Comparison of airborne spore concentrations and fungal allergen content

The exposure to spores causing health effects is usually assessed by determining the concentration of viable spores per cubic meter of air (CFU/m³).Since allergens might also be present in dead spores or smaller particles, the objective of this study was to investigate the correlation between the viable spores of Alternaria and Cladosporium at different indoor and outdoor sites and the corresponding allergen concentration detected with a specially developed ELISA (Enzyme Linked Immunosorbent Assay). In outdoor air, the results show a strong correlation between the different sampling techniques applied for viable spores (Slit-Sampler and Multistage Liquid Impinger) and between the viable spores and the allergen concentrations detected in the liquid samples of the impingers. Indoors, the number of viable spores and the allergen concentration do not correlate and the allergen load is underestimated if colony counting methods are used. This revised version was published online in June 2006 with corrections to the Cover Date.     

Grifola frondosa extract and ergosterol reduce allergic reactions in an allergy mouse model by suppressing the degranulation of mast cells

ABSTRACT

The increasing number of patients suffering from allergic diseases is a global health problem. Grifola frondosa is an edible mushroom consumed as a health food in Asia, and has recently been reported to have anti-allergic effects. We previously reported that G. frondosa extract (GFE) and its active components, ergosterol and its derivatives, inhibited the antigen-induced activation of RBL-2H3 cells. Here, we demonstrated that GFE and ergosterol also had an inhibitory effect on the degranulation of bone marrow–derived mast cells (BMMCs) and alleviated anaphylactic cutaneous responses in mice. Using an air pouch-type allergic inflammation mouse model, we confirmed that oral administration of GFE and ergosterol suppressed the degranulation of mast cells in vivo. Our findings suggest that G. frondosa, including ergosterol as its active component, reduces type I allergic reactions by suppressing mast cell degranulation in mice, and might be a novel functional food that prevents allergic diseases.     

Penicillium andAspergillus species in the habitats of allergy patients in the Tulsa, Oklahoma area

Penicillium andAspergillus have been recognized as important aeroallergens for more than 30 years, and are especially significant in indoor environments. There are over 400 species ofPenicillium andAspergillus combined, but there is little information on which species occur most frequently in the environment, or if each exhibits unique allergenic properties. A preliminary study showed no overlap between those species isolated from an outdoor site in Tulsa, Oklahoma and the species used in immunotherapy at allergy clinics in the Tulsa area. Pursuing this line of research, air samples were collected as three seasonal samples (over a 6 month period) in the homes or offices of ten allergy patients known to be allergic toPenicillium and/orAspergillus. Twenty three species ofPenicillium and 12 species ofAspergillus were identified from these samples through isolation, macroscopic, and microscopic examination.Penicillium corylophilum, P. glabrum, Aspergillus niger, andA. flavipes were the most abundant species isolated, supporting the data obtained in a preliminary study. At least in the Tulsa area, it appears that atopic patients are being tested and treated with extracts ofPenicillium andAspergillus species that are either not present or not abundant in the local indoor or outdoor environments. Additional research is necessary to determine if the environmental isolates share allergens with those species used in immunotherapy.     

Indoor air quality in schools: exposure to fungal allergens

This study examined indoor air quality within schools in Kansas City, Spokane, Santa Fe, and Orlando. Air sampling was undertaken with both Andersen Single Stage Samplers and Burkard Personal Air Samplers. The data show a wide range of indoor exposures ranging from less than 100 colony forming units (CFU/m³) for viable fungi and 100 spores/m³ for total spores in Spokane and Santa Fe to concentrations over 6000 CFU/m³ for viable fungi and 15 000 spores/m³ for total fungi in Orlando and Kansas City, respectively. In the majority of sites the indoor airspora reflected the outdoor taxa withCladosporium the most abundant genus identified; however, several indoor locations had elevated levels ofPenicillium andAspergillus indicating possible sources of indoor contamination. Airborne basidiospores and smut spores were also fairly abundant in the schools and were among the top five taxa identified. The data also indicated that the airborne concentrations vary significantly during the day and between classrooms within each school. Continued studies in schools are needed to fully assess both the exposure levels and the clinical significance to atopic children allergic to these spores.     

Mould biodiversity in homes I. Air and surface analysis of 130 dwellings

Summary Air and surface sampling to quantify and identify fungi were conducted over a 10-year period in 130 Belgian homes, including flats and houses. Homes were selected based on the medical files of allergic, mostly asthmatic patients to assess possible mould responsibility in the aetiology of the disease. Air sampling was done with Reuter centrifugal air sampler using different sampling times and incubation temperatures to detect mesophilic and thermotolerant or thermophilic fungi. More than 50 genera were detected, withCladosporium, Penicillium andAspergillus being the most common. The main species detected wereAspergillus versicolor, several species ofPenicillium, Cladosporium sphaerospermum andCladosporium herbarum. Surface sampling was conducted concomitant with air sampling. Surfaces with and without visible mould growth were tested equally by Rodac contact plates or swabs. AlthoughCladosporium herbarum was shown to be a common species in indoor environments by air sampling, it was not detected by surface sampling from visible mould growth. This finding suggests that its presence in dwellings is from an outdoor origin and not indicative of indoor proliferation.Aspergillus versicolor, Cladosporium sphaerospermun, andPenicillium chrysogenum were the most common species isolated from surfaces showing visible mould growth.     

Fungi in the indoor environment of flour mill in Lucknow

Forty species of fungi were recovered from the indoor air of flour mills by exposing petri plates containing potato dextrose, czapek-dox, and sabouraud agar media for 5 minutes. A rotorod sampler was used. The fungal flora of the nearby outdoor environment was also studied for comparison. Species of genus Aspergillus dominated in the mills, being represented by 16 species including one ascosporic species. Other species were of genera Cladosporium and Fusarium. Variations in the fungal population in different months of the survey year were also observed.

Identifiable fungal spores recovered using the rotorod sampler were Alternaria (17% occurrence), Curvularia lunata (10.6%), C. letramera (10%), Cladosporium (19%), Drechslera sp. (9%), Epicoccum sp. (5%), Pithomyces sp. (3%), Nigrospora sp. (10.5%), Stemphylium sp. (4.5%) and Torula sp. (4.5%). Mycelial fragments and unidentifiable spores were also seen in abundance.

Varying allergic responses of patients tested intradermally for the antigens of six Aspergilli, viz., Aspergillus flavus, A. fumigatus, A. melleus, A. niger, A. niveus, and A. terreus were also recorded.     

Status of airborne spores and pollen in a coir factory in Kerala,India

The prevalence of airborne fungal spores and pollen grains in the indoor and outdoor environments of a coir factory in Thiruvananthapuram district of Kerala state, India was studied using the Burkard Personal Sampler and the Andersen 2-stage Sampler for 2 years (September 1997 to August 1999). The concentration of pollen grains was remarkably lower than that of fungal spores (ratio of 1:28). There was no large difference in the concentrations and types of fungal spores between the indoor and outdoor environments, with 26 spore types found to be present indoors and 27 types outdoors; of these, 22 were common to both the environments. Aspergillus/Penicillium, Cladosporium, ‘other basidiospores’ and ascospores were the dominant spore types. The total spore concentration was highest in February and lowest in September, and it was significantly higher in 1998–1999 than in 1997–1998. Twenty viable colony-forming types were isolated from inside the coir factory. The most dominant viable fungi isolated were Penicillium citrinum, Aspergillus flavus and Aspergillus niger. The total pollen concentration was higher in the outdoor environment of the coir factory than indoors, with 15 and 17 pollen types, respectively. Grass and Cocos nucifera pollen types were dominant. The dominant spore and pollen types trapped in the two environments of the coir factory are reportedly allergenic and, consequently, workers are at risk of catching respiratory/allergic diseases.     

Pollen and Mould Allergy in Southern Sardinia (Italy): Comparison of Skin-Test Frequencies and Air Sampling Data

A study was carried out to investigate the influence of atmospheric pollen and fungi in determining allergic diseases by comparing the frequency of skin reactions to air sampling data over a 6-year period.

48% of our population reacted to at least one of the pollen and fungal extracts used. Among pollen, Gramineae gave the most frequent positive reactions, followed by Parietaria, Olea and Compositae. The most common positive skin tests in fungus sensitive patients were by extracts of Alternaria, Cladosporium, Aspergillus and Candida

As for the aerobiological survey, the general trend of pollen and molds was similar during the sampling period. The annual pollen catch did not show remarkable differences during the years sampled, whereas the total fungal spore count was highest in 1988 and 1990.

A comparison between aerobiological and clinical data revealed a good degree of concordance between total pollen counts and positive skin test frequencies for Urticaceae, Gramineae and Oleaceae but not for Compositae (high positive skin reactions and very low counts) and Cupressaceae (high counts and few skin reactions).

A less marked correlation has been found between fungal spore counts and positive skin-test frequencies as compared to pollen. Spores such as Cladosporium, which are present in large number in the air, appear to be less sensitizing, while certain spore types (e.g. Alternaria), seem to be able to sensitize patients in spite of their low atmospheric concentrations.     

Production of Trichoderma micropropagules as a biocontrol agent in static liquid culture conditions by using an integrated bioreactor system

ABSTRACT

In this study, we optimised the conditions for the production of micropropagules of Trichoderma harzianum EGE-K38 in static liquid culture in Modified Czapec Medium (MCM) containing 8?g/L glucose in an integrated tray bioreactor system designed by our research group. Incubation temperature, air flow rate, inoculum spore concentration, inoculation size, medium volume and the use of spores or agar plugs containing mycelia as inoculum were individually studied as one factor at a time. The maximum micropropagule count was 5.2?±?0.2?×?10^9?cfu/mL and dry cell weight was 17?±?2?g/L. For the subsequent drying processes, the maximum drying yield percentage ((viable micropropagule counts after drying/viable cells before drying)*100) after drying of micropropagules was 23.30% (cfu/cfu). Results obtained from our integrated tray bioreactor system showed that static liquid culture fermentation offers potential for industrial scale fungal BCAs production.     

A report on Penicillium in the intramural and extramural air of residential areas of Nagpur city (India)

Prevalence of different species of Penicillium and their concentrations per cubic meter of air were evaluated with the use of Hi-Air sampler system Mark II (Hi-Media Laboratories Ltd., India) in the air of homes (bed-rooms) at four different sites in Nagpur. At each of these sites, air sampling was done fortnightly in triplicate for 2 years duration from June 2000 to May 2002. The sampling was also done in triplicate for the outdoor air in the vicinity of each home on the same day immediately after the indoor sampling was over. The mean concentration of Penicillium colony forming units at four different sites in the indoor air was 32, 46.9, 35 and 35.4 CFU/m³, respectively, whereas in the outdoor air at these same four sites, the mean concentration was 24, 28, 25 and 25.8 CFU/m³ respectively. The Penicillium concentration in the indoor air was found to be higher in winter than in other seasons (ANOVA, p < 0.05). Concentration of Penicillium spp. in intramural environment was always higher than that in extramural environment. Statistically significant difference existed between intramural and extramural environments at all the sites, with maximum difference at a site, which is old crowded area of the city. During the 2-years investigations, 11 species of Penicillium were isolated from the indoor air while nine species were isolated from the air outside the homes. The dominant species of Penicillium in indoor as well as outdoor air were P. citrinum (33.78 and 32.81), P. oxalicum (19.70 and 22.60), and P. chrysogenum (17.64 and 14.50). The percentage of the Penicillium in the indoor air was 10.70 while it was 8.36 in outdoor air. Indoor air showed the presence of P. glaber and P. sclerotiorum, which were absent in the outdoor air.     

The Ecology of Pneumocystis: Perspectives,Personal Recollections,and Future Research Opportunities

I am honored to receive the second Lifetime Achievement Award by International Workshops on Opportunistic Protists and to give this lecture. My research involves Pneumocystis, an opportunistic pulmonary fungus that is a major cause of pneumonia (“PcP”) in the immunocompromised host. I decided to focus on Pneumocystis ecology here because it has not attracted much interest. Pneumocystis infection is acquired by inhalation, and the cyst stage appears to be the infective form. Several fungal lung infections, such as coccidiomycosis, are not communicable, but occur by inhaling < 5 μm spores from environmental sources (buildings, parks), and can be affected by environmental factors. PcP risk factors include environmental constituents (temperature, humidity, SO₂, CO) and outdoor activities (camping). Clusters of PcP have occurred, but no environmental source has been found. Pneumocystis is communicable and outbreaks of PcP, especially in renal transplant patients, are an ongoing problem. Recent evidence suggests that most viable Pneumocystis organisms detected in the air are confined to a patient's room. Further efforts are needed to define the risk of Pneumocystis transmission in health care facilities; to develop more robust preventive measures; and to characterize the effects of climatological and air pollutant factors on Pneumocystis transmission in animal models similar to those used for respiratory viruses.     

Methods for enhancing cyanobacterial stress tolerance to enable improved production of biofuels and industrially relevant chemicals

Cyanobacteria are photosynthetic prokaryotes that can fix atmospheric CO₂ and can be engineered to produce industrially important compounds such as alcohols, free fatty acids, alkanes used in next-generation biofuels, and commodity chemicals such as ethylene or farnesene. They can be easily genetically manipulated, have minimal nutrient requirements, and are quite tolerant to abiotic stress making them an appealing alternative to other biofuel-producing microbes which require additional carbon sources and plants which compete with food crops for arable land. Many of the compounds produced in cyanobacteria are toxic as titers increase which can slow growth, reduce production, and decrease overall biomass. Additionally, many factors associated with outdoor culturing of cyanobacteria such as UV exposure and fluctuations in temperature can also limit the production potential of cyanobacteria. For cyanobacteria to be utilized successfully as biofactories, tolerance to these stressors must be increased and ameliorating stress responses must be enhanced. Genetic manipulation, directed evolution, and supplementation of culture media with antioxidants are all viable strategies for designing more robust cyanobacterial strains that have the potential to meet industrial production goals.

     

Pollen grain viability in Parietaria judaica L. during the long blooming period and correlation with meteorological conditions and allergic diseases

Abstract

Pollen grain viability and starch presence in pollen were followed during the long blooming period (May-November) of Parietaria judaica, the most widespread pellitory in Italy, responsible for many cases of allergic diseases. Observations were carried out near Siena (Tuscany), in the years 1978 and 1979.

Viability is high in late spring and early autumn, and pollen grains are starchy. The presence of starchless grains is always related to low viability: a production of pollen with a low viability occurs at the beginning and end of blooming, and also in summer, during drought periods. Pollen grain viability varies widely during the blooming period, and its variations are correlated to meteorological conditions, mainly rainfall and temperature.

Allergic diseases due to pellitory, however, are mainly reported between April and July, and on the other hand a small amount of patients states that they suffer all the year round. The discrepancies between the periods of viable pollen production and of declared symptoms by patients, are discussed.     

Estimating the probability of bacterial infection using a novel biomarker among pediatric patients in the emergency department

Context: IL-27 is a novel biomarker to identify bacterial infection in children.

Objective: IL-27 was evaluated among pediatric emergency department (ED) patients and compared with procalcitonin (PCT).

Methods and results: Children undergoing blood, urine, or cerebrospinal fluid cultures had IL-27 and PCT assays performed. Bacterial infection was defined as a positive culture or a clinical diagnosis based on chart review. IL-27 and PCT were increased among patients with bacterial infection and demonstrated comparable AUC’s (0.62 versus 0.61). A decision tree incorporating IL-27, PCT, and white blood cell count improved the AUC (0.80).

Conclusion: IL-27 is a viable candidate biomarker to identify bacterial infection among ED patients and is comparable with PCT.     

A report on <Emphasis Type="Italic">Penicillium</Emphasis> in the intramural and extramural air of residential areas of Nagpur city (India)

Prevalence of different species of Penicillium and their concentrations per cubic meter of air were evaluated with the use of Hi-Air sampler system Mark II (Hi-Media Laboratories Ltd., India) in the air of homes (bed-rooms) at four different sites in Nagpur. At each of these sites, air sampling was done fortnightly in triplicate for 2 years duration from June 2000 to May 2002. The sampling was also done in triplicate for the outdoor air in the vicinity of each home on the same day immediately after the indoor sampling was over. The mean concentration of Penicillium colony forming units at four different sites in the indoor air was 32, 46.9, 35 and 35.4 CFU/m³, respectively, whereas in the outdoor air at these same four sites, the mean concentration was 24, 28, 25 and 25.8 CFU/m³ respectively. The Penicillium concentration in the indoor air was found to be higher in winter than in other seasons (ANOVA, p < 0.05). Concentration of Penicillium spp. in intramural environment was always higher than that in extramural environment. Statistically significant difference existed between intramural and extramural environments at all the sites, with maximum difference at a site, which is old crowded area of the city. During the 2-years investigations, 11 species of Penicillium were isolated from the indoor air while nine species were isolated from the air outside the homes. The dominant species of Penicillium in indoor as well as outdoor air were P. citrinum (33.78 and 32.81), P. oxalicum (19.70 and 22.60), and P. chrysogenum (17.64 and 14.50). The percentage of the Penicillium in the indoor air was 10.70 while it was 8.36 in outdoor air. Indoor air showed the presence of P. glaber and P. sclerotiorum, which were absent in the outdoor air.     

Increased heterocyst frequency by patN disruption in Anabaena leads to enhanced photobiological hydrogen production at high light intensity and high cell density

The effects of increasing the heterocyst-to-vegetative cell ratio on the nitrogenase-based photobiological hydrogen production by the filamentous heterocyst-forming cyanobacterium Anabaena sp. PCC 7120 were studied. Using the uptake hydrogenase-disrupted mutant (ΔHup) as the parent, a deletion-insertion mutant (PN1) was created in patN, known to be involved in heterocyst pattern formation and leading to multiple singular heterocysts (MSH) in Nostoc punctiforme strain ATCC 29133. The PN1 strain showed heterocyst differentiation but failed to grow in medium free of combined-nitrogen; however, a spontaneous mutant (PN22) was obtained on prolonged incubation of PN1 liquid cultures and was able to grow robustly on N₂. The disruption of patN was confirmed in both PN1 and PN22 by PCR and whole genome resequencing. Under combined-nitrogen limitation, the percentage of heterocysts to total cells in the PN22 filaments was 13–15 and 16–18% under air and 1% CO₂-enriched air, respectively, in contrast to the parent ΔHup which formed 6.5–11 and 9.7–13% heterocysts in these conditions. The PN22 strain exhibited a MSH phenotype, normal diazotrophic growth, and higher H₂ productivity at high cell concentrations, and was less susceptible to photoinhibition by strong light than the parent ΔHup strain, resulting in greater light energy utilization efficiency in H₂ production on a per unit area basis under high light conditions. The increase in MSH frequency shown here appears to be a viable strategy for enhancing H₂ productivity by outdoor cultures of cyanobacteria in high-light environments.

     

Putative Microbial Population Shifts Attributable to Nasal Administration of Streptococcus salivarius 24SMBc and Streptococcus oralis 89a

Changes in bacterial composition of nasal microbiota may alter the host’s susceptibility to several infectious and allergic diseases such as chronic rhinosinusitis and allergic rhinitis. The aim of this study was to evaluate the effects of 1-week administration of a probiotic product, composed by a combination of Streptococcus salivarius 24SMBc and Streptococcus oralis 89a, on the nostril microbiota. Differences in the nasal microbiota composition were investigated by using a next-generation sequencing approach. A strong and significant decrease in Staphylococcus aureus abundance was detected immediately after the bacterial administration. Moreover, comparing the microbial networks of nostril microbiota before and 1 month after the end of treatment, we detected an increase in the total number of both bacterial nodes and microbial correlations, with particular regard to the beneficial ones. Furthermore, a less abundance of microbial genera commonly associated to potential harmful bacteria has been observed. These results suggest a potential ability of S. salivarius 24SMBc and S. oralis 89a to regulate and reorganize the nasal microbiota composition, possibly favoring those microorganisms that may be able to limit the overgrowth of potential pathogens.

     

Clinical significance of dust mite allergens

Dust mites are an important cause of allergic diseases worldwide. The traces of Dermatophagoides farinae and Dermatophagoides pteronyssinus can be found all over the world, while Blomia tropicalis are more common allergenic mites in tropical areas. A variety of allergenic components in dust mites have been found, apart from the different positive rates of IgE reactions in dust mite allergic patients, their biological characteristics, effects on the innate immune system, and especially their distribution characteristics in patients are different. Studying the relationship between dust mite allergens and clinical significance will help for diagnosis of patients and formulation of corresponding Allergen Specific Immunotherapy.

     

On the connection between the occurrence of airborne microfungi and allergy symptoms

The clinical value of investigations of the patients' environment—indoor as well as outdoor—with the purpose of showing the connection between occurrence of microfungi and allergy symptoms is demonstrated. Type I and Type III allergic symptoms were correlated to Alternaria, Botrytis, Penicillium, Cladosporium and Paecilomyces.     

An assessment of the air quality in indoor and outdoor air with reference to fungal spores and pollen grains in four working environments in Kerala, India

Qualitative and quantitative analyses of airborne fungal spores and pollen grains in four working environments (market, saw mill, poultry and cow sheds) in Thiruvananthapuram, the capital city of Kerala, India, were carried out for 2 years using Burkard Personal Slide Sampler and Andersen Two-Stage Sampler. Total spore concentration in these sites was always higher in indoor environments than in outdoor environments. Difference in concentration was not statistically significant in any of these work places except in saw mill (t test, p < 0.05). The highest spore concentration was recorded here followed by market, poultry and cow sheds. A total of 32 fungal spore types from indoor environments and 33 spore types from outdoor environments were recorded. Of them, 16 spore types were common to all the sites. Ameropsores, Cladosporium, other basidiospores, Ganoderma and Nigrospora were the dominant spore types in both indoor and outdoor environments. A total of 27 species of viable fungi from indoor and 24 species from outdoor environments were identified. Penicillium citrinum, Aspergillus flavus and Aspergillus niger were the most dominant viable fungi isolated. In contrast, total pollen concentration was always higher in outdoor environments than in indoor environments. Twenty-nine pollen types from indoor and 32 pollen types from outdoor were captured during the sampling. Poaceae, Cocos, Artocarpus, Amaranthus/Chenopodium and Tridax were the common and dominant pollen types observed in all the sites. Peak spore and pollen incidence were recorded during the late rainy and dry seasons (October–February) in both indoor and outdoor environments. The study revealed high prevalence of predominantly allergenic fungal spores and pollen grains in all the four work places. Workers/visitors are at potential risk of susceptibility to respiratory/allergic disorders.