 Pa) were one order of magnitude lower than the stress required to dislodge metamorphic individuals attached by the adhesive disc (41 Pa). This variability in adhesion strength reflects differences in the functioning of the adhesive organs for these different life stages of sea stars. Brachiolar arms and tube feet function as temporary adhesion organs, allowing repetitive cycles of attachment to and detachment from the substratum, whereas the adhesive disc is used only once, at the onset of metamorphosis, and is responsible for the strong attachment of the metamorphic individual, which can be described as permanent adhesion. The results confirm that the turbulent water channel apparatus is a powerful tool to investigate the adhesion mechanisms of minute organisms.     

Removal kinetics of Bacillus cereus spores from a stainless steel surface exposed to constant shear stress 1 ‐ experimental system

The efficiency of cleaning in place procedures in dairy industries can be greatly affected by the presence of spore‐forming bacteria, which are able to adhere strongly to surfaces and to survive disinfection procedures. Microbial adhesion has been extensively studied, but very few studies have yet reported on the hydrodynamic removal of microorganisms, due to the lack of simple, routinely performable techniques. In this paper, a methodology using a coaxial cylinder double gap viscometer is described, to study the removal kinetics of Bacillus cereus spores from a stainless steel support under hydrodynamic conditions. This method was shown to be reproducible, sensitive and easy to perform, and allowed spore hydrodynamic removal kinetics to be studied as a function of both adhesion and detachment conditions. A high ionic strength attachment medium was shown to enhance adhesion forces, provided it did not contain macromolecules. An increase in shear stress was found to be favorable to spore detachment (4 to 5 times more spores were detached at 28 Pa than at 2 Pa), but removal kinetics were not found to be significantly different for 2 and 15 Pa. Thus, the effect of shear stress on spore removal kinetics may not be linear.     

Biophysical Response to Pulsed Laser Microbeam‐Induced Cell Lysis and Molecular Delivery

Cell lysis and molecular delivery in confluent monolayers of PtK₂ cells are achieved by the delivery of 6 ns, λ = 532 nm laser pulses via a 40×, 0.8 NA microscope objective. With increasing distance from the point of laser focus we find regions of (a) immediate cell lysis; (b) necrotic cells that detach during the fluorescence assays; (c) permeabilized cells sufficient to facilitate the uptake of small (3 kDa) FITC‐conjugated Dextran molecules in viable cells; and (d) unaffected, viable cells. The spatial extent of cell lysis, cell detachment, and molecular delivery increased with laser pulse energy. Hydrodynamic analysis from time‐resolved imaging studies reveal that the maximum wall shear stress associated with the pulsed laser microbeam‐induced cavitation bubble expansion governs the location and spatial extent of each of these regions independent of laser pulse energy. Specifically, cells exposed to maximum wall shear stresses τ_{w, max} > 190 ± 20 kPa are immediately lysed while cells exposed to τ_{w, max} > 18 ± 2 kPa are necrotic and subsequently detach. Cells exposed to τ_{w, max} in the range 8–18 kPa are viable and successfully optoporated with 3 kDa Dextran molecules. Cells exposed to τ_{w, max} < 8 ± 1 kPa remain viable without molecular delivery. These findings provide the first direct correlation between pulsed laser microbeam‐induced shear stresses and subsequent cellular outcome. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Quantifying Cell Adhesion through Impingement of a Controlled Microjet

The impingement of a submerged, liquid jet onto a cell-covered surface allows assessing cell attachment on surfaces in a straightforward and quantitative manner and in real time, yielding valuable information on cell adhesion. However, this approach is insufficiently characterized for reliable and routine use. In this work, we both model and measure the shear stress exerted by the jet on the impingement surface in the micrometer-domain, and subsequently correlate this to jet-induced cell detachment. The measured and numerically calculated shear stress data are in good agreement with each other, and with previously published values. Real-time monitoring of the cell detachment reveals the creation of a circular cell-free area upon jet impingement, with two successive detachment regimes: 1), a dynamic regime, during which the cell-free area grows as a function of both the maximum shear stress exerted by the jet and the jet diameter; followed by 2), a stationary regime, with no further evolution of the cell-free area. For the latter regime, which is relevant for cell adhesion strength assessment, a relationship between the jet Reynolds number, the cell-free area, and the cell adhesion strength is proposed. To illustrate the capability of the technique, the adhesion strength of HeLa cervical cancer cells is determined ((34 ± 14) N/m²). Real-time visualization of cell detachment in the dynamic regime shows that cells detach either cell-by-cell or by collectively (for which intact parts of the monolayer detach as cell sheets). This process is dictated by the cell monolayer density, with a typical threshold of (1.8 ± 0.2) × 10⁹ cells/m², above which the collective behavior is mostly observed. The jet impingement method presents great promises for the field of tissue engineering, as the influence of both the shear stress and the surface characteristics on cell adhesion can be systematically studied.     

The influence of surface wettability on the adhesion strength of settled spores of the green alga enteromorpha and the diatom amphora

In this paper we report on the effect of surface wettabilityon surface selection and adhesion properties of settled (adhered)spores of the biofouling marine alga Enteromorpha and cellsof the diatom Amphora, through the use of patterned self-assembledmonolayers (SAMs). The SAMs were formed from alkanethiols terminatedwith methyl (CH₃) or hydroxyl (OH) groups, or mixtures of thetwo, creating a discontinuous gradient of wettability as measuredby advancing water contact angle. In the case of Enteromorpha,primary adhesion, as measured by the transition from a motilespore to a settled, sessile organism, is strongly promoted bythe hydrophobic surfaces. On the other hand, adhesion strengthof the settled spores, as measured by resistance to detachmentin a turbulent flow cell, is greatest on a hydrophilic surface.In the case of Amphora, there is little influence of surfacewettability on the primary adhesion of this organism, but motilityis inhibited at contact angles     

Attachment strength of an adhesive nuisance mussel,limnoperna fortunei,against water flow

The attachment strength of the freshwater mussel Limnoperna fortunei against water flow was studied. Newton's expression successfully described the hydrodynamic drag force acting on the mussel with a drag coefficient value of 1.03. The drag‐resistant force (defined as hydrodynamic drag force at mussel detachment) was smaller than the detachment force measured using a tensile load test. A fairly good correlation was obtained between the drag‐resistant force and the number of secreted threads. The drag‐resistant force divided by the number of threads increased with shell size, suggesting that byssal thread strength increased with mussel growth. For the mussel specimens obtained from a water transmission pipe, thread width increased with shell size. However, thread width was not dependent on current velocity. There was no correlation between the number of secreted threads and shell length, which indicated that the number of secreted threads did not change with mussel size. Therefore, the water velocity needed to detach mussels increases with shell size of the mussel when the number of secreted threads is constant. The increases in the water velocity to detach mussels with larger shells suggests that the mussel becomes more resistant to water flow as it grows. It is estimated that a flow velocity of around lms^‐1 is critical for attachment/detachment of a juvenile mussel with a shell length of a few millimeters and one hundred byssal threads.     

Ship hull in-water cleaning and its effects on fouling-control coatings

Abstract

Today, ship hull fouling is managed through fouling-control coatings, complemented with in-water cleaning. During cleaning, coating damage and wear must be avoided, for maximum coating lifetime and reduced antifoulant release. When possible, cleaning should target early stages of fouling, using minimal forces. However, such forces, and their effects on coatings, have not yet been fully quantified. In this one-year study, minimal cleaning forces were determined using a newly-designed immersed waterjet. The results show that bi-monthly/monthly cleaning, with maximum wall shear stress up to ～1.3?kPa and jet stagnation pressure ～0.17?MPa, did not appear to cause damage or wear on either the biocidal antifouling (AF) or the biocide-free foul-release (FR) coatings. The AF coating required bi-monthly cleanings to keep fouling to incipient slime (time-averaged results), while the FR coating had a similar fouling level even without cleaning. The reported forces may be used in matching cleaning parameters to the adhesion strength of the early stages of fouling.     

Effects of Combined Shear and Thermal Forces on Destruction of Microbacterium lacticum

A twin-screw extruder and a rotational rheometer were used to generate shear forces in concentrated gelatin inoculated with a heat-resistant isolate of a vegetative bacterial species, Microbacterium lacticum. Shear forces in the extruder were mainly controlled by varying the water feed rate. The water content of the extrudates changed between 19 and 45% (wet weight basis). Higher shear forces generated at low water contents and the calculated die wall shear stress correlated strongly with bacterial destruction. No surviving microorganisms could be detected at the highest wall shear stress of 409 kPa, giving log reduction of 5.3 (minimum detection level, 2 × 10⁴ CFU/sample). The mean residence time of the microorganism in the extruder was 49 to 58 s, and the maximum temperature measured in the end of the die was 73°C. The D_75°C of the microorganism in gelatin at 65% water content was 20 min. It is concluded that the physical forces generated in the reverse screw element and the extruder die rather than heat played a major part in cell destruction. In a rotational rheometer, after shearing of a mix of microorganisms with gelatin at 65% (wt/wt) moisture content for 4 min at a shear stress of 2.8 kPa and a temperature of 75°C, the number of surviving microorganisms in the sheared sample was 5.2 × 10⁶ CFU/g of sample compared with 1.4 × 10⁸ CFU/g of sample in the nonsheared control. The relative effectiveness of physical forces in the killing of bacteria and destruction of starch granules is discussed.     

A numerical analysis of forces exerted by laminar flow on spreading cells in a parallel plate flow chamber assay

Exposure of spreading anchorage-dependent cells to laminar flow is a common technique to measure the strength of cell adhesion. Since cells protrude into the flow stream, the force exerted by the fluid on the cells is a function of cell shape. To assess the relationship between cell shape and the hydrodynamic force on adherent cells, we obtained numerical solutions of the velocity and stress fields around bovine aortic endothelial cells during various stages of spreading and calculated the force required to detach the cells. Morphometric parameters were obtained from light and scanning electron microscopy measurements. Cells were assumed to have a constant volume, but the surface area increased during spreading until the membrane was stretched taut. Two-dimensional models of steady flow were generated using the software packages ANSYS (mesh generation) and FIDAP (problem solution). The validity of the numerical results was tested by comparison with published results for a semicircle in contact with the surface. The drag force and torque were greatest for round cells making initial contact with the surface. During spreading, the drag force and torque declined by factors of 2 and 20, respectively. The calculated forces and moments were used in adhesion models to predict the wall shear stress at which the cells detached. Based upon published values for the bond force and receptor number, round cells should detach at shear stresses between 2.5 and 6 dyn/cm(2), whereas substantially higher stresses are needed to detach spreading and fully spread cells. Results from the simulations indicate that (1) the drag force varies little with cell shape whereas the torque is very sensitive to cell shape, and (2) the increase in the strength of adhesion during spreading is due to increased contact area and receptor densities within the contact area. (c) 1993 John Wiley & Sons, Inc.     

Sporulation ofEnteromorpha spp. (Chlorophyta) and overwintering of spores in sediments of the Wadden Sea,Island Sylt,North Sea

For the last two decades dense mats of species of the filamentous green algaeEnteromorpha spp. have regulary occurred on tidal flats of Köningshafen Bay (island Sylt, North Sea, FRG). In calm areas overwintering of adult plants or plant fragments is a common process to guarantee the mass development during the next season. In contrast, the distribution ofEnteromorpha on exposed sandy tidal flats depends on recruitment by juvenile stages. In 1993Enteromorpha spore settlement was recorded regularly in the field. Polyethylene dishes were placed in the field and left for a period of seven days and lateron cultivated in the laboratory to checkEnteromorpha germling development. During summer 1993 — at a minimum distance of 200 m to the nearest adultEnteromorpha populations — a total of at least 82×10⁶ spores m^–2 settled. During winter the number of spores attached to the collecting dishes was close to zero and the adjacent sand flats were free of any visibleEnteromorpha plants. In further experiments it was shown that the development ofEnteromorpha juveniles in the next spring depended on the overwintering capacity of spores. More than 2×10⁶ spores m^–2 attached to large sand grains and other substrata (e.g. Hydrobia ulvae) survived the winter. In a laboratory experiment several species ofEnteromorpha were able to survive in total darkness for at least 10 months.     

Finite element sub-modeling analyses of damage to enamel at the incisor enamel/adhesive interface upon de-bonding for different orthodontic bracket bases

This study investigates the micro-mechanical behavior associated with enamel damage at an enamel/adhesive interface for different bracket bases subjected to various detachment forces using 3-D finite element (FE) sub-modeling analysis. Two FE macro-models using triangular and square bracket bases subjected to shear, tensile and torsional de-bonding forces were established using μCT images. Six enamel/adhesive interface sub-models with micro- resin tag morphology and enamel rod arrangement were constructed at the corresponding stress concentrations in macro-model results. The boundary conditions for the sub-models were determined from the macro-model results and applied in sub-modeling analysis. The enamel and resin cement stress concentrations for triangular and square bases were observed at the adhesive bottom towards the occlusal surface under shear force and at the mesial and distal side planes under tensile force. The corresponding areas under torsional force were at the three corners of the adhesive for the triangular base and at the adhesive bottom toward/off the occlusal surface for the square base. In the sub-model analysis, the concentration regions were at the resin tag base and in the region around the etched holes in the enamel. These were perfectly consistent with morphological observations in a parallel in vitro bracket detachment experiment. The critical de-bonding forces damaging the enamel for the square base were lower than those of the triangular base for all detached forces. This study establishes that FE sub-modeling can be used to simulate the stress pattern at the micro-scale enamel/adhesive interface, suggesting that a square base bracket might be better than a triangular bracket. A de-bonding shear force can detach a bracket more easily than any other force with a lower risk of enamel loss.     

Control of macroalgal blooms at early developmental stages: Pilayella littoralis versus Enteromorpha spp.

Although blooms of opportunistic fast-growing macroalgae now occur frequently in coastal ecosystems affected by eutrophication, their initiation and control is little understood. Most previous studies have focused on the ecophysiology of adult algae only. We show that spores and/or germlings may represent critical stages in the life cycles and mass-developments of co-occurring opportunistic macroalgae in the Baltic (Pilayella littoralis and Enteromorpha spp.). We investigated the overwintering of spores, timing of germination, subsequent growth, and grazing on spores and germlings, in order to explain the initiation of mass blooms and species dominance patterns. In the field, Enteromorpha spp. showed 10- to 50-fold higher abundances of overwintering microscopic forms (up to 330 individuals cm⁻²) than P. littoralis. Moreover, we found continuous production of spores (up to 1.2 million settling spores m⁻² h⁻¹) from April to October in Enteromorpha spp., while there was evidence of only a short reproductive period in Pilayella. However, in spring, germlings and adults of P. littoralis appeared earlier in the field and reached a 10-fold higher biomass than Enteromorpha spp. In factorial laboratory experiments including temperature and light, there were clear differences in timing of germination. P. littoralis germinated at 5°C whereas Enteromorpha spp. required temperatures of 10–15°C for germination. In contrast, we detected only minor differences in growth response among adults of P. littoralis and Enteromorpha spp. Germination, not growth of adults, appeared to be the ecophysiological bottleneck for initiating mass spring development. Following the spring Pilayella bloom, Enteromorpha germlings occurred massively in the field (April–September), but rarely developed into adults. In laboratory feeding experiments we tested whether crustacean mesograzers common in summer may control development of Enteromorpha germlings. Both germination of settled spores and growth of germlings were reduced by 93–99% in the presence of grazers (Idotea chelipes and Gammarus locusta). Thus in addition to ecophysiological constraints, grazers, if present, may play a decisive role in the early life stages of macroalgal mass developments. These results mirror patterns of overwintering of seeds, germination control, seed and seedling predation in terrestrial plant communities. Received: 7 March 1998 / Accepted: 18 November 1998     

Evaluation of the attachment strength of individuals of Asterina gibbosa (Asteroidea, Echinodermata) during the perimetamorphic period

A turbulent channel flow apparatus was used to determine the adhesion strength of the three perimetamorphic stages of the asteroid Asterina gibbosa, i.e. the brachiolaria larvae, the metamorphic individuals and the juveniles. The mean critical wall shear stresses (wall shear stress required to dislodge 50% of the attached individuals) necessary to detach larvae attached by the brachiolar arms (1.2 Pa) and juveniles attached by the tube feet (7.1 Pa) were one order of magnitude lower than the stress required to dislodge metamorphic individuals attached by the adhesive disc (41 Pa). This variability in adhesion strength reflects differences in the functioning of the adhesive organs for these different life stages of sea stars. Brachiolar arms and tube feet function as temporary adhesion organs, allowing repetitive cycles of attachment to and detachment from the substratum, whereas the adhesive disc is used only once, at the onset of metamorphosis, and is responsible for the strong attachment of the metamorphic individual, which can be described as permanent adhesion. The results confirm that the turbulent water channel apparatus is a powerful tool to investigate the adhesion mechanisms of minute organisms.     

Adhesion of Azospirillum brasilense to polystyrene: Influence of ionic strength through protein adsorption

The influence of ionic strength on the adhesion of Azospirillum brasilense to polystyrene has been examined by comparing water and phosphate buffer saline (PBS) as suspending media. Polystyrene supports analysed by X‐ray photoelectron spectroscopy (XPS) after adhesion in PBS for 2 h or 24 h and detachment of adhering cells showed a higher protein surface concentration, reflected by the N/C atomic concentration ratio, compared to supports analysed after adhesion in water. It was shown that PBS both favours protein release by the cells into the solution and enhances the tendency of proteins to adsorb at the support surface.

After 2 h contact time, the increase in the concentration of adsorbed proteins in PBS was related to an increase in adhesion density. However, the observation that the adhesion density after 24 h was lower in PBS than in water indicated that the amount of proteins adsorbed at the support surface controls cell adhesion in a complex way. In PBS, a thick layer of proteinaceous material retaining the bacterial cells is formed; this leads to underestimation of the density of adhering cells as well as to a heterogeneous adhesion pattern and to a relatively low adhesion density due to detachment of pellicles upon rinsing.

The ionic strength thus influences bacterial adhesion in a more subtle way than simply through double layer interactions between the cells and the support.     

Effects of combined shear and thermal forces on destruction of Microbacterium lacticum.

A twin-screw extruder and a rotational rheometer were used to generate shear forces in concentrated gelatin inoculated with a heat-resistant isolate of a vegetative bacterial species, Microbacterium lacticum. Shear forces in the extruder were mainly controlled by varying the water feed rate. The water content of the extrudates changed between 19 and 45% (wet weight basis). Higher shear forces generated at low water contents and the calculated die wall shear stress correlated strongly with bacterial destruction. No surviving microorganisms could be detected at the highest wall shear stress of 409 kPa, giving log reduction of 5.3 (minimum detection level, 2 x 10(4) CFU/sample). The mean residence time of the microorganism in the extruder was 49 to 58 s, and the maximum temperature measured in the end of the die was 73 degrees C. The D(75 degrees C) of the microorganism in gelatin at 65% water content was 20 min. It is concluded that the physical forces generated in the reverse screw element and the extruder die rather than heat played a major part in cell destruction. In a rotational rheometer, after shearing of a mix of microorganisms with gelatin at 65% (wt/wt) moisture content for 4 min at a shear stress of 2.8 kPa and a temperature of 75 degrees C, the number of surviving microorganisms in the sheared sample was 5.2 x 10(6) CFU/g of sample compared with 1.4 x 10(8) CFU/g of sample in the nonsheared control. The relative effectiveness of physical forces in the killing of bacteria and destruction of starch granules is discussed.     

Dynamics of detachment of Escherichia coli from plasma-mediated coatings under shear flow

A series of plasma-mediated coatings, containing silver nanoparticles embedded in an organosilicon or silica-like matrix, were deposited onto stainless steel and chemically characterized. Their anti-adhesive properties were evaluated in vitro towards Escherichia coli by performing shear-flow induced detachment experiments. Increasing the wall shear stress facilitated E. coli cell detachment, irrespective of the coating characteristics. When nanosilver was incorporated, cell detachment was lower, probably due to the affinity of the embedded silver for biological components of the cell wall. The presence of methyl groups in the matrix network could also promote enhanced hydrophobic interactions. Within the population fraction remaining attached to the coating under increasing shear flow, different association phenotypes were observed, viz. progressively lying flat, moving laterally, remaining tethered, or rotating by a single anchoring point, until alignment with the flow direction. This re-orientation phenotype and its relation with detachment were dependent of the coating. The effects of such heterogeneities should be more deeply explored.     

Critical stresses for cancer cell detachment in microchannels

We present experiments involving cancer cells adhering to microchannels, subjected to increasing shear stresses (0.1–30 Pa). Morphological studies were carried out at different shear stresses. Cells exhibit spreading patterns similar to those observed under static conditions, as long as the shear stress is not too high. At critical wall shear stresses (around 2−5 Pa), cell-substrate contact area decreases until detachment at the larger stresses. Critical shear stresses are found to be lower for higher confinements (i.e. smaller cell height to channel height ratio). Fluorescent techniques were used to locate focal adhesions (typically 1 μm² in size) under various shearing conditions, showing that cells increase the number of focal contacts in the region facing the flow. To analyze such data, we propose a model to determine the critical stress, resulting from the competition between hydrodynamic forces and the adhesive cell resistance. With this model, typical adhesive stresses exerted at each focal contact can be determined and are in agreement with previous works.     

Perfused transcapillary smooth muscle and endothelial cell co-culture—a novelin vitro model

Summary As mostin vitro endothelial cell (EC)-vascular smooth muscle cell (SMC) co-culture studies have been performed utilizing static culture conditions, none have successfully mimicked the physical environment of these cellsin vivo. EC covering the inner surface of blood vessels are continuously exposed to a hemodynamically imposed mechanical stress resulting from the flow of blood, while SMC are affected by pressure, a flow-related force acting perpendicular to the surface. We have developed a perfused transcapillary co-culture system that permits the chronic exposure of EC and SMC to physiological shear stresses and pressures. SMC and EC co-cultures were successfully established and maintained in long-term culture (7 wk) on an enclosed perfused bundle of semipermeable polypropylene capillaries. By altering flow rate and/or viscosity, shear stresses of 0.07–20 dyn/cm² can be readily achieved in this system. Electron microscopic analysis revealed that SMC formed multilayers around the outside of the capillaries, whereas EC, subjected to 3 dyn/cm² shear stress, formed an intact closely adherent monolayer lining the capillary lumen. EC and SMC exhibited characteristic ultrastructural and gross morphology. EC were separated from SMC by the capillary wall (pore size 0.5 μm, width 150 μM) and while no direct cell-cell contact was evident some cells were seen to migrate into the capillary wall. Both EC and SMC are exposed to the same culture medium, allowing the interaction of substances released in both directions. Yet separate populations of cells are maintained and can be individually harvested for further analysis. This co-culture system that mimics the architecture and physical environment of the vessel wall should have many potential applications in vascular biology.     

Immobilization of fungal spores by adhesion

Immobilization of conidiospores of Phanerochaete chrysosporium by adhesion was investigated in static and flow conditions on flat and on porous supports. Reducing the electrostatic repulsion between the spores and the support by adsorption of polycations on the support allows a better adhesion efficiency and a higher density of adhering spores and does not affect germination and growth. Formation of spore aggregates either in the suspension (high ionic strength) or on the support tends to decrease the surface coverage and to give an inhomogeneous distribution of adhering spores due to detachment of aggregates. The density of spores adhering from a flowing suspension is lower as compared with static conditions and does not exceed about 2% of surface coverage; this is due to the influence of tangential forces, to the short contact time with the surface, and to perturbation of the hydrodynamics along the surface by the previously immobilized spores. Obtaining a high coverage of the support by immobilized spores requires the absence of a tangential motion. (c) 1995 John Wiley & Sons, Inc.     

Shear‐induced detachment of biofilms from hollow fiber silicone membranes

A suite of techniques was utilized to evaluate the correlation between biofilm physiology, fluid‐induced shear stress, and detachment in hollow fiber membrane aerated bioreactors. Two monoculture species biofilms were grown on silicone fibers in a hollow fiber membrane aerated bioreactors (HfMBR) to assess detachment under laminar fluid flow conditions. Both physiology (biofilm thickness and roughness) and nutrient mass transport data indicated the presence of a steady state mature biofilm after 3 weeks of development. Surface shear stress proved to be an important parameter for predicting passive detachment for the two biofilms. The average shear stress at the surface of Nitrosomonas europaea biofilms (54.5 ± 3.2 mPa) was approximately 20% higher than for Pseudomonas aeruginosa biofilms (45.8 ± 7.7 mPa), resulting in higher biomass detachment. No significant difference in shear stress was measured between immature and mature biofilms of the same species. There was a significant difference in detached biomass for immature vs. mature biofilms in both species. However, there was no difference in detachment rate between the two species. Biotechnol. Bioeng. 2013; 110: 525–534. © 2012 Wiley Periodicals, Inc.