Passage of exogeneous fine particles from the lung into the brain in humans and animals.

There are still significant knowledge gaps in understanding the intrusion and retention of exogeneous particles into the central nervous system (CNS). Here, we uncovered various exogeneous fine particles in human cerebrospinal fluids (CSFs) and identified the ambient environmental or occupational exposure sources of these particles, including commonly found particles (e.g., Fe- and Ca-containing ones) and other compositions that have not been reported previously (such as malayaite and anatase TiO2), by mapping their chemical and structural fingerprints. Furthermore, using mouse and in vitro models, we unveiled a possible translocation pathway of various inhaled fine particles from the lung to the brain through blood circulation (via dedicated biodistribution and mechanistic studies). Importantly, with the aid of isotope labeling, we obtained the retention kinetics of inhaled fine particles in mice, indicating a much slower clearance rate of localized exogenous particles from the brain than from other main metabolic organs. Collectively, our results provide a piece of evidence on the intrusion of exogeneous particles into the CNS and support the association between the inhalation of exogenous particles and their transport into the brain tissues. This work thus provides additional insights for the continued investigation of the adverse effects of air pollution on the brain.

The effect of size and density on the mean retention time of particles in reindeer (Rangifer tarandus).

Particle passage from the reticulorumen (RR) depends on particle density and size. A classic way of assessing these effects is the use of plastic markers of varying density and size that are recovered in the faeces. Here, we report results of an experiment where four fistulated reindeer (Rangifer tarandus, 96 ± 12 kg) were fed two different diets (browse, voluntary dry matter intake [DMI] 70 ± 10 g/kg0.75/d; or a pelleted diet, DMI 124 ± 52 g/kg0.75/d) and dosed via fistula with 8 different particle types combining densities of 1.03, 1.22 and 1.44 g/ml and sizes of 1, 10 and 20 mm. Generally, particles that passed the digestive tract intact (not ruminated) did so relatively early after marker dosing, and therefore had shorter mean retention times (MRT) than ruminated particles. On the higher intake, the overall mean retention time (MRT) of particles was shorter, but this was not an effect of shorter MRT for either intact or ruminated particles, but due to a higher proportion of intact particles at the higher intake. This supports the concept that ruminants do not adjust chewing behaviour depending on intake, but that a lower proportion of digesta is submitted to rumination due to pressure-driven escape from the forestomach at higher gut fills. Compared to cattle (Bos primigenius taurus), muskoxen (Ovibos moschatus) and moose (Alces alces) that had received the same markers, reindeer had a lower proportion of 1 mm particles that passed intact. Our results support the concept that the critical size threshold for particles leaving the ruminant forestomach is dependent on body size. While the results likely do not represent findings peculiar for reindeer, they indicate fundamental mechanisms operating in the forestomach of ruminants.

Evaluating the mechanistic evidence and key data gaps in assessing the potential carcinogenicity of carbon nanotubes and nanofibers in humans.

In an evaluation of carbon nanotubes (CNTs) for the IARC Monograph 111, the Mechanisms Subgroup was tasked with assessing the strength of evidence on the potential carcinogenicity of CNTs in humans. The mechanistic evidence was considered to be not strong enough to alter the evaluations based on the animal data. In this paper, we provide an extended, in-depth examination of the in vivo and in vitro experimental studies according to current hypotheses on the carcinogenicity of inhaled particles and fibers. We cite additional studies of CNTs that were not available at the time of the IARC meeting in October 2014, and extend our evaluation to include carbon nanofibers (CNFs). Finally, we identify key data gaps and suggest research needs to reduce uncertainty. The focus of this review is on the cancer risk to workers exposed to airborne CNT or CNF during the production and use of these materials. The findings of this review, in general, affirm those of the original evaluation on the inadequate or limited evidence of carcinogenicity for most types of CNTs and CNFs at this time, and possible carcinogenicity of one type of CNT (MWCNT-7). The key evidence gaps to be filled by research include: investigation of possible associations between in vitro and early-stage in vivo events that may be predictive of lung cancer or mesothelioma, and systematic analysis of dose-response relationships across materials, including evaluation of the influence of physico-chemical properties and experimental factors on the observation of nonmalignant and malignant endpoints.

Computational modeling of nanoscale and microscale particle deposition, retention and dosimetry in the mouse respiratory tract.

Comparing effects of inhaled particles across rodent test systems and between rodent test systems and humans is a key obstacle to the interpretation of common toxicological test systems for human risk assessment. These comparisons, correlation with effects and prediction of effects, are best conducted using measures of tissue dose in the respiratory tract. Differences in lung geometry, physiology and the characteristics of ventilation can give rise to differences in the regional deposition of particles in the lung in these species. Differences in regional lung tissue doses cannot currently be measured experimentally. Regional lung tissue dosimetry can however be predicted using models developed for rats, monkeys, and humans. A computational model of particle respiratory tract deposition and clearance was developed for BALB/c and B6C3F1 mice, creating a cross-species suite of available models for particle dosimetry in the lung. Airflow and particle transport equations were solved throughout the respiratory tract of these mice strains to obtain temporal and spatial concentration of inhaled particles from which deposition fractions were determined. Particle inhalability (Inhalable fraction, IF) and upper respiratory tract (URT) deposition were directly related to particle diffusive and inertial properties. Measurements of the retained mass at several post-exposure times following exposure to iron oxide nanoparticles, micro- and nanoscale C60 fullerene, and nanoscale silver particles were used to calibrate and verify model predictions of total lung dose. Interstrain (mice) and interspecies (mouse, rat and human) differences in particle inhalability, fractional deposition and tissue dosimetry are described for ultrafine, fine and coarse particles.

Microplastic retention in marine vegetation canopies under breaking irregular waves.

The present study provides indications and underlying drivers of wave-induced transport and retention potential of microplastic particles (MP) in marine vegetation canopies having different densities. The anthropogenic occurrence of MP in coastal waters is well documented in the recent literature. It is acknowledged that coastal vegetation can serve as a sink for MP due to its energy dissipating features, which can mimic a novel ecosystem service. While the transport behavior of MP in vegetation has previously been investigated to some extent for stationary flow conditions, fundamental investigations for unsteady surf zone flow conditions under irregular waves are still lacking. Herein, we demonstrate by means of hydraulic model tests that a vegetation's retention potential of MP in waves increases with the vegetation shoot density, the MP settling velocity and decreasing wave energy. It is found that particles migrating by traction (predominantly in contact with the bed) are trapped in the wake regions around a canopy, whereas suspended particles are able to pass vegetated areas more easily. Very dense canopies can also promote the passage of MP with diameters larger than the plant spacing, as the canopies then show characteristics of a solid sill and avoid particle penetration. The particle migration ability through a marine vegetation canopy is quantified, and the key drivers are described by an empirical expression based on the particle settling velocity, the canopy length and density. The findings of this study may contribute to improved prediction and assessment of MP accumulation hotspots in vegetated coastal areas and, thus, may help in tracing MP sinks. Such knowledge can be considered a prerequisite to develope methods or new technologies to recover plastic pollutants and rehabilitate valuable coastal environments.

Biodiesel versus diesel exposure: enhanced pulmonary inflammation, oxidative stress, and differential morphological changes in the mouse lung.

The use of biodiesel (BD) or its blends with petroleum diesel (D) is considered to be a viable approach to reduce occupational and environmental exposures to particulate matter (PM). Due to its lower particulate mass emissions compared to D, use of BD is thought to alleviate adverse health effects. Considering BD fuel is mainly composed of unsaturated fatty acids, we hypothesize that BD exhaust particles could induce pronounced adverse outcomes, due to their ability to readily oxidize. The main objective of this study was to compare the effects of particles generated by engine fueled with neat BD and neat petroleum-based D. Biomarkers of tissue damage and inflammation were significantly elevated in lungs of mice exposed to BD particulates. Additionally, BD particulates caused a significant accumulation of oxidatively modified proteins and an increase in 4-hydroxynonenal. The up-regulation of inflammatory cytokines/chemokines/growth factors was higher in lungs upon BD particulate exposure. Histological evaluation of lung sections indicated presence of lymphocytic infiltrate and impaired clearance with prolonged retention of BD particulate in pigment laden macrophages. Taken together, these results clearly indicate that BD exhaust particles could exert more toxic effects compared to D.

Influence of exposure history on the particle retention capacity and physiological responses of Euonymus japonicus Thunb. var. aurea-marginatus Hort.

Green plants in urban environments experience cyclical particulate matter stress. And this history of exhaust exposure may generate stress memory in plants, which may alter their subsequent response. Studies combining urban pollution characteristics and stress memory are limited. Therefore, we selected E. japonicus var. aurea-marginatus, a common urban greening tree species in the Yangtze River Delta, and conducted an experiment in three periods: the initial pollution period (S1: 28 days), the recovery period (R: 14 days) and the secondary pollution period (S2: 28 days). The experimental design consisted of an elevated pollution treatment (173 µg•cm-3) and an ambient control (34 µg•cm-3) with three replicates. In S2, the net total particle retention and saturated particle retention decreased by 11.5% and 19.3%, respectively, while PM10 and PM2.5 did not change significantly. E. japonicus var. aurea-marginatus exhibited recovery of chlorophyll levels, slower degradation of carotenoid, faster accumulation of ASA, lower accumulation of MDA, reduced activity of SOD under the second pollution period, and the period had a significant effect on the physiological indicators. Collectively, the effect of autoexhaust exposure history on the particle retention capacity of selected plant varied across particle sizes, and stress memory may confer plant resistance to recurrent exhaust pollution via combined regulations of physiological responses. Fine particles which pose a great risk to human health arise predominantly from vehicular traffic and energy production. So, E. japonicus tends to play a stabilising role in particle retention in industrial, traffic and residential areas.

Particle uptake by filter-feeding macrofoulers from the Mar Grande of Taranto (Mediterranean Sea, Italy): potential as microplastic pollution bioremediators.

Microplastics (MPs) are a serious threat to the marine environment affecting ecosystem functioning and biodiversity. There is a vast literature about the uptake of MPs at different trophic levels, mainly focused on ecotoxicological effects in commercially relevant species. Little is still known about possible strategies to face MP pollution. Bioremediation is recently gaining attention in this framework. The clearance rate and particle retention of Sabella spallanzanii, Mytilus galloprovincialis, Phallusia mammillata, Paraleucilla magna at three MP concentrations (C1: 1.4 · 101 p/L; C2: 1.4 · 102 p/L; C3: 1.4 · 103 p/L) were investigated to test their potential as MP remover. Digestion protocol removed 98 % of tissues simplifying the MP quantification. P. magna clearance rate decreased with increasing concentration while P. mammillata showed no significant variations. S. spallanzanii and M. galloprovincialis instead exhibited the highest values of clearance rate. Yet, unlike mussels, S. spallanzanii can inhibit particle return to the surrounding water storing them in the tube, resulting to be the best candidate for bioremediation purposes.

A pore-scale investigation of microplastics migration and deposition during unsaturated flow in porous media.

Microplastics are ubiquitous in the natural environment and have the potential to endanger the natural environment, ecology and even human health. A series of microfluidic experiments by using soft lithography technology were carried out to investigate the effect of flow rate, particle volume fraction, particle size and pore/throat ratio on microplastics migration and deposition at the pore scale. We discovered a range of deposition patterns of the spherical microplastics from no particle deposition, to discontinuous particle layer, and to continuous particle layers in the retained liquid in the pores, depending on the particle size and volume fraction. Several metrics, including air saturation, probability of particle detainment, expansion ratio and thickness of residual liquid, were quantified to examine the role of various parameters on particle migration and retention of microplastics. At low flow rate (Q = 0.05 µL/min), microplastics migration and deposition were sensitive to changes in particle volume fraction, particle size and pore/throat ratio. In contrast, at high flow rates (Q > 5 µL/min), the migration and retention of particles were mainly controlled by strongly channelized air invasion pattern, while the particle volume fraction, particle size and pore/throat size ratio have only secondary influence. At intermediate range of flow rates, microplastics migration and deposition were dramatically impacted by flow rate, particle volume fraction, particle size and pore/throat ratio. This work improves the understanding of the mechanisms of particle migration and retention in porous media and can provide a reference for more accurate assessment of the exposure levels and times of microplastics in soil and groundwater systems.

Hydrodynamics and effect of velocity on particle filtration due to bridging in water-saturated porous media using CFD-DEM simulation.

Particle bridging owing to the confinement of the pore structure affects the transport and retention of particles in porous media. Particle motion driven by gravities were well investigated, whose filtration is mainly affected by the ratio of the particle diameter to the pore throat size of the medium. However, particles whose motions are driven by the fluid is essential to be investigated for particle separation from the carrying fluid. In this study, the motion of particles was driven by the liquid when passing through a water-saturated porous medium. The fluid-particle flow in a porous medium was modeled using computational fluid dynamics-discrete element method. The motion of particles in the slurry was traced in the porous medium, which enabled particle clogging to be directly precited by the interaction between the particles and pore surfaces by assessing the exact location of each particle. The pressure and flow field of the liquid were investigated, and the variation in flow path owing to particle clogging was predicted. The hydrodynamic study also showed that the Stokes number and particle concentration determined the particle clogging at the pore throats of the porous medium. Increasing the fluid velocity of particles such that the Stokes number was almost equal to 1 increased the separation efficiency of particles. Further increasing the fluid velocity reduced the residence time, which reduced the separation efficiency of the particles.

Macroalgal morphology mediates microplastic accumulation on thallus and in sediments.

The accumulation process of microplastics (MPs) is a key to understanding their fate in the environment. However, there is limited information about the short-term accumulation of MPs on macrophytes. The ability of macrophyte to attenuate wave and reduce current velocity is potentially facilitating MPs deposition. We hypothesize that the macroalgae retain MPs with their morphologies (filamentous and non-filamentous) being one of the factors to govern retention. Our hypothesis was tested by field observation during the dry season in Hong Kong when the macroalgae communities were the most diverse. MPs per biomass, surface area, or interstitial volume were used to represent the abundances on macroalgae. We found that filamentous algae retained a 2.35 times higher number of MPs when compared with non-filamentous algae if unit per biomass was considered. Other units, however, showed insignificant differences in MPs abundances between algal morphologies. Fibre was the most dominant shape of MPs with no significant difference in their abundances between filamentous and non-filamentous algae, suggesting fibres were retained regardless of the algal morphologies. To further evaluate the potential accumulation in the environment, sediment samples were also collected under the algal mat and immediate vicinity (~50 cm) of the algal mat. We found that sediment collected under the vegetated area contained significantly higher MPs. This was 3.39 times higher than the unvegetated area. Sediment collected under/near filamentous algae retained much higher abundances of MPs than those of non-filamentous algae. Provided that the observed retention of MPs on macroalgae, we speculate macrophyte system is one of the short-term MPs accumulation hotspots where the temporal increase of MPs depends on the seasonality of macrophyte in a given region.

The legacy effects of PM2.5 depositon on Nerium Oleander L.

Green plants have the capability to retain atmospheric particulate matter (PM) on their leaves, which can effectively reduce PM pollution, especially in the urban settings. Some studies reported that the periodic PM pollution could change plant retaining PM capacity, which, indeed, was the reason of physiological responses. In advancing the previous studies, we selected Nerium oleander L. to measure PM retention on leaf surface in a controlled environment by the following periods: initial pollution period (S1), recovery period (R), and secondary pollution period (S2) for a total of 12 weeks. The experimental design was one elevated pollution treatment (166 µg m-3) and one ambient control (28 µg m-3) with three replications. Results showed that during S2, the total retention decreased by 8.87 µg cm-2, which was about 10.4% significant lower than in S1. During the third week, the ascorbic acid content (ASA) in S1 was 6.71 mg g-1 significantly lower than that in S2 in the treatment. The total chlorophyll (Chl T) of the treatment decreased continuously and significantly by 33.8% in S1, but showed no similar trend in S2. The net photosynthetic rate of the treatment was significantly lower than that of the control, and the plants in the treatment showed a consistently high dark respiration rate than that in the control. The correlations between PM retention and ASA, Chl T and RWC were weaker in S1 than that in S2. In addition, air pollution tolerance index (APTI) showed a significant decline in plant pollution tolerance in the treatment during the third week.

Automobile exhaust particles retention capacity assessment of two common garden plants in different seasons in the Yangtze River Delta using open-top chambers.

Particulate matter (PM) pollution is a serious environmental problem in most of the cities in the Yangtze River Delta region. Plants can effectively filter ambient air by adsorbing PM. However, only a few studies have paid attention to the dynamic changes and seasonal differences in particle retention capacities of plants under long-term pollution. In this study, we investigated the dynamic changes in particle retention capabilities of the evergreen, broad-leaved, greening plants-Euonymus japonicus var. aurea-marginatus and Pittosporum tobira-in spring and summer. We employed an open-top chamber to simulate the severity of the tail gas pollution. The results showed that, both the plants reached a saturated state in 18-21 days, under continuous exposure to pollution (daily concentration of PM2.5: 214.64 ± 321.33 µg·cm-3). This was 6-8 days longer than that in the field experiments. In spring, the maximum retention of total particulate matter per unit leaf area of E. japonicus var. aurea-marginatus and P. tobira was 188.47 ± 3.72 µg cm-2 (18 days) and 67.63 ± 2.86 µg cm-2 (21 days), respectively. In summer, E. japonicus var. aurea-marginatus and P. tobira reached the maximum retention of the particle on the 21st day, with a net increase of 94.10 ± 3.77 µg cm-2 and 27.81 ± 3.57 µg cm-2, respectively. Irrespective of season, the particle retention capacity of E. japonicus var. aurea-marginatus was higher than that of P. tobira, and it showed a better effect on reducing the concentration of fine particles in the atmosphere. The particle retention of the two plants was higher in spring than that in summer. E. japonicus var. aurea-marginatus displayed a significant difference in particle retention between the seasons, while P. tobira did not show much difference. These results will provide a foundation for future studies on the dynamic changes and mechanism of particle retention in plants and management practices by employing plants for particle retention in severely polluted areas.

The role of Fe oxyhydroxide coating, illite clay, and peat moss in nanoscale titanium dioxide (nTiO2) retention and transport in geochemically heterogeneous media.

Natural media such as soil and sediment contain mineralogical and organic components with distinct chemical, surface, and electrostatic properties. To better understand the role of various soil and sediment components on particle transport, columns were packed with quartz sand and natural sediment with added Fe oxyhydroxide coating, illite clay, and peat moss to investigate how these added components influence nTiO2 retention and transport in geochemically heterogeneous medium. Results showed that nTiO2 transport was low at pH 5, attributable to the electrostatic attraction between positively-charged nTiO2 and negatively-charged medium. While illite did not notably affect nTiO2 transport at pH 5, Fe oxyhydroxide coating increased nTiO2 transport due to electrostatic repulsion between Fe oxyhydroxide and nTiO2. Peat moss also increased nTiO2 transport at pH 5, attributable to the increased DOC concentration, which resulted in higher DOC adsorption to nTiO2 and intensified electrostatic repulsion between nTiO2 and the medium. At pH 9, nTiO2 transport was high due to the electrostatic repulsion between negatively-charged nTiO2 and medium surfaces. Fe oxyhydroxide coating at pH 9 slightly delayed nTiO2 transport due to electrostatic attraction, while illite clay and peat moss substantially inhibited nTiO2 transport via straining/entrapment or electrostatic attraction. Overall, this study demonstrated that pH has a considerable effect on how minerals and organic components of a medium influence nTiO2 transport. At low pH, electrostatic attraction was the dominant mechanism, therefore, nTiO2 mobility was low regardless of the differences in mineralogical and organic components. Conversely, nTiO2 mobility was high at high pH and nTiO2 retention was dominated by straining/entrapment and sensitive to the mineralogical and organic composition of the medium.

Marker-ion analysis for quantification of mucoadhesivity of microparticles in particle-retention assays.

The objective of the present work was to develop an improved method to quantify particle retention on mucosal tissue under dynamic flow conditions with simultaneous determination of drug dissolution. The principle was to dissolve the collected inert carrier material and quantify specific marker ions by reliable analytical methods. The mucoadhesive model particles consisted of drug-loaded porous calcium carbonate microcarriers coated with chitosan, and quantification of calcium ions by capillary electrophoresis enabled to determine particle-retention kinetics on colonic mucosal tissue. The method was validated by image analysis, and the particle-retention assay was successfully applied to granulate material (125-250 mm) and small particles (<90 µm) with mucoadhesive properties. Particle retention on colonic mucosa was improved by increasing the chitosan content, demonstrating the sensitivity and usefulness of marker-ion analysis for quantification of detached particles. Furthermore, we showed that drug dissolution from mucoadhesive microparticles followed comparable kinetics in the particle-retention assay and the standard USP IV method. Our findings are helpful for the development of micro-sized colonic drug delivery systems, in particular for optimization of mucoadhesive properties and sustained drug release kinetics of porous drug carriers.

Influence of food size and food quantity on the feeding of the mussel Dreissena polymorpha.

In common with many other suspension feeders, the freshwater mussel Dreissena polymorpha has a maximum filtration rate at low food concentrations and a maximum ingestion rate at high food concentrations. These high rates, which reflect the potential maximum food uptake of the animal, are called the filtration capacity and the ingestion capacity respectively. The ingestion capacity was attained without forming pseudofaeces with Chlamydomonas reinhardii as food. The incipient limiting level could be calculated as the quotient of these two values. A decrease of the filtration rate at high food concentrations was correlated with changes in pumping activity, which showed more frequent interruptions, or a lower level of water transport. Dreissena can filter out particles of diameter greater than 0.7 µm from the water. Retention reaches a plateau at about 5 µm particle diameter. Scanning electron micrographs of the arrangement of the cilia on the gill filaments are given.

Faecal particle size distribution in captive wild ruminants: an approach to the browser/grazer dichotomy from the other end.

We investigated the particle size distribution in 245 faecal samples of 81 species of captive ruminants by a wet-sieving procedure. As a comparative measure, the modulus of fineness (MOF; Poppi et al. 1980) was used. Species were classified as frugivores (n=5), browsers (BR, n=16), intermediate feeders (IM, n=35) and grazers (GR, n=25). BR generally had a higher proportion of large particles, i.e. higher MOF values, than IM or GR of comparable size. These findings are in accord with reported lower fibre digestibility and less selective particle retention in BR, and are indicative of a difference in reticulo-ruminal physiology between the main ruminant feeding types. Possible consequences of the escape of larger particles from a browser's reticulo-rumen for the feeding of captive BR are briefly discussed.

Retention and remobilization dynamics of fine particles and microorganisms in pastoral streams.

Both microbial metabolism and pathogen retention and remobilization are dependent on downstream transport of fine particles, which migrate in a series of deposition and resuspension events. All fine particles, including clay minerals, particulate organic carbon, nutrients and microbes, are often considered to be transported similarly in the environment because of a lack of specific observations comparing their relative transport. We conducted a tracer injection study to compare the transport and retention of the fecal indicator bacterium Escherichia coli, synthetic inert fluorescent fine particles, and a dissolved conservative tracer. We found that the fluorescent fine particles and bacteria were transported similarly, with both having greater retention than the solute tracer. We used a stochastic model to evaluate in-stream retention and migration of the solute, fluorescent particles, and E. coli. The best-fit model parameters indicate that different stream reaches had varied retention characteristics, but always showed greater retention of fluorescent particles and E. coli compared to the solute tracer. Direct measurements within known retention areas after the injection showed that the majority of the fluorescent particles and E. coli were retained near the sediment-water interface in macrophyte stands or filtered within the top 3 cm of the streambed sediment. Both the tracer particles and E. coli were retained within these regions for multiple months following the injection experiment. The stochastic model properly captured the wide range of storage timescales and processes we observed in the stream. Our results demonstrate the importance of the streambed sediment and in-stream macrophytes as short- and long-term reservoirs for fine organic particles and microbes in streams.

Forage fiber effects on particle size reduction, ruminal stratification, and selective retention in heifers fed highly digestible grass/clover silages.

The objective of this study was to investigate the effect of NDF content in highly digestible grass/clover silage on particle size reduction, ruminal stratification, and selective retention in dairy heifers. The reduction in particle size from feed to feces was evaluated and related to feed intake, chewing activity, and apparent digestibility. Four grass/clover harvests (Mixtures of Lolium perenne, Trifolium pratense, and Trifolium repens) were performed from early May to late August at different maturities, at different regrowth stages, and with different clover proportions, resulting in silages with NDF contents of 312, 360, 371, and 446 g/kg DM, respectively, and decreasing NDF digestibility with greater NDF content. Four rumen-fistulated dairy heifers were fed silage at 90% of ad libitum level as the only feed source in a 4 × 4 Latin square design. Silage, ingested feed boluses, medial and ventral ruminal digesta, and feces samples were washed with neutral detergent in nylon bags of 10-µm pore size, freeze dried, and divided into small (<0.212 mm), medium (0.212 to 1 mm), and large : LP; >1 mm) particles by dry-sieving. Chewing activity, rumen pool size, and apparent digestibility were measured. Intake of NDF increased linearly from 2.3 to 2.8 kg/d with greater NDF content of forages (P = 0.01), but silages were exposed to similar eating time (P = 0.55) and rumination time per kg NDF (P = 0.35). No linear effect of NDF content was found on proportion of LP in ingested feed boluses (P = 0.31), medial rumen digesta (P = 0.95), ventral rumen digesta (P = 0.84), and feces (P = 0.09). Greater proportions of DM (P < 0.001) and particulate DM (P = 0.008) were found in medial ruminal digesta compared with ventral rumen, and differences in DM proportion increased with greater NDF content (P = 0.02). Particle size distributions were similar for digesta from the medial and ventral rumen regardless of NDF content of the silages (P > 0.13). The LP proportion was >30% of particles in the ventral and medial rumen, whereas in the feces, the LP proportion was <2%. Particle size stratification of the rumen was undetectable regardless of NDF content of the silages, stressing that the retention mechanism of large undigested particles lies elsewhere than with particle entrapment in the rumen mat. In this study, forage particle breakdown, ruminal stratification, and retention of particles in the rumen were not affected by NDF content of highly digestible grass/clover silages.