Concentrations Of Culturable Bacteria Research Articles

Evaluation of pathogen spread risk from excavated landfill

Landfill is a huge pathogen reservoir and needs special attention. Herein, the distribution and spread risk of pathogen were assessed in excavated landfill scenario. The results show that landfill excavation will greatly increase the risk of environmental microbial contamination. The highest total concentration of culturable bacteria among landfill refuse, topsoil and plant leaves was found to be as high as 1010 CFU g−1. Total coliforms, Hemolytic bacteria, Staphylococcus aureus, Salmonella, Enterococci, and Fecal coliforms were detected in the landfill surrounding environment. Notably, pathogens were more likely to adhere to plant leaves, making it an important source of secondary pathogens. The culturable bacteria concentration in the air samples differed with the landfill zone with different operation status, and the highest culturable bacteria concentration was found in the excavated area of the landfill (3.3 × 104 CFU m−3), which was the main source of bioaerosol release. The distribution of bioaerosols in the downwind outside of the landfill showed a tendency of increasing and then decreasing, and the highest concentration of bioaerosols outside of the landfill (6.56 × 104 CFU m−3) was significantly higher than that in the excavated area of the landfill. The risk of respiratory inhalation was the main pathway leading to infection, whereas the HQin (population inhalation hazardous quotient) at 500 m downwind the excavation landfill was still higher than 1, indicating that the neighboring residents were exposed to airborne microbial pollutants. The results of the study provide evidence for bioaerosols control protective measures taken to reduce health risk from the excavated landfill.

Culturable and inhalable airborne bacteria in a semiunderground municipal wastewater treatment plant: Distribution, transmission, and health risk assessment

Airborne pathogens constitute a growing threat to global public health. Wastewater treatment plants (WWTPs) are important sources of airborne bacteria, which pose great health risks to the employee and nearby residents. In this study, the distribution, transmission and health risk of the airborne culturable and inhalable bacteria carried by PM2.5 in a semiunderground WWTP were evaluated. The concentrations of culturable bacteria in the air were 21.2–1431.1 CFU/m3, with the main contributions of primary and biological treatments. The relative abundances of culturable and total inhalable bacterial taxa were positively correlated (p < 0.05). However, certain bacteria, including Bacillus, Acinetobacter and Enterococcus, exhibited high reproductive capacity despite their low concentration in the air, suggesting that they can survive and regrow in suitable environments. Transmission modeling revealed that the concentrations of airborne bacteria exponentially decreased with distance from 18.67 to 24.12 copies /m3 at the source to 0.06–0.14 copies /m3 at 1000 m downwind. The risks of 8-h exposure in this WWTP except the outlet exceeded the reference value recommended by WHO, which were primarily dependent on P. aeruginosa, Salmonella, and E. coli. Management practices should consider improved controls for bioaerosols in order to reduce the risk of disease transmission.

Assessment of indoor bioaerosol exposure using direct-reading versus traditional methods—potential application to home health care

Home healthcare workers (HHCWs) can be occupationally exposed to bioaerosols in their clients’ homes. However, choosing the appropriate method to measure bioaerosol exposures remains a challenge. Therefore, a systematic comparison of existing measurement approaches is essential. Bioaerosol measurements with a real-time, fluorescence-based Wideband Integrated Bioaerosol Sensor (WIBS) were compared to measurements with four traditional off-line methods (TOLMs). The TOLMS included optical microscopic counting of spore trap samples, microbial cultivation of impactor samples, qPCR, and next-generation sequencing (NGS) of filter samples. Measurements were conducted in an occupied apartment simulating the environments that HHCWs could encounter in their patients’ homes. Descriptive statistics and Spearman’s correlation test were computed to compare the real-time measurement with those of each TOLM. The results showed that the geometric mean number concentrations of the total fluorescent aerosol particles (TFAPs) detected with the WIBS were several orders of magnitude higher than those of total fungi or bacteria measured with the TOLMs. Among the TOLMs, concentrations obtained with qPCR and NGS were the closest to the WIBS detections. Correlations between the results obtained with the WIBS and TOLMs were not consistent. No correlation was found between the concentrations of fungi detected using microscopic counting and any of the WIBS fluorescent aerosol particle (FAP) types, either indoors or outdoors. In contrast, the total concentrations detected with microbial cultivation correlated with the WIBS TFAP results, both indoors and outdoors. Outdoors, the total concentration of culturable bacteria correlated with FAP-type AC. In addition, fungal and bacterial concentrations obtained with qPCR correlated with FAP types AB and AC. For a continuous, high-time resolution but broad scope, the real-time WIBS could be considered, whereas a TOLM would be the best choice for specific and more accurate microbial characterization. HHCWs’ activities tend to re-aerosolize bioaerosols causing wide temporal variation in bioparticle concentrations. Thus, the advantage of using the real-time instrument is to capture those variations. This study lays a foundation for future exposure assessment studies targeting HHCWs.

Correction to: Seasonal and vegetational variations of culturable bacteria concentrations in air from urban forest parks: a case study in Hunan, China

Correction to: Seasonal and vegetational variations of culturable bacteria concentrations in air from urban forest parks: a case study in Hunan, China

Distribution Characteristics and Health Effects of Bioaerosols in Spring over Xi'an City

A large amount of fluffy caktins appears in spring in Xi'an that may cause air pollution and lead to health risks such as asthma. In this study, bioaerosol, PM2.5, and fluffy catkins were collected from different sample points (traffic site and campus site) in Xi'an in spring. The variations in bioaerosol, fluffy catkins, and the bacterial community structure were investigated using culture-dependent and high-throughput sequencing methods. The results showed that the concentration of culturable bacteria was significantly higher (P=0.027) at the traffic site. The concentration of culturable bacteria at the traffic site was 2.7 times that of fungi, whereas the concentration of culturable fungi at the campus site was 1.4 times higher than that of bacteria. The peak concentrations of culturable bacteria and fungi appeared at 08:00 a.m. The size distribution of culturable bacteria showed a bimodal pattern, whereas that of culturable fungi showed a unimodal distribution. Soil and vegetation were the main sources of atmospheric microorganisms (85.9%), and Proteobacteria was the most abundant phylum in both fluffy catkins and PM2.5, accounting for 91.3% (traffic site) and 99.1% (campus site) of the fluffy catkins. Actinobacteria, Firmicutes, Bacteroidetes, Cyanobacteria, and Deinococcus-Thermus were the dominant phyla in PM2.5. Some genera were opportunistic pathogen bacteria in the fluffy catkins, such as Enterobacter and Pseudomonas, which can lead to infection and diarrhea risks. These results could provide fundamental data on potential health risks of spring-borne bioaerosols.

The changing pattern of bioaerosol characteristics, source and risk under diversity brush aerator speed

Bioaerosols containing pathogens released from wastewater treatment plants (WWTP) may pose potential health risks to workers on-site and residents downwind. In this study, sampling points were set up in the wastewater treatment facility to investigate the generation pattern of bioaerosols in the aeration tank section. High-throughput sequencing was utilized to assay the intestinal bacteria population, while the health risks associated with airborne bacteria were estimated based on average daily dose rates. The contribution of wastewater to bioaerosols was evaluated using the traceability analysis. As the rotational speed increased from 200 rpm to 800 rpm, the concentration of culturable bacteria increased from 397 CFU/m3 to 1611 CFU/m3, the proportion of bacteria attached to particles with an aerodynamic diameter larger than 4.7 µm increased from 30.41% to 48.44%, and the Shannon index of air samples increased from 1.032485 to 1.282065. Microbial composition, sources, and health risks of bioaerosols also changed as the rotational speed increased. The results showed that the predominant bacteria in the air at 200 rpm were Bacillus (78.78%), Paenibacillus (11.77%) and Lysinibacillus (1.40%). When the rotating speed reached 800 rpm, the dominant bacteria became Bacillus (55.50%), Acinetobacter (31.01%), and Paenarthrobacter (13.17%). The contribution of the wastewater to bioaerosols increased from 46.49% to 65.10%, in which surface water was the main source of bioaerosols (34.64% on average). Although the contribution of bottom water was lower than that of surface water, its contribution increased more, from 15.36% to 29.31%. The health risk of bioaerosols was 1.28 × 10−2 on average, which increased with the increase of rotational speed. At the same exposure concentration, children (2.31 × 10−2) have a higher exposure risk than adults (7.67 × 10−3). This study is aimed at exploring the variation law of bioaerosols discharged from WWTP with oxidation ditch process and providing preliminary data for reducing its risk.

Seasonal and vegetational variations of culturable bacteria concentrations in air from urban forest parks: a case study in Hunan, China.

It is important to investigate the airborne bacterial air quality in urban forest parks as tree bacteriostasis practices are being increasingly advocated as measures to improve the air quality and public health in urban green spaces around the world. The aim of the study was to quantitatively investigate airborne culturable bacteria (ACB) concentration levels based on field measurements in every season in five selected forest communities and the uncovered space in an urban forest park, as well as the effects of several factors on the culturability of airborne bacteria. Results suggested that the airborne bacterial levels of all the forest communities reached the clean air quality standard with regard to the airborne bacteria content, with the highest concentration of ACB showing in the uncovered space (1658 ± 1298 CFU/m3) and the lowest showing in the mixed community (907 ± 567 CFU/m3). The temporal distribution analysis showed that the airborne bacteria were mostly concentrated in summer, as well as in the morning and afternoon. The bacteriostatic rates of the mixed community were significantly different with seasonal variation (p < 0.05). Spearman's correlations revealed that the concentration of ACB was significantly positively correlated with the season, wind speed (WS), temperature (T), ultraviolet light (UV), negative air ion (NAI), and total suspended particles (TSP) (p<0.05) but significantly negatively correlated with the forest community type (p < 0.05). Overall, the selection of tree species plays a key role in shaping the forest structure and improving air quality, and the urban forest highlights key priorities for future efforts toward a cleaner, healthier, and more diverse regional forest environment.

Indoor and outdoor airborne bacterial and fungal air quality in kindergartens: Seasonal distribution, genera, levels, and factors influencing their concentration

Kindergartens in developing countries are sensitive places for children owing to exposure to bioaerosols that promote a range of infections. This work aimed to determine the concentration of culturable bacteria and fungi aerosols in indoor and outdoor air of twelve kindergartens in Rasht, Iran. The mean concentrations of fungi versus bacteria in indoor air of kindergartens were 7 ± 6 CFU/plate/hr versus 42 ± 29 CFU/plate/hr, respectively, while the mean concentrations of fungi versus bacteria in outdoor air of the same kindergartens were 12 ± 8 CFU/plate/hr versus 24 ± 18 CFU/plate/hr, respectively. The findings revealed that 33% of the concentration of indoor bioaerosols (bacteria and fungi) and 8% of the concentration of outdoor bioaerosols in kindergartens were higher than the recommended value (40–84 CFU/plate/hr), indicating medium risk. The main bacteria species detected in indoor and outdoor air in kindergartens were Bacillus spp., Staphylococcus aureus, Micrococcus spp., Staphylococcus epidermidis, Staphylococcus saprophyticus, Enterococcus spp., and Streptococcus spp. The predominant genera of the airborne fungi isolated from indoor and outdoor air in kindergartens were Aspergillus terreus, Aspergillus flavus, Cladosporium spp., Penicillium spp., Rhodotorula spp., Ulocladium spp., and Alternaria spp. Indoor air quality of kindergartens qualified as posing a medium risk level, and strategies should be considered to remove bioaerosol emissions in these susceptible places. Hence, to reduce negative health effects of bioaerosols on children, it is important to have proper ventilation, air conditioning systems, minimal furniture and textile materials, and application of disinfectants.

Prevention of particulate matter and airborne culturable bacteria transmission between double-tunnel ventilation layer hen houses

Airborne bacteria and particulate matter (PM) emitted from animal houses may harm the environment and threaten on-farm and off-farm bio-security. Airborne transmission of PM and bacteria occurs more easily between double-tunnel ventilation layer hen houses with side-to-side arrangement due to the layout of inlet and outlet, and the accumulation of airborne contaminants in the intermediate region. This study practically applied shading nets for covering exhaust area and water disinfection for evaporative cooling system to prevent airborne transmission of PM and microbial aerosols between layer hen houses. Differences of PM and airborne culturable bacteria (CB) concentrations in control group and experimental group were used to evaluate the contaminants blocking effect of the treatment. PM and airborne microbial concentrations between houses were significantly higher than the upwind. Shading nets used as a barrier of PM and airborne bacteria had no significant effect on the reduction of inlet PM and airborne CB concentration of layer hen houses. The airborne CB concentration after evaporative cooling pads with water could be several times higher than that before. However, airborne CB concentration after the evaporative cooling pads with water disinfection was much lower than that before. Results indicated that shading net as a barrier of PM and airborne bacteria could hardly bring down contaminants concentrations near the inlet. Water disinfection for evaporative cooling system could largely reduce the inlet airborne CB concentration. Inlet air purification of animal houses is an effective way to prevent airborne microorganisms transmission.

Microorganisms Collected from the Surface of Freshwater Lakes Using a Drone Water Sampling System (DOWSE)

New tools and technology are needed to study microorganisms in freshwater environments. Little is known about spatial distribution and ice nucleation activity (INA) of microorganisms in freshwater lakes. We developed a system to collect water samples from the surface of lakes using a 3D-printed sampling device tethered to a drone (DOWSE, DrOne Water Sampling SystEm). The DOWSE was used to collect surface water samples at different distances from the shore (1, 25, and 50 m) at eight different freshwater lakes in Austria in June 2018. Water samples were filtered, and microorganisms were cultured on two different media types, TSA (a general growth medium) and KBC (a medium semi-selective for bacteria in the genus Pseudomonas). Mean concentrations (colony forming units per mL, or CFU/mL) of bacteria cultured on TSA ranged from 19,800 (Wörthersee) to 210,500 (Gosaulacke) CFU/mL, and mean concentrations of bacteria cultured on KBC ranged from 2590 (Ossiachersee) to 11,000 (Vorderer Gosausee) CFU/mL. There was no significant difference in sampling distance from the shore for concentrations of microbes cultured on TSA (p = 0.28). A wireless bathymetry sensor was tethered to the drone to map temperature and depth across the sampling domain of each of the lakes. At the 50 m distance from the shore, temperature ranged from 17 (Hinterer Gosausee, and Gosaulacke) to 26 °C (Wörthersee), and depth ranged from 2.8 (Gosaulacke) to 11.1 m (Grundlsee). Contour maps of concentrations of culturable bacteria across the drone sampling domain revealed areas of high concentrations (hot spots) in some of the lakes. The percentage of ice-nucleation active (ice+) bacteria cultured on KBC ranged from 0% (0/64) (Wörthersee) to 58% (42/72) (Vorderer Gosausee), with a mean of 28% (153/544) for the entire sample set. Future work aims to elucidate the structure and function of entire microbial assemblages within and among the Austrian lakes.

Inhalable dust as a marker of exposure to airborne biological agents in composting facilities

ObjectivesIndustrial composting is associated with high levels of worker exposure to bioaerosols. Measurement of airborne microorganisms and endotoxin is complex and the related cost is high. The objective was therefore to examine whether dust measurement could be used as a marker of exposure to bioaerosols in composting facilities. MethodsA dataset of 110 measurements carried out in eight sludge composting plants was explored. Mixed-effects models were constructed to explain between-site and within-site variability in concentration of endotoxin and culturable mesophilic bacteria, mesophilic moulds and thermophilic actinomycetes in air. Fixed-effects variables were inhalable dust concentration, the season, the outdoor/indoor location of sampling and the process area. ResultsThe level of dust was a highly significant determinant of concentration for all biological agents. Within-site variability was always larger than between-site variability. The proportion of within-site variability explained by determinants was 68%, 65%, 56% and 60% for endotoxin, bacteria, moulds and actinomycetes, respectively. Inclusion of dust in the final model resulted in an increase of 24, 20, 12 and 17 points of percentage within-site variability, respectively. Inclusion of season resulted in an increase of 9, 12, 12 and 15 points, respectively. Within-site variability was less influenced by outdoor/indoor location and process area, except for moulds. ConclusionDust was the factor that most influenced within-site variability in endotoxin and culturable bacteria concentration. Measurement of dust can efficiently assist decision making for prevention measures against endotoxin and bacteria in sludge composting plants. Our results are not as conclusive for actinomycetes and especially for moulds.

Diversity and Ice Nucleation Activity of Microorganisms Collected With a Small Unmanned Aircraft System (sUAS) in France and the United States.

Many microbes relevant to crops, domestic animals, and humans are transported over long distances through the atmosphere. Some of these atmospheric microbes catalyze the freezing of water at higher temperatures and facilitate the onset of precipitation. We collected microbes from the lower atmosphere in France and the United States with a small unmanned aircraft system (sUAS). 55 sampling missions were conducted at two locations in France in 2014 (an airfield in Pujaut, and the top of Puy de Dôme), and three locations in the U.S. in 2015 (a farm in Blacksburg, Virginia, and a farm and a lake in Baton Rouge, Louisiana). The sUAS was a fixed-wing electric drone equipped with a remote-operated sampling device that was opened once the aircraft reached the desired sampling altitude (40–50 meters above ground level). Samples were collected on agar media (TSA, R4A, R2A, and CA) with and without the fungicide cycloheximide. Over 4,000 bacterial-like colonies were recovered across the 55 sUAS sampling missions. A positive relationship between sampling time and temperature and concentrations of culturable bacteria was observed for sUAS flights conducted in France, but not for sUAS flights conducted in Louisiana. A droplet freezing assay was used to screen nearly 2,000 colonies for ice nucleation activity, and 15 colonies were ice nucleation active at temperatures warmer than −8°C. Sequences from portions of 16S rDNA were used to identify 503 colonies from 54 flights to the level of genus. Assemblages of bacteria from sUAS flights in France (TSA) and sUAS flights in Louisiana (R4A) showed more similarity within locations than between locations. Bacteria collected with sUAS on TSA in France and Virginia were significantly different across all levels of classification tested (P < 0.001 for class, order, family, and genus). Principal Coordinates Analysis showed a strong association between the genera Curtobacterium, Pantoea, and Pseudomonas from sUAS flights in Virginia, and Agrococcus, Lysinibacillus, and Paenibacillus from sUAS flights in France. Future work aims to understand the potential origin of the atmospheric microbial assemblages collected with sUAS, and their association with mesoscale atmospheric processes.

Performance of personal electrostatic bioaerosol sampler (PEBS) when collecting airborne microorganisms

We recently developed a new personal electrostatic bioaerosol sampler (PEBS) for determining exposures to airborne microorganisms. PEBS was shown to collect airborne non-biological particles with efficiencies approaching 80% while producing very low ozone concentrations. In this work, we analyzed the performance of this sampler when collecting airborne Bacillus atrophaeus bacterial cells and Penicillium chrysogenum fungal spores as a function of sampling flow rates (e.g., 10 and 20 L/min) and sampling time (e.g., 10, 60, and 240 min). The collected samples were analyzed using microscopy, adenosine triphosphate (ATP)-based bioluminescence, flow cytometry (Live/Dead test), and culture techniques. PEBS's physical and biological performance was compared against that of BioSampler (SKC Inc., Eighty Four, PA) when the samplers were operated at 10 and 12.5 L/min, respectively. PEBS achieved physical collection efficiency as high as 83%, and its physical performance in terms of measured bioaerosol concentration was better than that of BioSampler. In addition, a fraction of live microorganisms recovered by PEBS was not different from that of BioSampler. Compared to BioSampler, PEBS measured similar or higher concentrations of culturable bacteria, but lower concentrations of culturable spores. The airborne ATP concentration measured by PEBS was significantly higher than that measured by BioSampler. Overall, we show that PEBS is a viable and efficient technology to determine personal exposures to airborne microorganisms using multiple sample analysis techniques.

Intestinal bacteria in bioaerosols and factors affecting their survival in two oxidation ditch process municipal wastewater treatment plants located in different regions

Samples from two oxidation ditch process municipal wastewater treatment plants (MWTPs) (HJK and GXQ) in two regions of China were analysed for bacteria, particles, total organic carbon, and water-soluble ions in bioaerosols. Diversity and potential pathogen populations were evaluated by high-throughput sequencing. Bioaerosol sources, factors affecting intestinal bacterial survival, and the relationship between bioaerosols and water were analysed by Source tracker and partial least squares-discriminant, principal component, and canonical correspondence analyses. Culturable bacteria concentrations were 110–846 and 27–579 CFU/m3 at HJK and GXQ, respectively. Intestinal bacteria constituted 6–33% of bacteria. Biochemical reaction tank, sludge dewatering house (SDH), and fine screen samples showed the greatest contribution to bioaerosol contamination. Enterobacter aerogenes was the main intestinal bacteria (> 99.5%) in HJK and detected at each sampling site. Enterobacter aerogenes (98.67% in SDH), Aeromonas sp. (76.3% in biochemical reaction tank), and Acinetobacter baumannii (99.89% in fine screens) were the main intestinal bacteria in GXQ. Total suspended particulate masses in SDH were 229.46 and 141.6 μg/m3 in HJK and GXQ, respectively. Percentages of insoluble compounds in total suspended particulates decreased as height increased. The main soluble ions in bioaerosols were Ca2+, Na+, Cl-, and SO42-, which ranged from 3.8 to 27.55 μg/m3 in the MWTPs. Water was a main source of intestinal bacteria in bioaerosols from the MWTPs. Bioaerosols in HJK but not in GXQ were closely related. Relative humidity and some ions positively influenced intestinal bacteria in bioaerosols, while wind speed and solar illumination had a negative influence.

Remote collection of microorganisms at two depths in a freshwater lake using an unmanned surface vehicle (USV).

Microorganisms are ubiquitous in freshwater aquatic environments, but little is known about their abundance, diversity, and transport. We designed and deployed a remote-operated water-sampling system onboard an unmanned surface vehicle (USV, a remote-controlled boat) to collect and characterize microbes in a freshwater lake in Virginia, USA. The USV collected water samples simultaneously at 5 and 50 cm below the surface of the water at three separate locations over three days in October, 2016. These samples were plated on a non-selective medium (TSA) and on a medium selective for the genus Pseudomonas (KBC) to estimate concentrations of culturable bacteria in the lake. Mean concentrations ranged from 134 to 407 CFU/mL for microbes cultured on TSA, and from 2 to 8 CFU/mL for microbes cultured on KBC. There was a significant difference in the concentration of microbes cultured on KBC across three sampling locations in the lake (P = 0.027), suggesting an uneven distribution of Pseudomonas across the locations sampled. There was also a significant difference in concentrations of microbes cultured on TSA across the three sampling days (P = 0.038), demonstrating daily fluctuations in concentrations of culturable bacteria. There was no significant difference in concentrations of microbes cultured on TSA (P = 0.707) and KBC (P = 0.641) across the two depths sampled, suggesting microorganisms were well-mixed between 5 and 50 cm below the surface of the water. About 1 percent (7/720) of the colonies recovered across all four sampling missions were ice nucleation active (ice+) at temperatures warmer than −10 °C. Our work extends traditional manned observations of aquatic environments to unmanned systems, and highlights the potential for USVs to understand the distribution and diversity of microbes within and above freshwater aquatic environments.

Concentration and Size Distribution Characteristics of Culturable Bioaerosols at Various Air Quality Levels During Fall and Winter in Xi'an, China

The concentration and size distribution of culturable bacteria and fungi were studied in Xi'an city at various air quality levels. The culturable bioaerosols were collected by an Andersen bioaerosol aerosol sampler between Sept. 2014 and Jan. 2015. Principal component analysis and multiple linear regressions were applied to link the concentrations with meteorological conditions including ambient temperature and relative humidity, as well as the levels of air pollutants such as PM2.5, PM10, NO2, SO2, and O3. These measured results showed that the concentration of culturable bacteria and fungi were in the ranges of 97-1909 CFU·m-3 and 92-1737 CFU·m-3, respectively. The concentrations of culturable bioaerosols increased along with a deterioration in air quality. The size distribution of the bacteria migrated to coarse particles. Fungal aerosols showed a normal distribution at low pollution levels, while for a high levels, they preferenced fine particles. Results from the principal component analysis (PCA) indicate that the concentration of culturable bioaerosols is mainly influenced by haze, solar radiation, and relative humidity. Multiple linear regression analysis showed that bacterial aerosol concentrations are positively correlated with haze (P<0.05) and relative humidity, while no significant negative correlations with solar radiation exists. Fungal aerosol concentrations did not have significant positive correlations with haze, solar radiation, or relative humidity. The results of this study will provide basic data for evaluating the effects of bioaerosols on human health and the environment.

Seasonality in airborne bacterial, fungal, and (1→3)-β-D-glucan concentrations in two indoor laboratory animal rooms

AimsThe purpose of this study was to assess the temporal changes in the concentrations of bioaerosols in a laboratory mouse room (LMR) and laboratory rabbit room (LRR), and to determine...

Monitoring of bacterial pathogens at workplaces in power plant using biochemical and molecular methods

PurposeThe aim of this study was to characterize the ways of spreading of the most common bacterial species isolated from workers as well as from the air and raw materials at the workplaces in power plant utilizing biomass sources. To monitor microbial transmission and identify the source of contamination in the working environment, a combination of molecular and biochemical methods was applied.MethodsThe study was carried out at workplaces in power plant utilizes biomass as a main fuel source. At each of the studied workplaces, bioaerosol particles were collected on sterile Teflon filters using personal conical inhalable samplers (CIS), and biomass samples (straw pellets and briquettes, corn briquettes, sunflower pellets and wood chips) were directly taken from their storage places. Simultaneously with that, the swab samples from the hands of ten workers and their used respiratory masks (of FFP2 class) were also collected after the work shift to evaluate individual workers’ microbial contamination. In all collected samples, total bacterial concentrations were assessed and the most common microbial isolates were identified to the species level using both biochemical (API tests) and molecular polymerase chain reaction (PCR), followed by random amplification of polymorphic DNA (RAPD) typing methods.ResultsThe mean concentrations of culturable bacteria in the air and in biomass samples at the studied workplaces were high, i.e. 1.2 × 106cfu/m3 and 3.8 × 104cfu/g, respectively. The number of bacteria in the swab and mask samples also reached a high level of 1.4 × 104 cfu/ml and 1.9 × 103 cfu/cm2, respectively. Among the most frequently isolated microorganisms from all types of samples were Gram-positive bacteria of the genus Bacillus and Staphylococcus xylosus. 37 bacterial strains belonging to the genus Bacillus (B. licheniformis 8, B. pumilus 15 and B. subtilis 4) and Staphylococcus (10) were genotyped by the RAPD-PCR method. Based on RAPD-PCR analyses, the genomic similarity among 19 Bacillus strains isolated from biomass, air, protective mask and hand samples as well as 6 S. xylosus strains isolated from air, mask and hand samples exceeded 80%.ConclusionThis study demonstrated that biomass is the primary source of bacteria at power plant workplaces. These results also revealed that biomass-associated bacteria can be easily transferred to workers’ hands and mask during their routine activities. To improve health protection at the workplaces, adequate training courses on hand hygiene and how to use and remove respiratory masks correctly for workers should be introduced as a key element of the prevention strategy. From the occupational point of view, the PCR-based methods seem to be an efficient tool for a fast and precise typing of bacterial strains isolated from different sources in the occupational environment. Such methods may help to implement appropriate prophylactic procedures and minimize transmission of infectious agents at workplaces.

주택내 주거공간에 따른 부유세균 농도 분포 및 종 동정 연구

Objectives: Exposure to airborne bacteria is associated with adverse health effects such as respiratory and infectious diseases. This study evaluated airborne bacterial concentrations in the living rooms, kitchens, and toilets of 30 homes. Methods: Bacteria were sampled with an MAS100 impactor in three spaces in the subject homes between April 2014 and February 2015. Bacteria were grown on TSA plates for 48 hours at <TEX>$35^{\circ}C$</TEX>. The bacterial strains were isolated and amplified by polymerase chain reaction. Results: The most culturable bacteria were found in toilets (<TEX>$624.0CFU/m^3$</TEX>, GM: <TEX>$417.3CFU/m^3$</TEX>), followed by in the kitchen (<TEX>$503.8CFU/m^3$</TEX>, GM: <TEX>$324.9CFU/m^3$</TEX>). The dominant genera identified were: Staphylococcus sp.(19%), Micrococcus sp.(16%), and Bacillus sp.(11%) in the indoor air and Bacillus sp. (30%) in the outdoor air. Gram-positive bacteria comprised more than half of all colonies. Conclusion: In this study, culturable bacteria concentrations were higher than those reported in other spaces. Therefore, it is important to control relative humidity and remove moisture to prevent bacteria from multiplying. Additionally, the dominant species in indoor air were Staphylococcus sp. and Micrococcus sp. These are found on the human skin, mucous membranes, and hair, so human activity can affect bacterial distribution. Therefore, cleaning and controlling moisture are important for reducing indoor bacterial concentrations.

Optimization of slightly acidic electrolyzed water spray for airborne culturable bacteria reduction in animal housing

Slightly acidic electrolyzed water (SAEW) spray has been considered as a novel approach for airborne bacteria reduction in animal housing. This study aimed to optimize the operating parameters of SAEW spray based on the size distribution of sprayed aerosols, the available chlorine travelling loss in sprayed aerosols, and the reduction efficiency of airborne culturable bacteria (CB). The optimized operating parameters were the nozzle orifice diameter and the spray pressure. The size distribution characteristics of sprayed SAEW aerosols under different operating parameters were quantified. The pH and available chlorine concentrations of the original SAEW solution and the SAEW aerosols collected at 0 m, 0.25 m, 0.50 m, 0.75 m, and 1.00 m from the spray nozzle head were analyzed. A bioaerosol nebulizing generator was used to simulate animal housing environment in an environmentally-controlled chamber. Six SAEW spray trials under different operating parameters were conducted at a spray dosage of 80 mL/m2 in the chamber. Airborne CB concentrations before and after spray were measured to investigate the reduction efficiency of airborne CB. Dv(50), the volume-based diameter below which 50% of the particle being present, increased with the nozzle orifice diameter and decreased with the spray pressure. The travelling loss of available chlorine in the sprayed SAEW aerosols was greatly dependent on Dv(50). SAEW spray with medium size sprayed aerosols (Dv(50) = 86.62 μm, 67.94 μm, and 54.53 μm) showed significantly higher airborne CB reduction efficiencies than large (Dv(50)=121.80 μm and 96.00 μm) or small size aerosols (Dv(50) = 42.57 μm). The spray operating parameters that provide medium size sprayed aerosols (Dv(50) ~ 60-90 μm) are recommended for SAEW spray in animal housing. Keywords: animal housing, bioaerosol, available chlorine, size distribution, air quality, poultry and livestock DOI: 10.3965/j.ijabe.20160904.2366 Citation: Zheng W C, Ni L, Hui X, Li B M, Zhang J F. Optimization of slightly acidic electrolyzed water spray for airborne culturable bacteria reduction in animal housing. Int J Agric & Biol Eng, 2016; 9(4): 185－191.