Outdoor airborne fungi captured by viable and non-viable methods

Volume assessments of the concentration of airborne fungi may provide different results depending on the methodology used. This work simultaneously analyses two methods for samples obtained outdoors and analysed in the context of meteorological conditions. The study was carried out in Badajoz (SW Spain) from Mar. 2009 to Jul. 2011. A Burkard fixed spore trap was used for the non-viable sampling, and three different methods were used for the viable sampling: a Burkard portable spore trap with two inlet port types and a Sampl'air AES trap. Daily average total concentrations of 285 CFU m⁻³ and 1 954 spores m⁻³ were recorded for the viable and non-viable methods, respectively. The spore/colony ratio showed important differences among the most relevant fungal types: Alternaria (2.6), Aspergillus–Penicillium (2.0) and Cladosporium (11.1). Although the two sampling types were essentially equivalent at showing temporal variations in outdoor airborne fungi, quantitative differences in the number of total colonies recorded depended on the culture media and conditions used.     

Lateral fungal spore movement inside a simulated wall

Moisture inside walls can facilitate mold growth if left untreated. Once spores become airborne they may interact with pressures inside walls. Two laboratory experiments were conducted to determine if airborne spores have the potential to migrate laterally inside walls with and without wiring installations. A simulated wall was fabricated, and Penicillium chrysogenum spores were aerosolized into a distant stud bay and an adjacent stud bay. The wall was subjected to a typical indoor pressure. Spore levels inside the bays were sampled, and a total of 36 trials (n = 36) were conducted. Results of Kruskal–Wallis tests revealed that spore levels inside the sampling bay and the distant bay with wiring installations were not significantly different. Spore levels inside the sampling bay were significantly lower than the adjacent bay without wiring installations (P < 0.05). The findings of the study suggest airborne fungal spores have the potential to move laterally inside walls.     

Adaptations in muscular activation of the knee extensor muscles with strength training in young and older adults

The purpose of this study was to compare the extent of muscular activation during maximal voluntary knee extension contractions in old and young individuals and to examine the effects of resistance training on muscular activation in each group. The interpolated twitch technique was used to estimate muscular activation during two pre-training baseline tests, and after two and six weeks of resistance training. Throughout the study, the older group was 30% less strong than the young group (p=0.02). The training protocol was effective in both groups with overall isometric strength gains of 30 and 36% in the older (p=0.01) and young (p<0.01) groups, respectively. 10-RM training loads increased by 66% in the old group (p<0.01) and by 77% in the young group (p<0.01) throughout training. At the first baseline test, a 2% difference in muscular activation between groups (p=0.3) did not explain the large disparity in strength. Muscular activation increased by 2% in both groups throughout training (p<0.01). Despite considerably less muscular strength in the older group, muscular activation was greater than 95% of maximum and appears to be equal in both young and older individuals. Both groups demonstrated similar but small increases in muscular activation throughout training.     

Relationship between indices of iron status and coronary risk factors including diabetes and the metabolic syndrome in Saudi subjects without overt coronary disease

There have been inconsistent reports on the relationship between iron status and coronary artery diseases (CAD), and little data on this relationship in non-Caucasian populations.

We assessed dietary iron by questionnaire and measured serum iron and ferritin levels in 270 Saudi male subjects without established CAD, 130 of whom were angiogram negative. Serum lipid profile, glucose, high sensitivity-C reactive protein (hs-CRP), serum soluble intercellular adhesion molecules-1 (sICAM-1), and caeruloplasmin were measured in all subjects.

The angiogram negative patients, had lower serum ferritin (p<0.05) and iron (p<0.0001) levels than the 140 subjects without reported cardiovascular diseases (CVD). Serum iron correlated with serum triglycerides (p<0.0001) and total cholesterol (p<0.05) levels for this latter group and the groups combined. Serum ferritin correlated with serum total cholesterol and low-density lipoprotein (LDL)-cholesterol in the combined group (p<0.05), and was correlated with blood glucose and serum LDL-cholesterol (p<0.05) in the subjects without reported CVD. After adjustment for confounding variables, serum iron levels remained a significant correlate with total calorie intake and serum triglycerides. Serum ferritin also correlated significantly with cholesterol intake and fasting serum total cholesterol. Dietary iron was significantly related to dietary cholesterol and fiber, age, smoking habits, and serum total cholesterol level.

Hence, indices of iron status were related to several coronary risk factors in the Saudi population.     

Suitability of poor medium in counting total viable airborne bacteria

The purpose of this study was to test the effect of incubation temperature and culture medium on viable counts of airborne bacteria. The incubation temperature had different effect on indoor and outdoor air bacteria. Indoor air bacteria grew as well at 20°C as 37°C, but less at 10°C. Outdoor air bacteria grew equally well at 10°C and 20°C, but less at 37°C. Both indoor and outdoor air bacteria grew differently on poor and rich media. The counts of both indoor and outdoor air bacteria were higher on poor R2A medium (low nutrient concentration) than on rich TYG and blood media (high nutrient concentration). The results indicate that a poor medium incubated at 20°C is adequate for counting viable airborne bacteria.     

Real-time monitoring of non-viable airborne particles correlates with airborne colonies and represents an acceptable surrogate for daily assessment of cell-processing cleanroom performance

Background aimsAirborne particulate monitoring is mandated as a component of good manufacturing practice. We present a procedure developed to monitor and interpret airborne particulates in an International Organization for Standardization (ISO) class 7 cleanroom used for the cell processing of Section 351 and Section 361 products.MethodsWe collected paired viable and non-viable airborne particle data over a period of 1 year in locations chosen to provide a range of air quality. We used receiver operator characteristic (ROC) analysis to determine empirically the relationship between non-viable and viable airborne particle counts.ResultsViable and non-viable particles were well-correlated (r² = 0.78), with outlier observations at the low end of the scale (non-viable particles without detectable airborne colonies). ROC analysis predicted viable counts ≥ 0.5/feet³ (a limit set by the United States Pharmacopeia) at an action limit of ≥ 32 000 particles (≥ 0.5 µ)/feet³, with 95.6% sensitivity and 50% specificity. This limit was exceeded 2.6 times during 18 months of retrospective daily cleanroom data (an expected false alarm rate of 1.3 times/year). After implementing this action limit, we were alerted in real time to an air-handling failure undetected by our hospital facilities management.ConclusionsA rational action limit for non-viable particles was determined based on the correlation with airborne colonies. Reaching or exceeding the action limit of 32 000 non-viable particles/feet³ triggers suspension of cleanroom cell-processing activities, deep cleaning, investigation of air handling, and a deviation management process. Our full procedure for particle monitoring is available as an online supplement.     

Quantitative sampling of indoor air biomass by signature lipid biomarker analysis

Exposure to airborne biocontaminants may result in a multitude of health effects and is related to a pronounced increase in adult-onset asthma. Established culture-based procedures for quantifying microbial biomass from airborne environments have severe limitations. Assay of the phospholipid fatty acid (PLFA) components of airborne microorganisms provides a quantitative method to define biomass, community composition and nutritional/physiological activity of the microbial community. By collecting airborne particulate matter from a high volume via filtration, we collected sufficient biomass for quantitative PLFA analysis. Comparing high (filtration) and low (impaction) volume air sampling techniques at 26 locations within the Eastern United States, we determined that PLFA analysis provided a viable alternative to the established but flawed culture-based techniques for measuring airborne microbial biomass and community composition. Compared to the PLFA analysis, the culture techniques underestimated the actual viable airborne biomass present by between one to three orders of magnitude. A case study of a manufacturing plant at which there had been complaints regarding the indoor air quality is presented. Phospholipid fatty acid characterization of the biomass enabled contamination point source determination. In comparison with samples taken outdoors, increases in the relative proportion of trans PLFA, reflecting shifts in the physiological status of viable airborne Gram-negative bacteria, were detected in the indoor air samples at a majority of sampling sites. Received 29 September 1998/ Accepted in revised form 8 January 1999     

Expression of angiotensin type 1 and 2 receptors in brain after transient middle cerebral artery occlusion in rats

Angiotensin II (Ang II) type 2 receptors (AT2Rs) have been associated with apoptosis. We hypothesized that AT2Rs are increased in stroke and may contribute effects of stroke to the brain. To test this, we have examined the expression of Ang II type 1 receptor (AT1R), AT2R and Ang II levels in the brain 24 h after transient middle cerebral artery occlusion (MCAO). The densities of AT1R and AT2R were measured by quantitative autoradiography (n=6). The levels of Ang II were measured by radioimmunoassay (RIA) (n=6) and by immunohistochemistry (n=3). AT1R levels on autoradiography showed a significant decrease (0.87±0.06 to 1.39±0.07 fmol/mg, p<0.01) in the ventral cortex of the stroke side compared to the cortices of non-stroke (NS) rats (n=4). There was no significant difference on ATIR in the contralateral verbal cortex of the stroke rats compared to NS control. In contrast, levels of AT2R in the ventral cortex of both the stroke and the contralateral sides were significantly increased (0.77±0.06, p<0.05 and 0.91±0.05, p<0.01 compared to 0.60±0.03 fmol/mg tissue, respectively). RIA showed that Ang II in the ventral cortex of both the stroke and the contralateral sides were significantly increased (241.63±47.72, p<0.01 and 165.51±42.59, p<0.05 compared to 76.80±4.10 pg/g tissue, respectively). Also, Ang II in the hypothalamus was significantly increased (179.50±17.49 to 118.50±6.65 pg/g tissue, p<0.05). Immunohistochemistry confirmed the increase of Ang II. These results demonstrate that brain Ang II and AT2Rs are increased whereas AT1Rs are decreased after transient MCAO in rats. We conclude that in stroke, Ang II and AT2R are activated and may contribute neural effects to brain ischemia.     

A comparison between Andersen (ACFM) and Reuter Centrifugal Sampler (RCS-plus) for indoor sampling of airborne molds

This paper shows first results regarding thecollection efficiency and suitability of theAndersen-6-stage cascade impactor (ACFM,Graseby Andersen) and the Reuter CentrifugalSampler (RCS-plus, Biotest Diagnostics Corp.,Denville, NJ) in measuring concentrations ofindoor airborne viable mold spores. The ACFMshowed significantly higher numbers of colonyforming units than the RCS-plus. For exposureassessment the lower sampling efficiency of theRCS-plus in comparison to the ACFM is not soimportant, as no sampler is able to record theactual concentration of airbornemicroorganisms. For fungal exposure assessmentit is more important to compare indoor andoutdoor mold spore concentration and theindoors and outdoors occuring species. The ACFMdoes not overload as quickly as the RCS-plus,so there is not so much overgrowing andinhibition of colonies and it is easier tomaintain pure cultures. The advantages of theRCS plus are the simpler handling and, the optionof changing flow-rate. Even though each sampler hasits set of advantages and disadvantages, bothcan serve a useful function in airborne fungalspore sampling.     

A novel silicon complex is as effective as sodium metasilicate in enhancing the collagen-induced inflammatory response of silicon-deprived rats

An experiment was conducted with rats to determine whether silicon deprivation affects the inflammatory response to the injection of type II collagen, and to compare the effectiveness of the organic complex arginine silicate inositol (ASI) with inorganic silicon (NaSiO₃) in mitigating any observed change in response. Dark Agouti rats were fed a ground corn–casein–safflower-based diet containing about 2.8 mg Si/kg. The experimental variables were supplemental 0 and 35 mg Si/kg as either ASI or NaSiO₃. After five weeks on their respective treatments, each rat was injected with type II collagen and euthanized four weeks later. Urine was collected before injection during week five and week nine before euthanasia. The silicon-supplemented rats generally exhibited a more marked inflammatory response than the silicon-deprived rats. The circulating number of lymphocytes was higher (p<0.003) and number of neutrophils was lower (p<0.008) in silicon-deprived than silicon-supplemented rats. ASI and NaSiO₃ were about equally effective in enhancing these changes. Post-injection of tibial release of prostaglandin E₂ (PGE₂) (p<0.04), urinary excretion of magnesium (p<0.03) and deoxypyridinoline (p<0.009), and plasma osteopontin (p<0.009), magnesium (p<0.0007) and copper (p<0.004) were higher in silicon-supplemented than silicon-deprived rats. The increases in plasma magnesium (Si×sex, p<0.04) and copper (Si×sex, p<0.02) were more marked in male than female rats. One but not the other silicon supplement when compared to silicon deprivation significantly affected the tibial release of PGE₂, and plasma copper and iron concentrations. However, with the exception of the pre-injection urinary excretion of helical peptide, no other of the variables determined was significantly different between rats fed ASI and those fed NaSiO₃. The findings suggest that, in rodents, physiological amounts of silicon promote the immune response, sex may influence the response to dietary silicon, and that both organic silicon complexes and inorganic silicon are similarly effective in preventing changes in inflammation induced by silicon deprivation.     

Assessment of airborne endotoxin in sandstorm dust and indoor environments using a novel passive sampling device in Al Zulfi city,Saudi Arabia – Establishing threshold exposure levels

The impact of sandstorm dust events affects local air quality and public health. These issues are becoming of greater concern in Saudi Arabia. There is a significant lack of research on airborne endotoxin exposure and analysis in the Middle East countries and no coherent body of research exists focusing on sandstorm dust in worldwide. In this study, we used a novel design of an aluminum foil plate (AFP) electrostatic dust cloth (EDC) for the passive air sampling of sandstorm dust. A total of 38 sandstorm dust samples were collected during sandstorm episodes occurring between January and April 2020 in both indoor (7 days, n = 20) and outdoor environments (24 h, n = 18). After exposure, and following an extraction procedure, bacterial endotoxin levels were measured using the Limulus Amoebocyte Lysate (LAL) gel clot method. The study highlights that the airborne endotoxin level observed was between 10 and 200 EU/m² in both indoor and outdoor environments, during a sandstorm event. Agricultural activities and farmhouses observed higher airborne endotoxin levels. In general, increased endotoxin levels were related to the severity of the sandstorms. Given that the observed values were high as per existing guidelines for respiratory health, we recommend the setting an occupational airborne exposure limit for bacterial endotoxin. This is the first report and further studies across various sandstorm-hit regions will need to be undertaken, together with various sampling methods, in order to assess for seasonal and geographic trends.     

Long-term use of high-efficiency vacuum cleaners and residential airborne fungal-spore exposure

Exposure to fungal allergens is an important contributor to allergic respiratory disease, but information on the efficacy of residential fungal allergen-avoidance in allergic-disease management is lacking. Using vacuum cleaners with high-efficiency exhaust filtration is one method recommended for reducing residential allergen exposure levels, but their use to reduce fungal-spore exposure levels has not been evaluated. To evaluate the effectiveness of high-efficiency vacuuming to control airborne fungal-spore levels, fungal bioaerosols were repeatedly assessed over the course of 10 months in homes randomly assigned to groups using either conventionally filtered (control) or high-efficiency-filtered vacuum cleaners for routine vacuum cleaning. Air samples were analyzed for three fungal-spore categories representing taxa with predominantly outdoor sources and one representing taxa that commonly have indoor sources. In a two-way analysis of variance, sampling period had a significant effect on mean levels of all fungal-spore categories. Vacuum cleaner type had a marginally significant effect on the indoor spore category, with one high-efficiency vacuum group mean (of three) significantly lower than one control mean. No effect was observed of vacuum cleaner type on outdoor spore categories. Including home-environment variables in analysis of covariance models strengthened the effect of the vacuum-type treatment on the indoor spore category, with no effect on the three outdoor spore categories. Decreased indoor spore levels vs. controls were only observed in high-efficiency vacuum groups during the last sampling period, at the end of the heating season. The results suggest that using a vacuum with high-efficiency filtered exhaust could have some modest effectiveness in controlling airborne fungal-spore exposure in homes when infiltration of outdoor air is very limited.     

The ability of membrane potential dyes and calcafluor white to distinguish between viable and non-viable bacteria

Various dyes were assessed for their ability to discriminate between viable and non-viable bacteria. Two methods of killing were employed: by heat treatment or by gramicidin treatment. Staining was carried out in two ways; by staining directly in the medium or by washing cells prior to staining in buffer. Carbocyanine and rhodamine 123 dyes only exhibited small changes in fluorescence between viable and non-viable populations of bacteria. Both oxonol dye (bis 1,3-dibutylbarbituric acid trimethine oxonol) and calcafluor white proved much more useful.     

Ten types of microscopically identifiable airborne fungal spores at Leiden,The Netherlands

A universal method for the complete assessment of atmospheric fungal spores does not exist, which is continuous, volumetric and non-selective, and offers at the same time reliable identification of the collected spores. To perform a survey of airborne fungal spores, a choice has to be made between a viable and non-viable method. For the study carried out in Leiden, the non-viable, continuous volumetric method has been employed, showing the results over a period of 10 years, for 10 microscopically identifiable fungal spore types. Of this selection,Cladosporium spores have by far the highest airborne quantities, with an average annual total of the daily averages of over 700 000.Botrytis, Ustilago andAlternaria follow with much lower spore concentrations of between 20 000 and 30 000 as annual totals. The spore types ofEpicoccum, Erysiphe, Entomophthora, Torula, Stemphylium, andPolythrincium are represented with annual sums lower than 10 000. A spore calendar shows the overall seasonal appearance of the 10 selected types.     

Comparison of dichloran rose bengal chloramphenicol and Sabouraud dextrose agar with cycloheximide and chloramphenicol for airborne mold sampling

The more the mold species isolated on a culture medium, the more the sampling environment is represented accurately. According to the sampling purpose, it is crucial to use the best culture medium for mold. However, no study is available regarding the comparison of dichloran rose bengal chloramphenicol (DRBC) and Sabouraud dextrose agar with cycloheximide and chloramphenicol (SDA-CHX-CHL) culture media in terms of their application for airborne sampling, isolation, and identification of fungi. Airborne mold samples were impacted onto both DRBC and SDA-CHX-CHL, simultaneously using single-stage Andersen sampler. The limit of detection (LOD) value for airborne mold count was 7 CFU m^?3 (1 colony growth on the Petri dish). The total mold counts (TMC) ranged between <7 and 504 CFU m^?3 (med 56 CFU m^?3) and <7 and 1218 CFU m^?3 (med 259 CFU m^?3), collected on SDA-CHX-CHL and DRBC, respectively. Significantly higher TMC were observed on DRBC than on SDA regardless of the sampling environment (i.e, indoor or outdoor) (p < 0.05). Among the most predominant mold genera, observation frequencies of Penicillium spp. and Aspergillus spp. on both culture media were found to be more than 70%. Observation frequencies of Cladosporium spp., Alternaria spp., and yeast were found to be higher in samples collected on DRBC than those on SDA-CHX-CHL. Finally, DRBC was found to be superior to SDA in terms of both number of colonies and number of genera isolated from the air.     

Brain peptide reverses effect of morphine on human lymphocytes

E-rosette formation by human lymphocytes incubated with sheep red blood cells (sRBC) is inhibited by morphine. We studied the ability of the opiate antagonists naloxone and Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH₂) to block this action. Active E-rosette formation by lymphocytes incubated with morphine was reduced from the control of 35.7±1.7% to 23.7±1.5% (p<0.001). Similarly, total E-rosette formation was reduced by morphine from the control of 65.8±1.3% to 53.2±2.9% (p<0.001). These effects were blocked by co-incubation of the lymphocytes with either Tyr-MIF-1 or naloxone (p<0.05). Tyr-MIF-1 was active (p<0.05) at concentrations as dilute as 10⁻¹³M. These results indicate that the neuropeptide Tyr-MIF-1 exerts an antiopiate effect at the human T-lymphocyte.     

Long-term use of high-efficiency vacuum cleaners and residential airborne fungal-spore exposure

Exposure to fungal allergens is an important contributor to allergic respiratory disease, but information on the efficacy of residential fungal allergen-avoidance in allergic-disease management is lacking. Using vacuum cleaners with high-efficiency exhaust filtration is one method recommended for reducing residential allergen exposure levels, but their use to reduce fungal-spore exposure levels has not been evaluated. To evaluate the effectiveness of high-efficiency vacuuming to control airborne fungal-spore levels, fungal bioaerosols were repeatedly assessed over the course of 10 months in homes randomly assigned to groups using either conventionally filtered (control) or high-efficiency-filtered vacuum cleaners for routine vacuum cleaning. Air samples were analyzed for three fungal-spore categories representing taxa with predominantly outdoor sources and one representing taxa that commonly have indoor sources. In a two-way analysis of variance, sampling period had a significant effect on mean levels of all fungal-spore categories. Vacuum cleaner type had a marginally significant effect on the indoor spore category, with one high-efficiency vacuum group mean (of three) significantly lower than one control mean. No effect was observed of vacuum cleaner type on outdoor spore categories. Including home-environment variables in analysis of covariance models strengthened the effect of the vacuum-type treatment on the indoor spore category, with no effect on the three outdoor spore categories. Decreased indoor spore levels vs. controls were only observed in high-efficiency vacuum groups during the last sampling period, at the end of the heating season. The results suggest that using a vacuum with high-efficiency filtered exhaust could have some modest effectiveness in controlling airborne fungal-spore exposure in homes when infiltration of outdoor air is very limited.     

Arginine vasotocin and aggression in rats

Pinealectomized (PX) and sham animals were administered either AVT (1 ng/kg) or vehicle (1 ml H₂O). Boxing-kicking-biting episodes were significantly reduced by administration of AVT, F(1,21)=8.09,p<0.009. Crawling under behavior was increased in PXH₂O animals (p<0.05). Non-aggressive rearing was decreased by administration of AVT, F(1,21)=4.75, p<0.04. These results are in agreement with previous findings that AVT lowers emotionality-arousal. Results are discussed in terms of vasotonergic effects on central monoamines.     

Composite Sampling Approaches for Bacillus anthracis Surrogate Extracted from Soil

Any release of anthrax spores in the U.S. would require action to decontaminate the site and restore its use and operations as rapidly as possible. The remediation activity would require environmental sampling, both initially to determine the extent of contamination (hazard mapping) and post-decon to determine that the site is free of contamination (clearance sampling). Whether the spore contamination is within a building or outdoors, collecting and analyzing what could be thousands of samples can become the factor that limits the pace of restoring operations. To address this sampling and analysis bottleneck and decrease the time needed to recover from an anthrax contamination event, this study investigates the use of composite sampling. Pooling or compositing of samples is an established technique to reduce the number of analyses required, and its use for anthrax spore sampling has recently been investigated. However, use of composite sampling in an anthrax spore remediation event will require well-documented and accepted methods. In particular, previous composite sampling studies have focused on sampling from hard surfaces; data on soil sampling are required to extend the procedure to outdoor use. Further, we must consider whether combining liquid samples, thus increasing the volume, lowers the sensitivity of detection and produces false negatives. In this study, methods to composite bacterial spore samples from soil are demonstrated. B. subtilis spore suspensions were used as a surrogate for anthrax spores. Two soils (Arizona Test Dust and sterilized potting soil) were contaminated and spore recovery with composites was shown to match individual sample performance. Results show that dilution can be overcome by concentrating bacterial spores using standard filtration methods. This study shows that composite sampling can be a viable method of pooling samples to reduce the number of analysis that must be performed during anthrax spore remediation.     

Pain, wheal and flare in human forearm skin induced by bradykinin and 5-hydroxytryptamine

Pain was induced in 19 healthy individuals by double-blind injections into the forearm skin of 0.05 ml of physiological saline with or without active substances added. Bradykinin (0.5 nmol), 5-hydroxytryptamine (0.5 nmol) and a mixture of the two substances in half dosage (0.25 nmol + 0.25 nmol) caused significantly more pain than saline (p<0.05). The three test solutions also induced wheal and flare responses significantly more pronounced than saline. Bradykinin induced significantly more pain and more wheal than 5-hydroxytryptamine (p<0.05) but a significantly smaller flare (p<0.01). A dissociation between induced pain and flare was thus demonstrated.