Ciliated Surface Research Articles

On non-Newtonian fluid flow generated via complex metachronal waves of cilia with magnetic, hall, and porous effects

Cilia beating influences bio-fluid flow, and conduits with ciliated surfaces serve numerous purposes. Cilia are hair-like adjuncts that produce liquid drive and cell locomotion. This novel research integrates magnetohydrodynamics, Hall effects, and porous medium with the cilia-driven flow. The developed problem is further simplified by utilizing famous assumptions, i.e., low Reynolds number and large wavelength. The bvp5c function in MATLAB (2023a) is adopted to achieve a convergent solution. The graphical illustrations of streamlines, pressure drop, and axial velocity under the influence of pertinent parameters are thoroughly examined. The findings suggest that when a high magnetic field (Hall current) and porous media are combined with the Carreau–Yasuda fluid, the pumping equipment operates better than the viscous liquid. In addition, fluid movement in biological organs is improved by metachronal ciliary motion. Furthermore, it is predicted that this study will make a significant contribution to biomedical engineering in terms of the upgrading and advancement of various types of drug-delivery technologies.

Impact of bio-mimetic cilia and symmetry breaking wave on double-diffusive radiated magneto Cu-Al2O3/bloodstream in an electroosmotic pump under momentum-thermal slip constraint

Transport of drug/hybrid materials through microfluidic channels requires efficient flow capability, effective mixing, and manipulation. To address this subject, a thermal analysis of double-diffusive stream of hybrid-type radiated bloodstream is considered through an elastic electroosmotic micropump in the presence of a magnetic field. Bio-mimetic cilia are embedded inside the pump surface having momentum-thermal slip effects. The inserted cilia strike and a constant phase difference between the beats of adjacent cilia generate a set of symmetry-breaking metachronal waves along the pump surface. The deemed mathematical model is simplified under the Debye–Hückel linearization and lubrication approach and is solved numerically. The analysis shows that the wall asymmetry and the apt cilia length strengthen the fluid stream and ensure active mixing distributed across significant distances. The radiated blood, the thin EDL, and extended cilia may be regarded as key factors in the cooling process. The entropy formation inside the micropump can be curtailed with the provision of an adequate amount of activation energy and the consideration of velocity-thermal slip effects on the ciliated surface. The intended model may deliver a deep perception of the thermal design of a futuristic drug delivery pump for efficient flow, adequate mixing, and least entropy generation.

Ciliated epithelia are key elements in the recruitment of bacterial partners in the squid-vibrio symbiosis.

The Hawaiian bobtail squid, Euprymna scolopes, harvests its luminous symbiont, Vibrio fischeri, from the surrounding seawater within hours of hatching. During embryogenesis, the host animal develops a nascent light organ with ciliated fields on each lateral surface. We hypothesized that these fields function to increase the efficiency of symbiont colonization of host tissues. Within minutes of hatching from the egg, the host's ciliated fields shed copious amounts of mucus in a non-specific response to bacterial surface molecules, specifically peptidoglycan (PGN), from the bacterioplankton in the surrounding seawater. Experimental manipulation of the system provided evidence that nitric oxide in the mucus drives an increase in ciliary beat frequency (CBF), and exposure to even small numbers of V. fischeri cells for short periods resulted in an additional increase in CBF. These results indicate that the light-organ ciliated fields respond specifically, sensitively, and rapidly, to the presence of nonspecific PGN as well as symbiont cells in the ambient seawater. Notably, the study provides the first evidence that this induction of an increase in CBF occurs as part of a thus far undiscovered initial phase in colonization of the squid host by its symbiont, i.e., host recognition of V. fischeri cues in the environment within minutes. Using a biophysics-based mathematical analysis, we showed that this rapid induction of increased CBF, while accelerating bacterial advection, is unlikely to be signaled by V. fischeri cells interacting directly with the organ surface. These overall changes in CBF were shown to significantly impact the efficiency of V. fischeri colonization of the host organ. Further, once V. fischeri has fully colonized the host tissues, i.e., about 12-24h after initial host-symbiont interactions, the symbionts drove an attenuation of mucus shedding from the ciliated fields, concomitant with an attenuation of the CBF. Taken together, these findings offer a window into the very first interactions of ciliated surfaces with their coevolved microbial partners.

Schistosoma mansoni Fibroblast Growth Factor Receptor A Orchestrates Multiple Functions in Schistosome Biology and in the Host-Parasite Interplay.

Stem cells play significant roles in driving the complex life cycle of Schistosoma mansoni. Fibroblast growth factor (FGF) receptor A (SmFGFRA) is essential for maintaining the integrity of schistosome stem cells. Using immunolocalization, we demonstrated that SmFGFRA was distributed abundantly in germinal/stem cells of different S. mansoni life stages including eggs, miracidia, cercariae, schistosomula and adult worms. Indeed, SmFGFRA was also localized amply in embryonic cells and in the perinuclear region of immature eggs; von Lichtenberg’s layer and the neural mass of mature eggs; the ciliated surface and neural mass of miracidia; the tegument cytosol of cercariae, schistosomula and adult worms; and was present in abundance in the testis and vitellaria of adult worms of S. mansoni. The distribution pattern of SmFGFRA illustrates the importance of this molecule in maintaining stem cells, development of the nervous and reproductive system of schistosomes, and in the host-parasite interplay. We showed SmFGFRA can bind human FGFs, activating the mitogen activated protein kinase (MAPK) pathway of adult worms in vitro. Inhibition of FGF signaling by the specific tyrosine kinase inhibitor BIBF 1120 significantly reduced egg hatching ability and affected the behavior of miracidia hatched from the treated eggs, emphasizing the importance of FGF signaling in driving the life cycle of S. mansoni. Our findings provide increased understanding of the complex schistosome life cycle and host-parasite interactions, indicating components of the FGF signaling pathway may represent promising targets for developing new interventions against schistosomiasis.

Residual time of sinusoidal metachronal ciliary flow of non-Newtonian fluid through ciliated walls: fertilization and implantation.

The monitoring of the ciliated walls in the uterine tube has supreme importance in enhancing the sperm to reach the egg (capacitation processes), and at peristaltic ciliary flow has a more favorable residual time along the canal when compared to the peristaltic flow. Based on the importance of this study, a mathematical simulation of this process has been carried out by studying the behavior of a non-Newtonian magnetized fluid with a Darcy flow model with an oscillating wall having an internal ciliated surface. The governing equation is formed with Eyring-Powell fluid (tubal fallopian fluid) without using any approximations and solved using the Adomian analysis method. Using the vorticity formula, the components of the velocity function, pressure gradient, and stream function are obtained. The influence of relevant parameters is explained through diagramming and discussion. We also analyzed the residue time effects on the flow parameters. The results indicate that peristaltic ciliary flow has a more favorable residual time along the canal when compared to peristaltic flow.

Pendrin Mediates Bicarbonate Secretion and Enhances Cystic Fibrosis Transmembrane Conductance Regulator Function in Airway Surface Epithelia.

Bicarbonate facilitates mucin unpacking and bacterial killing; however, its transport mechanisms in the airways are not well understood. cAMP stimulates anion efflux through the cystic fibrosis (CF) transmembrane conductance regulator (CFTR; ABCC7) anion channel, and this is defective in CF. The anion exchanger pendrin (SLC26A4) also mediates HCO3- efflux and is upregulated by proinflammatory cytokines. Here, we examined pendrin and CFTR expression and their contributions to HCO3- secretion by human nasal and bronchial epithelia. In native tissue, both proteins were most abundant at the apical pole of ciliated surface cells with little expression in submucosal glands. In well-differentiated primary nasal and bronchial cell cultures, IL-4 dramatically increased pendrin mRNA levels and apical immunostaining. Exposure to low-Cl- apical solution caused intracellular alkalinization (ΔpHi) that was enhanced fourfold by IL-4 pretreatment. ΔpHi was unaffected by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS) or CFTR inhibitor CFTRinh-172, but was reduced by adenoviral shRNA targeting pendrin. Forskolin increased ΔpHi, and this stimulation was prevented by CFTRinh-172, implicating CFTR, yet forskolin only increased ΔpHi after pendrin expression had been induced by IL-4. The dependence of ΔpHi on pendrin suggests there is minimal electrical coupling between Cl- and HCO3- fluxes and that CFTR activation increases anion exchange-mediated HCO3- influx. Conversely, inducing pendrin expression increased forskolin-stimulated, CFTRinh-172-sensitive current by approximately twofold in epithelial and nonepithelial cells. We conclude that pendrin mediates most HCO3- secretion across airway surface epithelium during inflammation and enhances electrogenic Cl- secretion via CFTR, as described for other SLC26A transporters.

Pendrin Mediates Bicarbonate Secretion and Enhances CFTR Function in Airway Surface Epithelia

Bicarbonate facilitates mucin unpacking and bacterial killing however its transport mechanisms in the airways are not well understood. cAMP stimulates anion efflux through the CFTR (ABCC7) anion channel and this is defective in CF. The anion exchanger pendrin (SLC26A4) also mediates HCO3− efflux and is upregulated by proinflammatory cytokines. Here we examined pendrin and CFTR expression and their contributions to HCO3− secretion by human nasal and bronchial epithelia. In native tissue, both proteins were most abundant at the apical pole of ciliated surface cells with little expression in submucosal glands. In well‐differentiated primary nasal and bronchial cell cultures, IL‐4 dramatically increased pendrin mRNA levels and apical immunostaining. Exposure to low‐Cl− apical solution caused intracellular alkalinization (ΔpHi) that was enhanced 4‐fold by IL‐4 pretreatment. ΔpHi was unaffected by DIDS or CFTRinh‐172 but was reduced by adenoviral shRNA targeting pendrin. Forskolin increased ΔpHi and this stimulation was prevented by CFTRinh‐172 implicating CFTR, yet forskolin only increased ΔpHi after pendrin expression had been induced by IL‐4. The dependence of ΔpHi on pendrin suggests there is minimal electrical coupling between Cl− and HCO3− fluxes and that CFTR activation increases anion exchange‐mediated HCO3− influx. Conversely, inducing pendrin expression increased forskolin‐stimulated, CFTRinh‐172‐sensitive current by ~2‐fold in epithelial and non‐epithelial cells. We conclude that pendrin mediates most HCO3− secretion across airway surface epithelium during inflammation and enhances electrogenic Cl− secretion via CFTR as described for other SLC26A transporters.Support or Funding InformationSupport: Cystic Fibrosis Canada, Canadian Institutes of Health Research, Canada Foundation for InnovationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The model squid-vibrio symbiosis provides a window into the impact of strain- and species-level differences during the initial stages of symbiont engagement.

Among horizontally acquired symbioses, the mechanisms underlying microbial strain- and species-level specificity remain poorly understood. Here, confocal-microscopy analyses and genetic manipulation of the squid-vibrio association revealed quantitative differences in a symbiont's capacity to interact with the host during initial engagement. Specifically, dominant strains of Vibrio fischeri, 'D-type', previously named for their dominant, single-strain colonization of the squid's bioluminescent organ, were compared with 'S-type', or 'sharing', strains, which can co-colonize the organ. These D-type strains typically: (i) formed aggregations of 100s-1000s of cells on the light-organ surface, up to 3 orders of magnitude larger than those of S-type strains; (ii) showed dominance in co-aggregation experiments, independent of inoculum size or strain proportion; (iii) perturbed larger areas of the organ's ciliated surface; and, (iv) appeared at the pore of the organ approximately 4×s more quickly than S-type strains. At least in part, genes responsible for biofilm synthesis control the hyperaggregation phenotype of a D-type strain. Other marine vibrios produced relatively small aggregations, while an array of marine Gram-positive and -negative species outside of the Vibrionaceae did not attach to the organ's surface. These studies provide insight into the impact of strain variation on early events leading to establishment of an environmentally acquired symbiosis.

Large‐conductance Ca2+‐activated K+ channel activation by apical P2Y receptor agonists requires hydrocortisone in differentiated airway epithelium

Hydrocortisone (HC) is required for activation of large-conductance Ca2+ -activated K+ current (BK) by purinergic receptor agonists. HC reduces insertion of the stress-regulated exon (STREX) in the KCNMA1 gene, permitting protein kinase C (PKC)-dependent channel activation. Overlapping and unique purinergic signalling regions exist at the apical border of differentiated surface cells. BK channels localize in the cilia of surface cells. In the present study we investigated the role of hydrocortisone (HC) on uridine-5'-triphosphate (UTP)-stimulated ion transport in differentiated, pseudostratified epithelia derived from normal human bronchial basal cells. The presence of a UTP-stimulated, paxilline-sensitive large-conductance Ca2+ -activated K+ (BK) current was demonstrated in control epithelia but was not stimulated in epithelia differentiated in the absence of HC (HC0). Addition of the BK channel opener NS11021 directly activated channels in control epithelia; however, under HC0 conditions, activation only occurred when UTP was added after NS11021. The PKC inhibitors GF109203x and Gö6983 blocked BK activation by UTP in control epithelia, suggesting that PKC-mediated phosphorylation plays a permissive role in purinoceptor-stimulated BK activation. Moreover, HC0 epithelia expressed significantly more KCNMA1 containing the stress-regulated exon (STREX), a splice-variant of the α-subunit that displays altered channel regulation by phosphorylation, compared to control epithelia. Furthermore, BK channels as well as purinergic receptors were shown to localize in unique and overlapping domains at the apical membrane of ciliated surface cells. These results establish a previously unrecognized role for glucocorticoids in regulation of BK channels in airway epithelial cells.

Ciliomotor circuitry underlying whole-body coordination of ciliary activity in the Platynereis larva.

Ciliated surfaces harbouring synchronously beating cilia can generate fluid flow or drive locomotion. In ciliary swimmers, ciliary beating, arrests, and changes in beat frequency are often coordinated across extended or discontinuous surfaces. To understand how such coordination is achieved, we studied the ciliated larvae of Platynereis dumerilii, a marine annelid. Platynereis larvae have segmental multiciliated cells that regularly display spontaneous coordinated ciliary arrests. We used whole-body connectomics, activity imaging, transgenesis, and neuron ablation to characterize the ciliomotor circuitry. We identified cholinergic, serotonergic, and catecholaminergic ciliomotor neurons. The synchronous rhythmic activation of cholinergic cells drives the coordinated arrests of all cilia. The serotonergic cells are active when cilia are beating. Serotonin inhibits the cholinergic rhythm, and increases ciliary beat frequency. Based on their connectivity and alternating activity, the catecholaminergic cells may generate the rhythm. The ciliomotor circuitry thus constitutes a stop-and-go pacemaker system for the whole-body coordination of ciliary locomotion.

Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium: a simple methodological approach for ex vivo studies of primary ciliary dyskinesia

BackgroundThree-dimensional explant spheroid formation is an ex vivo technique previously used in studies of airway epithelial ion and water transport. Explanted cells and sheets of nasal epithelium form fully differentiated spheroids enclosing a partly fluid-filled lumen with the ciliated apical surface facing the outside and accessible for analysis of ciliary function.MethodsWe performed a two-group comparison study of ciliary beat pattern and ciliary beat frequency in spheroids derived from nasal airway epithelium in patients with primary ciliary dyskinesia (PCD) and in healthy controls. Nasal ciliary cells and sheets were removed on day 1 by nasal brush biopsy and analyzed with regard to ciliary beat pattern—and frequency using high-speed video imaging for standard reference values. Three-dimensional explant spheroid formation was initiated in the same individual on the same day by incubation of cells and sheets from a separate brush biopsy. Harvested spheroids were analyzed earliest possible and values of spheroid ciliary beat pattern and frequency were compared to the corresponding reference values from day 1.ResultsSpheroids formed fast in serum-free culture medium. Formation was successful in 15 out of 18 (82%) sampled individuals. Thus, formation was successful in seven healthy controls and eight PCD patients, while unsuccessful in 3 with PCD due to infection. Median (range) number of days in culture before harvesting of spheroids was 4 (1–5) in healthy versus 2 (1–5) in PCD. Spheroid ciliary beat pattern and frequency were unchanged compared to their corresponding day 1 standard reference values. Spheroid ciliary beat frequency discriminated highly significant between healthy controls (9.3 Hz) and PCD patients (2.4 Hz) (P < 0.0001). Survival of spheroids was 16 days in a single healthy person.ConclusionPatient-specific three-dimensional explant spheroid formation from a minimal invasive nasal brush biopsy is a feasible, fast and valid ex vivo method to assess ciliary function with potential of aiding the diagnosis of PCD. In addition, it may be a useful model in the investigation of pathophysiological aspects and drug effects in human nasal airway epithelium.

Sternaspidae (Annelida, Sedentaria) from Vietnam with description of three new species and clarification of some morphological features.

Five sternaspid species were found near Vietnam shores: Sternaspis britayevi sp. nov., S. costata von Marenzeller, 1879, S. nana sp. nov., S. papillosa sp. nov., and S. spinosa Sluiter 1882. Sternaspis britayevi is described from the shallow water in Vietnam inhabiting soft bottoms; it resembles S. spinosa described from Java and S. thorsoni Sendall & Salazar-Vallejo, 2013 described from the Persian Gulf, but differs in having a medially projected and markedly ribbed fan of the ventro-caudal shield and nearly parallel, distally widened and rounded branchial plates. Sternaspis nana sp. nov. is described from Nha Trang Bay; it differs from the other known species by the combination of the following characters: small size, evenly distributed micropapillae and regular rows of long cirriform papillae; posterior chaetal fascicles consisting of single thick chaeta; a ventral shield with smooth integument, without ribs and usually without concentric lines. Sternaspis papillosa sp. nov. is also described from Nha Trang Bay; it resembles S. africana Augener, 1918 and S. andamanensis Sendall & Salazar-Vallejo, 2013 by having similar ventro-caudal shields but differs by body papillation and details of the ventro-caudal shield. Based upon observations of different species some morphological features are clarified: 1) notochaetae are present in introvert chaetigers as delicate capillaries; 2) peg-chaetae are really a dense group of more than 100 thin individual chaetae, embedded in a fibrous matrix, and covered by a common sheath; 3) the pharynx is an eversible, lobed, axial non-muscular proboscis with a ciliated surface; 4) the body cavity is divided by three septa in the anterior body region, and there are no other septa; and 5) an eversible anal peduncle is confirmed, as has been shown by early taxonomists.

Long-range interactions, wobbles, and phase defects in chains of model cilia.

Eukaryotic cilia and flagella are chemo-mechanical oscillators capable of generating long-range coordinated motions known as metachronal waves. Pair synchronization is a fundamental requirement for these collective dynamics, but it is generally not sufficient for collective phase-locking, chiefly due to the effect of long-range interactions. Here we explore experimentally and numerically a minimal model for a ciliated surface: hydrodynamically coupled oscillators rotating above a no-slip plane. Increasing their distance from the wall profoundly affects the global dynamics, due to variations in hydrodynamic interaction range. The array undergoes a transition from a traveling wave to either a steady chevron pattern or one punctuated by periodic phase defects. Within the transition between these regimes the system displays behavior reminiscent of chimera states.

Hyaluronan Rafts on Airway Epithelial Cells

Many cells, including murine airway epithelial cells, respond to a variety of inflammatory stimuli by synthesizing leukocyte-adhesive hyaluronan (HA) cables that remain attached to their cell surfaces. This study shows that air-liquid interface cultures of murine airway epithelial cells (AECs) also actively synthesize and release a majority of their HA onto their ciliated apical surfaces to form a heavy chain hyaluronan (HC-HA) matrix in the absence of inflammatory stimuli. These matrices do not resemble the rope-like HA cables but occur in distinct sheets or rafts that can capture and embed leukocytes from cell suspensions. The HC-HA modification involves the transfer of heavy chains from the inter-α-inhibitor (IαI) proteoglycan, which has two heavy chains (HC1 and HC2) on its chondroitin sulfate chain. The transesterification transfer of HCs from chondroitin sulfate to HA is mediated by tumor necrosis factor-induced gene 6 (TSG-6), which is up-regulated in inflammatory reactions. Because the AEC cultures do not have TSG-6 nor serum, the source of IαI, assays for HCs and TSG-6 were done. The results show that AECs synthesize TSG-6 and their own heavy chain donor (pre-IαI) with a single heavy chain 3 (HC3), which are also constitutively expressed by human renal proximal tubular epithelial cells. These leukocyte adhesive HC3-HA structures were also found in the bronchoalveolar lavage of naïve mice and were observed on their apical ciliated surfaces. Thus, these leukocyte-adhesive HA rafts are now identified as HC3-HA complexes that could be part of a host defense mechanism filling some important gaps in our current understanding of murine airway epithelial biology and secretions.

Three-dimensional, three-vector-component velocimetry of cilia-driven fluid flow using correlation-based approaches in optical coherence tomography.

Microscale quantification of cilia-driven fluid flow is an emerging area in medical physiology, including pulmonary and central nervous system physiology. Cilia-driven fluid flow is most completely described by a three-dimensional, three-component (3D3C) vector field. Here, we generate 3D3C velocimetry measurements by synthesizing higher dimensional data from lower dimensional measurements obtained using two separate optical coherence tomography (OCT)-based approaches: digital particle image velocimetry (DPIV) and dynamic light scattering (DLS)-OCT. Building on previous work, we first demonstrate directional DLS-OCT for 1D2C velocimetry measurements in the sub-1 mm/s regime (sub-2.5 inch/minute regime) of cilia-driven fluid flow in Xenopus epithelium, an important animal model of the ciliated respiratory tract. We then extend our analysis toward 3D3C measurements in Xenopus using both DLS-OCT and DPIV. We demonstrate the use of DPIV-based approaches towards flow imaging of Xenopus cerebrospinal fluid and mouse trachea, two other important ciliary systems. Both of these flows typically fall in the sub-100 μm/s regime (sub-0.25 inch/minute regime). Lastly, we develop a framework for optimizing the signal-to-noise ratio of 3D3C flow velocity measurements synthesized from 2D2C measures in non-orthogonal planes. In all, 3D3C OCT-based velocimetry has the potential to comprehensively characterize the flow performance of biological ciliated surfaces.

Lessons learned in andrology: looking at the spermatozoa in context.

Lessons learned in andrology: looking at the spermatozoa in context.

The many-body reciprocal theorem and swimmer hydrodynamics

We present a reinterpretation and extension of the reciprocal theorem for swimmers, extending its application from the motion of a single swimmer in an unbounded domain to the general setting, giving results for both swimmer interactions and general hydrodynamics. We illustrate the method for a squirmer near a planar surface, recovering standard literature results and extending them to a general squirming set, to motion in the presence of a ciliated surface, and expressions for the flow field throughout the domain. Finally, we present exact results for the hydrodynamics in two dimensions which shed light on the near-field behaviour.

Approximate modelling of the leftward flow and morphogen transport in the embryonic node by specifying vorticity at the ciliated surface

AbstractIn this paper, we have developed an approximate method for modelling the flow of embryonic fluid in a ventral node. We simplified the problem as flow in a two-dimensional cavity; the effect of rotating cilia was modelled by specifying a constant vorticity at the edge of the ciliated layer. We also developed an approximate solution for morphogen transport in the nodal pit. The solutions were obtained utilizing the proper generalized decomposition (PGD) method. We compared our approximate solutions with the results of numerical simulation of flow caused by the rotation of 81 cilia, and obtained reasonable agreement in most of the flow domain. We discuss locations where agreement is less accurate. The obtained semi-analytical solutions simplify the analysis of flow and morphogen distribution in a nodal pit.

Active Ciliated Surfaces Expel Model Swimmers

Continually moving cilia on the surface of marine organisms provide a natural defense against biofouling. To probe the physical mechanisms underlying this antifouling behavior, we integrate the lattice Boltzmann and immersed boundary methods and undertake the first computational studies of the interactions between actuated, biomimetic cilia and a model swimmer. We find that swimmers are effectively "knocked away" from the ciliated surface through a combination of steric repulsion and locally fluctuating flows. In addition, the net flow generated by the collective motion of the entire ciliary array was important for significantly reducing the times spent by relatively slow swimmers near the surface. The results reveal that active ciliated layers can offer a means to resist a wide range of species with a single surface.

Ciliate–adenovirus interactions in experimental co-cultures of Euplotes octocarinatus and in wastewater environment

The aim of the present work was to study the dynamics of the interactions between human adenovirus and ciliates under both experimental and field conditions. Experimental co-cultures of the ciliated protozoan Euplotes octocarinatus and human adenovirus (HAdV) type 2 were established and virus internalization was investigated using nested PCR and direct immunofluorescence (IF). In addition, to study protozoa-virus interactions in the field, wild ciliates were isolated from active sludges of a wastewater treatment plant and analyzed for the presence of adenovirus using direct IF. In vitro experiments revealed HAdV type 2 inside Euplotes cells after 15min of contact and its persistence until at least 35 days post infection. In addition, our results showed the adsorption of adenovirus on the surface of wild ciliates. We conclude that HAdV is taken up by ciliates, however more studies are necessary in order to better investigate the mechanisms, the infectivity of internalized virus and the protective effects of internalization against disinfection.