Physicochemical Characteristics and In Vitro Toxicity/Anti-SARS-CoV-2 Activity of Favipiravir Solid Lipid Nanoparticles (SLNs)

The rise of coronavirus (COVID-19) cases worldwide has driven the need to discover and develop novel therapeutics with superior efficacy to treat this disease. This study aims to develop an innovative aerosolized nano-formulation of favipiravir (FPV) as an anti-viral agent against coronavirus infection. The local delivery of FPV nanoparticles (NPs) via nebulization ensures that the drug can reach the site of infection, the lungs. Solid lipid NPs of favipiravir (FPV-SLNs) were formulated utilizing the hot-evaporation method. The physicochemical formulation properties were evaluated using dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The aerosol formulation performance was evaluated using an Andersen Cascade Impactor (ACI) at a flow rate of 15 L/min. The FPV-SLN formulation’s in vitro anti-viral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was also evaluated using the SARS-CoV-2 pathogen (hCoV-19/Egypt/NRC-3/2020 isolate). The FPV-SLNs’ morphology was defined utilizing transmission electron microscopy, showing an irregular shape. By means of FPV-SLNs’ nebulization, a fine particle fraction of 60.2 ± 1.7% was produced with 60.2 ± 1.7%, and this finding suggests that FPV-SLNs were appropriate for inhalation drug delivery with a particle size of 537.6 ± 55.72 nm. Importantly, the FPV-SLNs showed anti-viral activity against SARS-CoV-2 with CC50 and IC50 values of 449.6 and 29.9 µg/mL, respectively. This study suggests that inhaled solid lipid NPs of favipiravir could potentially be used against coronavirus.

Small quantities of respiratory syncytial virus RNA only in large droplets around infants hospitalized with acute respiratory infections

Background Respiratory syncytial virus (RSV) is a major cause of respiratory tract infections in young children. The predominant transmission routes for RSV are still a matter of debate. Specifically, it remains unclear if RSV can be transmitted through the air and what the correlation is between the amount of RSV in nasopharynx samples and in the air. Methods The amount of RSV in the air around hospitalized RSV infected infants in single-patient rooms was quantified using a six-stage Andersen cascade impactor that collects and fractionates aerosols and droplets according to size. RSV shedding in the nasopharynx of patients was followed longitudinally by quantifying RSV RNA levels and infectious virus in nasopharyngeal aspirates. Nose and throat swabs of parents and swabs of the patient’s bedrail and a datalogger were also collected. Results Patients remained RSV positive during the air sampling period and infectious virus was isolated up to 9 days post onset of symptoms. In three out of six patients, low levels of RSV RNA, but no infectious virus, were recovered from impactor collection plates that capture large droplets > 7 μm. For four of these patients, one or both parents were also positive for RSV. All surface swabs were RSV-negative. Conclusions Despite the prolonged detection of infectious RSV in the nasopharynx of patients, only small amounts of RSV RNA were collected from the air around three out of six patients, which were primarily contained in large droplets which do not remain suspended in the air for long periods of time.

Staphylococci and fecal bacteria as bioaerosol components in animal housing facilities in the Zoological Garden in Chorzów

Zoos are places open for a large number of visitors, adults and children, who can admire exotic as well as indigenous animal species. The premises for animals may contain pathogenic microbes, including those exhibiting antibiotic resistance. It poses a threat to people remaining within the zoo premises, both for animal keepers who meet animals on a daily basis and visitors who infrequently have contact with animals. There are almost no studies concerning the presence on the concentration of airborne bacteria, especially staphylococci and fecal bacteria in animal shelters in the zoo. There is no data about antibiotic resistance of staphylococci in these places. The results will enable to determine the scale of the threat that indicator bacteria from the bioaerosol pose to human health within zoo premises. This study conducted in rooms for 5 animals group (giraffes, camels, elephants, kangaroos, and Colobinae (species of monkey)) in the Silesian Zoological Garden in Chorzów (Poland). The bioaerosol samples were collected using a six-stage Andersen cascade impactor to assess the concentrations and size distribution of airborne bacteria. Staphylococci were isolated from bioaerosol and tested for antibiotic resistance. In our study, the highest contamination of staphylococci and fecal bacteria was recorded in rooms for camels and elephants, and the lowest in rooms for Colobinae. At least 2/3 of bacteria in bioaerosol constituted respirable fraction that migrates into the lower respiratory tract of the people. In investigated animal rooms, the greatest bacteria contribution was recorded for bioaerosol fraction sized 1.1–3.3μm. Bacterial concentrations were particularly strong in spring and autumn, what is related to shedding fur by animals. Among the isolated staphylococci which most often occurred were Staphylococcus succinus, S. sciuri, and S. vitulinus. The highest antibiotic resistance was noted in the case of Staphylococcus epidermidis, while the lowest for S. xylosus. In addition to standard cleaning of animal rooms, periodic disinfection should be considered. Cleaning should be carried out wet, which should reduce dust, and thus the concentrations of bacteria in the air of animal enclosures.

Health Risks Associated with Occupational Exposure to Biological Air Pollutants Occurring during the Processing of Biomass for Energy Purposes: A Case Study

The main goal of this study was to assess the levels of concentration of biological air pollutants in biomass processing power plants, based on the measurements taken in a plant located in Southern Poland. This work shows problems related to the co-combustion of biomass and indicates the health risks to which employees of positions closely related to the processing and combustion of biomass are exposed. Bacterial and fungal aerosol samples were collected using a six-stage Andersen cascade impactor with aerodynamic cut-off diameters of 7.0, 4.7, 3.3, 2.1, 1.1 and 0.65 µm (Tisch Environmental, USA). The highest average concentration level of micro-organisms was recorded in the air in the biomass analysis laboratory, where there was no constant airflow. It was found that the concentration levels of biological agents were below the Polish proposals for threshold limit values in the work environment polluted with organic dust. However, it was observed that the share of the respirable fraction (RF) of bacterial aerosol (particles less than 3.3 µm) in workplaces was equal to or higher than 50% of the total concentration, which may cause increased potentially adverse health effects and problems with concentration among power plant employees. Therefore, to ensure adequate air quality in the tested object, it is necessary to properly ventilate. This preliminary research could be the important starting point for a complete and precise assessment of bioaerosol exposures in biomass processing power plants in order to protect workers’ health.

Airborne Cupressaceae Pollen and Its Major Allergen, Cup a 1, in Urban Green Areas of Southern Iberian Peninsula

Cupressaceae pollen is responsible for winter respiratory allergies in the Mediterranean area. Pollen grains of this diverse family share the same characteristics under light microscopy. Consequently, the partial contribution of each Cupressaceae species to the airborne pollen spectrum cannot be determined with conventional aerobiological methods. Studies on major aeroallergens offer better information on the allergic sensitization and appearance of symptoms in patients sensitized to airborne pollen and spores. Our aim was to determine if airborne Cupressaceae pollen concentrations correspond to aerodynamics of the major allergen of Cupressus (Cup a 1), as well as the aeroallergen correlation with different-sized particles. The air was sampled in two cities of Southern Spain (Granada and Córdoba) using the Hirst-type volumetric sampler for airborne pollen detection during 2006 to 2008 and the Andersen Cascade Impactor for aeroallergen detection during the main pollination period of Cupressaceae in the same period. The samples were analyzed by the methodology proposed by the Spanish Aerobiology Network (REA), the minimum requirements of the European Aeroallergen Society (EAS) for pollen, and by ELISA immunoassay to detect airborne Cup a 1. The distribution patterns of airborne Cupressaceae pollen and Cup a 1 were observed throughout the sampling period, although with some irregular oscillations. Cupressaceae pollen and Cup a 1 showed positive and significant correlation during the period of maximum concentrations (January to March). However, the results of this study showed that the period of exposure of aeroallergens depends on the Cupressus species and their abundance in cities. According to the allergy potential obtained, species like C. arizonica could release more allergens than other species like C. sempervirens.

Small Quantities of Respiratory Syncytial Virus RNA Only in Large Droplets Around Infants Hospitalized With Acute Respiratory Infections

Background: Respiratory syncytial virus (RSV) is a major cause of respiratory tract infections in young children. The predominant transmission routes for RSV are still a matter of debate. Specifically, it remains unclear if RSV can be transmitted through the air and what the correlation is between the amount of RSV in nasopharynx samples and in the air.Methods: The amount of RSV in air around hospitalized RSV infected infants in single patient rooms was quantified using a six-stage Andersen cascade impactor that collects and fractionates aerosols and droplets according to size. RSV shedding in the nasopharynx of patients was followed longitudinally by quantifying RSV RNA levels and infectious virus in nasopharyngeal aspirates. Nose and throat swabs of parents and swabs of the patient’s bedrail and a datalogger were also collected.Results: Patients remained RSV positive during the air sampling period and infectious virus was isolated up to 9 days post onset of symptoms. In three out of six patients, low levels of RSV RNA, but no infectious virus, were recovered from impactor collection plates that capture large droplets >7 µm. For four of these patients, one or both parents were also positive for RSV. All surface swabs were RSV-negative.Conclusions: Despite the prolonged detection of infectious RSV in the nasopharynx of patients, only small amounts of RSV RNA were collected from air around three out of six patients, which were primarily contained in large droplets which do not remain suspended in air for long periods of time.

Staphylococci and Fecal Bacteria as Bioaerosol Components in Animal Housing Facilities in the Zoological Garden in Chorzów

Zoos are places open for a large number of visitors, adults and children, who can admire exotic as well as indigenous animal species. The premises for animals may contains pathogenic microbes, including those exhibiting antibiotic resistance. It poses a serious threat to people remaining within the zoo premises, both for animal keepers who meet animals on a daily basis and visitors who infrequently have contact with animals. Unfortunately, there are almost no studies concerning on the presense, on the concentration of airborne bacteria, especially staphylococci and faecal bacteria in animal shelters in the zoo. No data about antibiotic resistance of staphylococci in these places. The results will enable to determine the scale of the threat that indicator bacteria from the bioaerosol pose to human health within zoo premises. This study conducted in rooms for 5 animals group (giraffes, camels, elephants, kangaroos and colobinae) in the Silesian Zoological Garden in Chorzów (Poland). The bioaerosol samples were collected using a six-stage Andersen cascade impactor to assess the concentrations and size distribution of airborne bacteria. Staphylococci were isolated from bioaerosol and tested for antibiotic resistance. In our study the highest concentration of staphylococci and fecal bacteria was recorded in rooms for camels and elephants, the lowest in rooms for colobinae. At least 2/3 of bacteria in bioaerosol constituted respirable fraction, that migrates into the lower respiratory tract of the animals and the people. In investigated animal rooms the greatest bacteria contribution was recorded for bioaerosol fraction sized 1.1–3.3µm. Bacterial intoxication was particularly strong in spring and autumn, what is related to shedding fur by animals. Among the isolated staphylococci most often occurred Staphylococcus succinus, S. sciuri and S. vitulinus. The highest antibiotic resistance was noted in the case of Staphylococcus epidermidis, while the lowest for S. xylosus. As the animals constitute a significant source of staphylococci and fecal bacteria, attention should be paid to thorough cleaning of their shelters.

Pulmonary Targeting of Inhalable Moxifloxacin Microspheres for Effective Management of Tuberculosis

In the present study, the objective was to attain a localized lung delivery of an anti-tubercular fluoroquinolone, moxifloxacin (MXF), targeting the alveolar macrophages through a non-invasive pulmonary route using inhalable microspheres as a dry powder inhaler approach. MXF-loaded poly (lactic-co-glycolic acid) (PLGA) microspheres (MXF-PLGA-MSs) were fabricated by solvent evaporation technique and optimized by using a central composite statistical design. The morphology and particle size, as well as the flowability of the optimized microspheres, were characterized. In addition, the aerosolization performance of the optimized formula was inspected using an Andersen cascade impactor. Furthermore, in vivo fate following intrapulmonary administration of the optimized formula was evaluated. The optimized MXF-PLGA-MSs were spherical in shape with a particle size of 3.16 µm, drug loading of 21.98% and entrapment efficiency of 78.0%. The optimized formula showed a mass median aerodynamic diameter (MMAD) of 2.85 ± 1.04 µm with a favorable fine particle fraction of 72.77 ± 1.73%, suggesting that the powders were suitable for inhalation. Most importantly, in vivo studies revealed that optimized MXF-PLGA-MSs preferentially accumulated in lung tissue as manifested by a two-fold increase in the area under the curve AUC0–24h, compared to plain drug. In addition, optimized MXF-PLGA-MS sustained drug residence in the lung for up to 24 h following inhalation, compared to plain drug. In conclusion, inhalable microspheres of MXF could be a promising therapeutic approach that might aid in the effective eradiation of tuberculosis along with improving patient adherence to the treatment.

Effects of Bioaerosol Aerodynamic Size on Biodiversity in Three Kindergartens in Taiwan

Background: Human-associated bacteria (HAB) in the microbiome can become airborne bacterial aerosols (bioaerosols). The aerodynamic sizes of bioaerosols demonstrated may significantly affect their behaviors, respiratory deposition and biodiversity.Methods: The bacterial size, biodiversity and the HAB were evaluated at three kindergartens in central Taiwan in winter and spring. Kindergartens A, B, and C were in urban, semi-urban, and rural areas, respectively. A six-stage viable Andersen cascade impactor was used to collect bioaerosols and to determine their size distributions. A BD Phoenix-100 automated interpretation system was used to identify the species in those bioaerosols. The uniformity transformation was applied to verify the distributions of bacterial concentrations in different sampling rounds.Results: The results revealed 1,425 colonies (97.6%) that corresponded to 63 species in 29 genera and 35 colonies (2.4%) that were unidentified. The most abundant phylum was Actinobacteria (56.6+22.2%), Staphylococcus spp. was found in all sampling rounds, with a range of abundance between 2.0 and 70.0%. In all rounds, the geometric mean diameter of the bioaerosol and the geometric standard deviation of the bioaerosol size ranged from 2.19 to 5.42 µm and from 1.66 to 2.70, respectively. The Shannon diversity (H) and inverse Simpson diversity index (D) of the bioaerosols at each kindergarten were positively correlated with bioaerosol size, as larger bioaerosols had higher values of the biodiversity metrics. The Pearson correlations of H with the urbanization of the area of the kindergarten, temperature, RH and CO2 concentration were statistically significant (P < 0.05), indicating that the environmental variables and biodiversity covaried. The biodiversity in the rural area exceeded that in the urban area. Multiple and stepwise regression revealed that bioaerosol size was statistically significant with H (P = 0.001) and D (P = 0.002). The study may improve our understanding of the mechanisms and epidemiological spread of airborne infections.

Impact of bacterial aerosol, particulate matter, and microclimatic parameters on animal welfare in Chorzów (Poland) zoological garden

Zoos are very popular facilities visited by entire families with children, who come there to watch live animals. Zoos also provide workplaces for a large number of people directly looking after the animals. For places designed to house animals, regardless of whether they are farm animals, pets, or zoo animals, a higher concentration of both dust and potentially harmful bioaerosols can be expected. Unfortunately, there are almost no studies concerning the concentration of bacterial bioaerosols and particulate matter in animal shelters that would answer the question whether the level of these pollutants is constant or variable and dependent on a particular zoo, group of animals, their number in enclosures, or season. This study aimed to assess the levels of bacterial aerosol in rooms intended for animals (giraffes, camels, elephants, kangaroos, and colobinae) in the Silesian Zoological Garden in Chorzów (Poland). The bioaerosol samples were collected using a six-stage Andersen cascade impactor to assess the concentrations and size distribution of airborne bacteria. Particulate matter (PM10) was assessed using an electronic dust meter. Measurements of microclimate parameters were carried out using the Airflow™ Instruments Velocity Meter TA440, while gas concentrations were determined applying GFG Microtector II G450. The results showed that the concentration of airborne bacteria varied significantly between facilities for the analyzed animal groups. The lowest concentration of the total bacterial aerosol was observed in enclosures for colobinae (approx. 850 CFU/m3), while the highest—in rooms for elephants (approx. 105,600 CFU/m3). The average share of respirable fraction of bacteria was quite high, with values ranging from 62.9 (colobinae) to 86.9% (elephants), indicating potential harmfulness to the health of exposed people. PM10 concentrations were relatively low (10–86 μg/m3) and did not exceed the limit values for occupational exposure. Moreover, the levels of bacterial bioaerosol in almost all cases did not exceed the limit values. As the animals constitute a significant source of bioaerosol, attention should be paid to thorough cleaning of animals and their shelters, as well as maintaining appropriate levels of microclimate parameters in the facilities.