Study of nebulization delivery of aerosolized fluorescent microspheres to the avian respiratory tract.

This study investigated the delivery of an aerosol of monodisperse microspheres to the respiratory tract of birds following aerosol exposure. Adult domestic pigeons (Columbia livia domestica, n = 5 birds per timed treatment) were exposed to an aerosol of fluorescent 1.0 microm diameter carboxylate microspheres for 0.5, 1, 2, or 4 hr. During the aerosolization period, the birds were free-standing in a plexiglass treatment chamber and the aerosol was delivered using a commercial nebulizer. Immediately following aerosol exposure, the birds were euthanatized and the carcasses were intravenously infused with a modified paraformaldehyde/gluteraldehyde fixative. Evaluation of microsphere distribution was performed using a stereoscopic microscope with an epifluorescent module. The results from this study revealed that the amount of aerosolized particles delivered using a commercial nebulizer was proportional to exposure periods. Aerosol exposure periods of 0.5 hr or 1 hr did not result in a readily observable distribution of 1.0 microm fluorescent microspheres to the cranial thoracic, caudal thoracic, or abdominal air sac membranes. This was partly attributed to the relatively low concentration of the individual monodisperse microspheres in the aerosolized suspension. The 2- and 4-hr exposure periods resulted in readily observable deposition of the 1.0 mirom fluorescent microspheres in the cranial thoracic, caudal thoracic, or abdominal air sac membranes, with the 4-hr exposure period resulting in the greatest number of particles on the membrane surfaces. For each of the exposure periods, there was individual animal variation regarding the distribution and relative number of spheres deposited. This study demonstrates the widespread deposition of particles that had an aerodynamic equivalent diameter of approximately 1 microm and provides a better understanding of particle deposition efficiency within the respiratory system following aerosol exposure in birds.

Mechanisms of particulate matter toxicity in neonatal and young adult rat lungs.

Particulate matter (PM*) has been associated with a variety of adverse health effects, primarily involving the cardiovascular and respiratory systems. Researchers continue to investigate biologic mechanisms that may explain how exposure to PM exacerbates or directly causes adverse effects. Particle composition may play a critical role in these effects. In this study we used a diffusion flame system to generate ultrafine iron, soot, and iron combined with soot particles and exposed young adult and neonatal rats to different compositions of these particles. Young adult rats inhaled all three PM compositions on three consecutive days for 6 hours per day. Exposure to soot PM at 250 microg/m3 or to iron PM at 57 microg/m3 demonstrated no adverse respiratory effects. However, we observed mild pulmonary stress when the iron concentration was increased to 90 microg/m3. The most striking effects resulted when the rats inhaled PM composed of iron (45 microg/m3) combined with soot particles (total mass 250 microg/m3). This type of exposure produced significant indicators of oxidative stress, signs of inflammation, and increases in the levels of cytochrome P450 isozymes in the lungs. Repeated three-day exposure of neonatal rats to soot and iron particles in the second and the fourth weeks of life produced significant oxidative stress (elevations in oxidized and reduced glutathione) and ferritin induction. Neonatal rats exposed to PM in the second week of life also had a subtle but significant cell proliferation reduction in the centriacinar regions of the lungs. These findings suggest that iron combined with soot PM can lead to changes in the respiratory tract not found with exposure to iron or soot PM alone at similar concentrations. Unique effects in the neonate suggest that age may play an important role in susceptibility to inhaled particles.

An aerosolized fluorescent microsphere technique for evaluating particle deposition in the avian respiratory tract.

The objective of this study was to examine the feasibility of using aerosolized fluorescent microspheres to examine particle distribution in the respiratory tract of birds following aerosol exposure. Adult domestic pigeons (Columbia livia domestica; n = 5 birds per microsphere size) were exposed to aerosolized monodispersed populations of various sized carboxylate microspheres (0.5, 1.0, 2.0, 3.0, 6.0, and 10.0 microm) for 30 min. For aerosol-exposure purposes, the birds were anesthetized with injectable anesthetics, intubated, and placed on positive-pressure ventilation using a mechanical ventilator. Immediately following aerosol exposure, the birds were euthanatized, and carcasses were preserved via intravenous infusion of modified paraformaldehyde/gluteraldehyde fixative (pH = 7.2 and 340 mOsm). Initial evaluation of microsphere distribution in air sacs (cranial and caudal thoracic and abdominal) and at the level of the ostia was performed using a stereoscopic microscope with an epifluorescent module. More detailed examination of the distribution of microspheres within the respiratory tract was achieved using a confocal scanning laser microscope with a krypton argon laser and a scanning electron microscope. The results from this study revealed that positive-pressure ventilation resulted in distribution of smaller sized fluorescent microspheres (sizes 1.0, 2.0, and 3.0 microm) throughout the pigeon's respiratory tracts, and these microspheres were in highest concentration in the secondary bronchi and ostia for all of the examined air sacs. The larger sized beads (6.0 and 10.0) were confined to the upper airway (trachea and primary bronchi). The results from this study allow for a better understanding of particle deposition following positive-pressure ventilation and aerosol exposure in birds.

Reduced lung cell proliferation following short-term exposure to ultrafine soot and iron particles in neonatal rats: key to impaired lung growth?

Particulate matter (PM) has been associated with a variety of negative health outcomes in children involving the respiratory system and early development. However, the precise mechanisms to explain how exposure to airborne particles may cause adverse effects in children are unknown. To study their influence on early postnatal development, a simple, laminar diffusion flame was used to generate an aerosol of soot and iron particles in the size range of 10 to 50 nm. Exposure of 10-day-old rat pups to soot and iron particles was for 6 h/day for 3 days. The lungs were examined following a single injection of bromodeoxyuridine (BrdU) 2 h prior to necropsy. Neonatal rats exposed to these particles demonstrated no effect on the rate of cell proliferation within terminal bronchioles or the general lung parenchyma. In contrast, within those regions arising immediately beyond the terminal bronchioles (defined as the proximal alveolar region), the rate of cell proliferation was significantly reduced compared with filtered air controls. These findings strongly suggest exposure to airborne particles during early neonatal life has significant direct effects on lung growth by altering cell division within critical sites of the respiratory tract during periods of rapid postnatal development. Such effects may result in altered growth in the respiratory system that may be associated with lifelong consequences.

Airborne particles of the california central valley alter the lungs of healthy adult rats.

Epidemiologic studies have shown that airborne particulate matter (PM) with a mass median aerodynamic diameter < 10 microm (PM10) is associated with an increase in respiratory-related disease. However, there is a growing consensus that particles < 2.5 microm (PM2.5), including many in the ultrafine (< 0.1 microm) size range, may elicit greater adverse effects. PM is a complex mixture of organic and inorganic compounds; however, those components or properties responsible for biologic effects on the respiratory system have yet to be determined. During the fall and winter of 2000-2001, healthy adult Sprague-Dawley rats were exposed in six separate experiments to filtered air or combined fine (PM2.5) and ultrafine portions of ambient PM in Fresno, California, enhanced approximately 20-fold above outdoor levels. The intent of these studies was to determine if concentrated fine/ultrafine fractions of PM are cytotoxic and/or proinflammatory in the lungs of healthy adult rats. Exposures were for 4 hr/day for 3 consecutive days. The mean mass concentration of particles ranged from 190 to 847 microg/m3. PM was enriched primarily with ammonium nitrate, organic and elemental carbon, and metals. Viability of cells recovered by bronchoalveolar lavage (BAL) from rats exposed to concentrated PM was significantly decreased during 4 of 6 weeks, compared with rats exposed to filtered air (p< 0.05). Total numbers of BAL cells were increased during 1 week, and neutrophil numbers were increased during 2 weeks. These observations strongly suggest exposure to enhanced concentrations of ambient fine/ultrafine particles in Fresno is associated with mild, but significant, cellular effects in the lungs of healthy adult rats.