Relationship between indoor and outdoor airborne fungal spores, pollen, and (1→3)-β-D-glucan in homes without visible mold growth

In this exploratory study, indoor and outdoor airborne fungal spores, pollen, and (1→3)-β-D-glucan levels were determined through long-term sampling (24-h) using a Button Personal Inhalable Aerosol Sampler. The air samples were collected in five Cincinnati area homes that had no visible mold growth. The total count of fungal spores and pollen in the collected samples was conducted under the microscope and Limulus Amebocyte Lysate (LAL) chromogenic assay method was utilized for the determination of the (1→3)-β-D-glucan concentration. For the combined number concentration of fungal spores and pollen, the indoor and outdoor geometric mean values were 573 and 6,435 m⁻³, respectively, with a geometric mean of the Indoor/Outdoor (I/O) ratio of .09. The geometric means of indoor and outdoor (1→3)-β-D-glucan concentrations were .92 and 6.44 ng m⁻³, respectively, with a geometric mean of the I/O ratio equal to .14. The I/O ratio of (1→3)-β-D-glucan concentration was found to be marginally greater than that calculated based on the combined number concentration of fungal spores and pollen. This suggests that (1→3)-β-D-glucan data are affected not only by intact spores and pollen grains but also by the airborne fragments of fungi, pollen, and plant material, which are ignored by traditional enumeration methodologies. Since the (1→3)-β-D-glucan level may elucidate the total exposure to fungal spores, pollen, and fungal fragments, its I/O ratio may be used as a risk marker for mold and pollen exposure in indoor environments.     

(1 → 3)-β-<Emphasis Type="SmallCaps">d</Emphasis>-glucan in different background environments and seasons

This study was undertaken to investigate how the length of the extraction period influences the (1 → 3)-β-d-glucan (β-glucan) yield and also to examine the background concentration of β-glucan as airborne β-glucan in outdoor environments in different seasons and as concentrations in airborne and floor dust in offices. To ensure compatibility between results obtained in different laboratories, it is important to use optimal and standardised methods to extract and quantify β-glucan. In this study, an extraction period of 60 min gave the highest β-glucan yield. The median concentration of β-glucan in 44 floor dust samples was 597 μg g⁻¹ dust. The median concentration of airborne β-glucan in offices was 5.1 ng m⁻³ in the summer and 2.3 ng m⁻³ in the winter, and the outdoor median concentration in towns was 6.8 ng m⁻³. The outdoor airborne concentration of β-glucan was significantly lower in January, November and December than during the rest of year. In July, the median airborne concentration of β-glucan was 14 times higher than in January. Furthermore, the airborne concentration of β-glucan was significantly higher in July than in March, April, May, September and October. In the summertime, we found that the indoor airborne concentration of β-glucan was lower than outdoor concentrations. This is in accordance with measurements of concentrations of airborne pollen and culturable fungal spores showing higher outdoor than indoor concentrations during the summer months.     

Airborne concentrations of bacteria and fungi in Thailand border market

Thailand border market is where the local Thais, Cambodians, Laotians, and Burmeses exchange their goods and culture at the border checkpoints. It is considered to be the source of aerial disease transmission especially for foreigners because it is always very crowded with people from all walks of life. Unhealthy air quality makes this area high risk of spread of airborne diseases. This study assessed airborne concentrations of bacteria and fungi in a border market to improve exposure estimates and develop efficient control strategies to reduce health risk. The density and distribution of airborne bacteria and fungi were investigated in the Chong Chom border market in Surin Province, Thailand. Eighteen air sampling sites were taken from outdoors and various work environments including indoor footpaths, wooden handicraft shops, electronic shops, the secondhand clothing shops, and fruit market areas. Exposed Petri plate method and liquid impinger sampler were used for sampling at the breathing zone, 1.5 m above the floor level, during weekend and holiday. Meteorological factors such as relative humidity, temperature, and light intensity were collected by portable data logger. The relative humidity was 67–73%, and temperature 29–33°C, and light varied between 18 and 270 Lux m⁻². Gram-positive and Gram-negative bacteria were found at a mean value of 10⁴ CFU m⁻³, and airborne fungi of 10³ CFU m⁻³ were recorded. The highest concentration of culturable airborne microorganisms was found along the indoor footpath (9.62 × 10⁴ CFU m⁻³ and 750.00 CFU/plate/h for impingement and sedimentation methods, respectively), the fruit market area (7.86 × 10⁴ CFU m⁻³ and 592.42 CFU/plate/h for impingement and sedimentation methods, respectively), and the secondhand clothing shop (4.59 × 10³ CFU m⁻³ and 335.42 CFU/plate/h for impingement and sedimentation methods, respectively) for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The lowest concentration of Gram-positive bacteria, Gram-negative bacteria, and fungi was found only at the outdoor area at 1.53 × 10⁴ CFU m⁻³, 0.93 × 10⁴ CFU m⁻³ and 0.80 × 10³ CFU m⁻³ by means of impingement method and 136.67 CFU/plate/h, 69.25 CFU/plate/h, and 62.00 CFU/plate/h by means of sedimentation methods for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The most frequently present airborne bacteria were identified as Bacillus, Corynebacteria, Diplococcus, Micrococcus, Acinetobacter, Alcaligenes, Enterobacter, and spore former rods. Acremonium, Aspergillus, Cladosporium, Penicillium, and Sporotrichum were the most frequently found aerosol fungi genera. The distribution of airborne microorganisms correlated with relative humidity and light factors based on principal component analysis. In conclusion, the border market is a potential source of aerial disease transmission and a various hazards of bioaerosols for workers, consumers, sellers, and tourists. The bioaerosol concentration exceeded the standard of occupational exposure limit. Many major indicators of allergenic and toxigenic airborne bacteria and fungi, Acinetobacter, Enterobacter, Pseudomonas, Cladosporium, Alternaria, Aspergillus, and Penicillium, were found in the various market environments.     

A comparison of airborne and dust-borne allergens and toxins collected from home,office and outdoor environments both in New Haven,United States and Nanjing,China

In this study, the airborne and dust-borne concentrations of endotoxin, (1,3)-β-d-glucan and five house dust allergens were measured in office, home, and outdoor environments both in New Haven, United States and Nanjing, China. Air samples were collected using a BioSampler at a flow rate of 12.5 l/min for 30 min. Dust samples were simultaneously collected using a surface sampler. Dust samples went through extraction and dilution before analysis, while air samples were analyzed directly. Limulus Amoebocyte Lysate (LAL) Pyrochrome and Glucatell assays were used to quantify endotoxin and (1,3)-β-d-glucan concentration levels, respectively. Enzyme-linked sorbent assay was used to measure the dust mites, cat, dog, and cockroach allergens. The experimental results indicated that endotoxin, (1,3)-β-d-glucan and allergen concentrations vary greatly both with samples and environments. In all tested environments, endotoxin concentration ranged from 0.8 to 83.7 ng/m³ for air, and 7.8 to 14.3 ng/mg for dust. (1,3)-β-d-glucan concentration ranged from 0.1 to 9.8 ng/m³ for air, and 6.6 to 110 ng/mg for dust. Cockroach allergens were detected only in New Haven office and outdoor environments, and other allergens ranged from 0.1 to 90 ng/mg for dust samples, and from 1.5 to 1,282 ng/m³ for air samples. In general, similar profiles of allergens and toxins were observed in New Haven and Nanjing environments. Linear regression analysis showed that there were better endotoxin and (1,3)-β-d-glucan linear correlations (R ² = 0.78, 0.87, respectively) between the dust and air samples compared to those of the allergens Der f 1 and Der p 1 (R ² = 0.5, 0.7, respectively). This research contributes to the development of robust biological exposure assessment and the elaboration of airborne and dust-borne bio-mass in the living environments.     

β(1 → 3)-glucan aerosols in different occupational environments

The aim of this preliminary study was to assess exposure to β(1 → 3)-glucan as well as inhalable dust and viable fungi in different occupational environments. The study was conducted in three different industrial plants: metal plant where metalworking fluids were applied, wastewater treatment plant, and waste composting plant. In selected points simultaneously the stationary air sampling was performed to evaluate the levels of inhalable dust, β(1 → 3)-glucan, and to make a quantitative analysis of airborne fungi. All variables describing the exposure were characterized by a wide range of concentrations. The results were as follows: β(1 → 3)-glucan (1.38–65.1 ng/m³), inhalable dust (0.03–2.93 mg/m³), and fungi (0.16–285 × 10² CFU/m³). The highest concentrations for all parameters were found in the composting plant. In the composting plant, a statistically significant correlation was found between β(1 → 3)-glucan and fungal levels (r = 0.89; p < 0.05). In the metal industry and composting plant, the participation of alkali-soluble fraction was stable, exceeding 90% of all β(1 → 3)-glucan. However, in the wastewater treatment plant, its average amount was much lower—73.6%. The study showed that β(1 → 3)-glucan was present in different occupational environments and it should be taken into consideration as an important part of bioaerosols. However, more studies are required to assess the concentration levels as well as all determinants of exposure.     

Airborne dust,bacteria, actinomycetes and fungi at a flourmill

A study was carried out on suspended dust, bacterial and fungal aerosols in a four-storey flourmill building located in Giza, Egypt. Airborne microorganisms were quantitatively isolated using liquid impinger and gravimetric samplers during the period from March 2004 to February 2005. Suspended dust varied from 1.96 to 16.3 mg m⁻³ and 0.69 to 1.8 mg m⁻³ in the indoor and outdoor environments, respectively. Suspended dust was significantly greater (P < 0.05) at bran package, double roller, purifiers and flour storage units in comparison to the outdoor reference site. The dust levels exceed the occupational exposure limit (OEL) of 0.5 mg m⁻³ for flour dust. Airborne microbial counts were found at median values, between sampling locations, ranged from 0 to >10⁴ CFU m⁻³. Gram-negative bacteria were found in small numbers (0–10² CFU m⁻³). The highest concentration of actinomycetes (>10³ CFU m⁻³) was detected in the storage unit. Airborne fungal counts were found at the median values, between sampling locations, varied from 10³ to 10⁴ CFU m⁻³. The counts of airborne bacteria and fungi were significantly greater (P < 0.05) at the purifiers and double roller mill units in comparison to the outdoor reference site using the liquid impinger sampler. Microbial levels associated with bulk deposited dust averaged between 10⁵ and 10⁶ CFU g⁻¹. Alcaligenes (5.4%) Pseudomonas (3.87%) and Enterobacter (3.1%) were the predominant Gram-negative species while Bacillus (29.4%) and Micrococci (13.9%) were the major components of Gram-positive bacteria. Aspergillus and Penicillium were the predominant fungal types indoor whereas Cladosporium (35.2%) and Aspergillus species (22.2%) were the predominant fungal types outdoor. A number of allergenic and toxigenic bioaerosols were found in the flourmill workplace.     

Assessment of bioaerosols at a concentrated dairy operation

Increased bioaerosol loadings in downwind plumes from concentrated animal feeding operations (CAFOs) may increase the risk for allergy and infection in humans. In this study, we monitored airborne concentrations of culturable bacteria and fungi at upwind (background) and downwind sites at a 10,000 milking cow dairy over the course of a year. The average bacterial concentrations at the upwind site were 8.4 × 10³ colony forming units (CFU) m⁻³ and increased to 9.9 × 10⁵ CFU m⁻³ at the downwind edge of the cattle lots, decreasing to 6.3 × 10⁴ CFU m⁻³ 200 m farther downwind. At the same sites, the average fungal concentrations were 515, 945, and 1,010 CFU m⁻³, respectively. Significant correlations between the ambient weather conditions and airborne fungal and bacterial concentrations were identified. Sequence analysis of PCR-amplified DNA from bacterial clones and fungal isolates revealed genus and species level differences between upwind and downwind sites. Although we could not cultivate gram-negative bacteria, bacterial clones at downwind sites identified as being gram-negative matched with the following genera: Acinetobacter, Bradyrhizobium, Escherichia, Idiomarina, Methylobacterium, Ralstonia, and Novosphingobium. Fungal isolates from downwind matched with the following genera: Acremonium, Alternaria, Ascomycte, Aspergillus, Basidiomycete, Cladosporium, Davidiella, Doratomyces, Emericella, Lewia, Onygenales, Penicillium, Rhizopus, and Ulocladium. None of the bacterial and fungal sequence matches were affiliated with genera and species known to be pathogenic to humans. Overall, the data suggest that exposure to bioaerosols in the downwind environment decreases with increasing distance from the open-lot dairy.     

Critical roles of Asp270 and Trp273 in the α-repeat of the carbohydrate-binding module of endo-1,3-β-glucanase for laminarin-binding avidity

A carbohydrate-binding module from family 13 (CBM13), appended to the catalytic domain of endo-1,3-β-glucanase from Cellulosimicrobium cellulans, was overexpressed in E. coli, and its interactions with β-glucans, laminarin and laminarioligosaccharides, were analyzed using surface plasmon resonance biosensor and isothermal titration calorimetry. The association constants for laminarin and laminarioligosaccharides were determined to be approximately 10⁶ M⁻¹ and 10⁴ M⁻¹, respectively, indicating that 2 or 3 binding sites in the α-, β-, and γ-repeats of CBM13 are involved in laminarin binding in a cooperative manner. The binding avidity is approximately 2-orders higher than the monovalent binding affinity. Mutational analysis of the conserved Asp residues in the respective repeats showed that the α-repeat primarily contributes to β-glucan binding. A Trp residue is predicted to be exposed to the solvent only in the α-repeat and would contribute to β-glucan binding. The α-repeat bound β-glucan with an affinity of approximately 10⁴ M⁻¹, and the other repeats additionally bound laminarin, resulting in the increased binding avidity. This binding is unique compared to the recognition mode of another CBM13 from Streptomyces lividans xylanase.     

Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis

The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and β-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or β-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores.     

A comparative study on the airborne fungi in Athens,Greece, by viable and non-viable sampling methods

Airborne fungi were studied in the city of Athens using two complementary methods in which 136 concurrent samplings were carried out during the 12-month period from January until December 1998. A portable Burkard air sampler for agar plates was used for trapping the culturable portion of the mycobiota. Nineteen genera of fungi were identified and assessed in terms of total numbers and fluctuations in concentration (Alternaria, Arthrinium, Aspergillus, Aureobasidium, Botrytis, Chrysonilia, Cladosporium, Drechslera, Epicoccum, Fusarium, Mucor, Nigrospora, Paecilomyces, Penicillium, Rhizopus, Sclerotinia, Scopulariopsis, Trichoderma and Ulocladium), with the exception of those included in the Sphaeropsidales, the yeasts, and the non-sporulating fungi, which were counted as groups. A volumetric Burkard air sampler for glass slides was operating simultaneously for detecting the total mycobiota, including the non-culturable and the non-viable portion. Ascospores, basidiospores, spores of Myxomycetes, Ustilaginales, Uredinales and Erysiphales, teliospores of Puccinia, as well as conidia of the genera Curvularia, Helminthosporium, Periconia, Pestalotiopsis, Pithomyces, Polythrincium, Stachybotrys, Stemphylium and Torula were also recorded. Only seven of the genera were recovered by both samplers. The total numbers of fungal spores, which had a maximum concentration of 3,175 spores/m³, as well as the spore concentrations of the genera Cladosporium (2,565 spores/m³) and Alternaria (280 spores/m³) were underestimated by the viable method (2,435 CFU/m³ for the total, 2,169 CFU/m³ for Cladosporium and 180 CFU/m³ for Alternaria). The non-viable method fails to resolve the identification of the genera Penicillium and Aspergillus, which are major components of the airborne mycobiota (1,068 CFU/m³ and 204 CFU/m³, respectively) based on recovery by the viable method.     

Quantification of airborne Aspergillus fumigatus spores in rabbit houses by real-time polymerase chain reaction

To accurately quantify airborne Aspergillus fumigatus (A. fumigatus) spores in rabbit houses, the real-time polymerase chain reaction (real-time PCR) and culture-based counting method (CCM) were employed to determine the airborne A. fumigatus spore concentrations. The results showed that, of the three rabbit houses (A, B, and C), the average concentrations of airborne A. fumigatus spores determined by real-time PCR were 3.0 × 10³, 3.3 × 10³, and 1.5 × 10³ spores/m³ air, respectively, while those determined by CCM were 2.5 × 10², 2.8 × 10², and 1.1 × 10² colony-forming unit/m³ air (CFU/m³ air), respectively, i.e., the former concentration was 12–14 times higher than the latter one. Therefore, the conventional CCM underestimated the concentrations of airborne fungal spores, and it is insufficient to determine the microbial aerosol concentration and evaluate the health risk only using CCM.     

Structural Characterization of <Emphasis Type="Italic">β</Emphasis>-glucans of <Emphasis Type="Italic">Agaricus brasiliensis</Emphasis> in Different Stages of Fruiting Body Maturity and their Use in Nutraceutical Products

β-Glucans of Agaricus brasiliensis fruiting bodies in different stages of maturity were isolated and characterized by FTIR and NMR. These fractions had greater amount of (1→6)-β-glucan and the (1→3)-β-glucan increased with fruiting bodies maturation. Yields of β-glucans increased from 42 mg β-glucans g⁻¹ fruiting bodies (dry wt) in immature stage to 43 mg g⁻¹ in mature stage with immature spores, and decreased to 40 mg g⁻¹ in mature stage with spore maturation. Mature fruiting bodies, which included these glucans, have potential therapeutical benefits for use in nutraceutical products.     

Airborne fungi associated with ornamental plant propagation in greenhouses

The objective was to determine potential exposure to airborne fungi in greenhouses and to characterize the temporal patterns of airborne fungi in relation to environmental conditions. We analyzed air samples collected in two greenhouses. Results showed that the top 5 fungi in greenhouse 1 were Trichoderma, hyphal fragments, Aspergillus/Penicillium-like, Cladosporium, and Botrytis in a descending order. Those in greenhouse 2 were Aspergillus/Penicillium-like, Cladosporium, Botrytis, yeast-like, and hyphal fragments. Maximum concentrations of Trichoderma and total spores in greenhouse 1 were 36,426 and 49,729 spores/m³, respectively. Maximum concentrations of Aspergillus/Penicillium and total spores in greenhouse 2 were 46,961 and 71,037 spores/m³, respectively. Airborne fungal populations fluctuated dramatically within 2 h during work hours, tenfold for Aspergillus/Penicillium, 66-fold for Trichoderma, and sevenfold for total spores. QPCR detected Trichoderma harzianum ranging from 7 to 3,500 conidia E/m³. Aspergillus/Penicillium and Botrytis showed diurnal patterns, but not Trichoderma. Aspergillus/Penicillium and Cladosporium were positively correlated with temperature, relative humidity, dew point, heat index, and light and negatively with air movement and air pressure. Botrytis and Trichoderma were not correlated with the environmental factors. Greenhouse workers were potentially exposed up to 71,037 spores/m³ of airborne fungi.     

Exposure to indoor fungi in different working environments: A comparative study

In this study an attempt was made to evaluate the qualitative and quantitative fungal burden (load) in five different working environments of South Assam (India) and the possible risks of indoor fungi to employees and stored products. Fungal concentrations in different working environments were studied using a Burkard personal petriplate sampler. The survey was done in five different working environments for one year. A total of 76 fungal types were recorded in the indoor air of South Assam during the survey period. The maximum fungal concentration (5,437.6 ± 145.3 CFU m⁻³ air) was recorded in the indoor air of medical wards, followed by the paper-processing industry (3,871.7 ± 93.4 CFU m⁻³ air). However the lowest concentration was observed in the indoor air of a bakery (1,796.8 ± 54.4 CFU m⁻³ air). The most dominant fungal genera were Aspergillus (34.2%) followed by Penicillium (17.8%), Geotrichum (7.0%) and the most dominant fungal species were Aspergillus fumigatus (2,650.4 CFU m⁻³ air) followed by Aspergillus flavus (1,388.2 CFU m⁻³ air), Geotrichum candidum (1,280.3 CFU m⁻³ air), Aspergillus niger (783.3 CFU m⁻³ air), and Penicillium aurantiovirens (774.0 CFU m⁻³ air). The fungal species viz., Aspergillus fumigatus, Penicillium aurantiovirens, Aspergillus flavus, Aspergillus niger, Geotrichum candidum, and Penicillium thomii, which were recorded well above threshold levels, may lead to adverse health hazards to indoor workers. Setting occupational exposure limits for indoor fungal spores as reference values is obligatory for prevention and control of adverse effects of indoor fungal exposure.     

Purification and characterization of an endoglucanase from <Emphasis Type="Italic">Aspergillus terreus</Emphasis> highly active against barley β-glucan and xyloglucan

An endoglucanase (1, 4-β-d glucan glucanohydrolase, EC 3.2.1.4) which was catalytically more active and exhibited higher affinity towards barley β-glucan, xyloglucan and lichenin as compared to carboxymethylcellulose (CMC) was purified from Aspergillus terreus strain AN₁ following ion-exchange and hydrophobic interaction chromatography and gel filtration. The purified enzyme (40-fold) that apparently lacked a cellulose-binding domain showed a specific activity of 60 μmol mg⁻¹ protein⁻¹ against CMC. The purified enzyme had a molecular weight of 78 and 80 KDa as indicated by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and gel filtration, respectively, and a pI of 3.5. The enzyme was optimally active at temperature 60°C and pH 4.0, and was stable over a broad range of pH (3.0–5.0) at 50°C. The endoglucanase activity was positively modulated in the presence of Cu²⁺, Mg²⁺, Ca²⁺, Na⁺, DTT and mercaptoethanol. Endoglucanase exhibited maximal turn over number (K _cat) and catalytic efficiency (K _cat/km) of 19.11 × 10⁵ min⁻¹ and 29.7 × 10⁵ mM⁻¹ min⁻¹ against barley β-glucan as substrate, respectively. Hydrolysis of CMC and barley β-glucan liberated cellobiose, cellotriose, cellotetraose and detectable amount of glucose. The hydrolysis of xyloglucan, however, apparently yielded positional isomers of cellobiose, cellotriose and cellotetraose as well as larger oligosaccharides.     

Protein,fatty acid,and pigment content of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> under different light regimes

The effects of irradiance and photoperiod on growth rates, chlorophyll a, β-carotene, total protein, and fatty acid content of Chlorella vulgaris were determined. The maximum growth rate (1.13 day⁻¹) was at 100 μmol photons m⁻² s⁻¹ and 16:8-h light/dark photoperiod. Chlorophyll a and β-carotene contents significantly differed under different light regimes with chlorophyll a content lower at high irradiance and longer light duration, while β-carotene showed the inverse trend. The total protein and fatty acid content also significantly differed in different light regimes; the maximum percentage of protein (46%) was at 100 μmol photons m⁻² s⁻¹ and 16:8 h photoperiod, and minimum (33%) was at 37.5 μmol photons m⁻² s⁻¹ and 8:16 h photoperiod; the total saturated fatty acids increased, while monounsaturated and polyunsaturated fatty acids decreased with increasing irradiance and light duration.     

Effects of exogenous methyl jasmonate in elicited anthocyanin-producing cell cultures of ohelo (Vaccinium phalae)

Summary Elicitation of anthocyanin-producing cells of ohelo (Vaccinium pahalae) by both biotic (purified β-glucan and chitosan) and abiotic [sodium ferric ethylenediamine di-(o-hydroxyphenylacetate) FeEDDHA, and CuSO₄] elicitors resulted in significant enhancement of anthocyanin accumulation. Anthocyanin production increased up to 1.8 and 1.5-fold over the control in the presence of abiotic elicitors (90 μM FeEDDHA and 20 μM CuSO₄, respectively), and increased 1.9 and 1.6-fold in the presence of biotic elicitors (10 mg L⁻¹ β-glucan and 100 mg L⁻¹ chitosan). Maximum anthocyanin production with the two most effective elicitors was achieved when cultures were treated on Day 3 (β-glucan) or Day 0 (FeEDDHA) after the initiation of fresh cell cultures. A concentration-dependent response was exhibited by cultures treated with exogenous methyl jasmonate (MJ). The addition of 0.5 μM MJ alone provoked a 2–3-fold increase in anthocyanin production over that of the control; however, no additive effect on anthocyanin production was observed in any treatments which combined MJ and β-glucan or FeEDDHA. Conditioning of the cells with a preculture in either MJ, β-glucan, or FeEDDHA similarly did not enhance anthocyanin production. Inoculation of cultures elicited by MJ or β-glucan with ibuprofen, a reported inhibitor of jasmonate biosynthesis, dramatically stimulated, rather than inhibited, anthocyanin production, resulting in levels of accumulation beyond any of the tested elicitor combinations. Hypotheses for the observed influence of ibuprofen in this system are discussed.     

Pollen dispersal and deposition characteristics of Abies alba, Fagus sylvatica and Pinus sylvestris, Roztocze region (SE Poland)

Pinus sylvestris L., Abies alba Mill. and Fagus sylvatica L.—the significant forest forming tree species in Europe are important for palaeoecological interpretations based on the results of pollen analysis of fossil deposits. The potential pollen loading for Pinus sylvestris, Abies alba and Fagus sylvatica was modelled using simulated and actual vegetation maps, measured fall-speed values and pollen productivity estimates from the literature. The influx of fir pollen drops sharply with distance from the pollen source due to the high fall speed and moderate pollen productivity. The vast majority of Abies alba pollen is deposited within less than 50 m of the sampling site and a major proportion within 100 m. For beech the corresponding numbers would be 300 and 1,800 m, and for pine 1,000 and 4,500 m. The observed mean pollen accumulation rate (PAR) values for Pinus and Fagus were ca. 5,800 and 1,100 grains cm⁻² year⁻¹, respectively. In the case of Abies, the mean annual PAR for the whole region is ca. 700 grains cm⁻² year⁻¹. In SE Poland the regional signal is represented by PARs of Abies alba <200 grains cm⁻² year⁻¹ and of Fagus sylvatica <500 grains cm⁻² year⁻¹. The local presence/absence threshold values for Abies alba, Fagus sylvatica and Pinus sylvestris are >1,000 grains cm⁻² year⁻¹, >2,000 grains cm⁻² year⁻¹ and >3,500 grains cm⁻² year⁻¹ respectively.     

Incidence of allergenically significant fungal aerosol in a rural bakery of West Bengal,India

The frequency of fungal spores in the air of three different sections of a rural bakery was analyzed using a Burkard personal slide sampler and Andersen two stage viable sampler. In average concentration of spores (No./m³) was 228–26770/m³ and concentration of viable colony forming units (CFU/m³) was 65-2061 CFU/m³. Dominant fungus species both culturable and nonculturable, were species of Aspergillus and Penicillium, Cladosporiumsp., Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Cladosporium cladosporioides, Penicillium citrinum and Alternaria alternata. Seasonal variations in the spore concentrations were clearly observed in case of some fungi. Total culturable mould concentration of different bakery sections sometimes exceeded the acceptable limit for a healthy indoor environment. Antigenic extracts prepared from some dominant culturable fungi showed high level of allergenicity in skin prick tests indicating that they could be responsible for allergic respiratory dysfunction of bakery workers.This revised version was published online in October 2005 with corrections to the Cover Date.     

Purification of exo-1,3-beta-glucanase, a new extracellular glucanolytic enzyme from Talaromyces emersonii

The moderately thermophilic aerobic ascomycete Talaromyces emersonii secretes, under selected growth conditions, several β-glucan hydrolases including an exo-1,3-β-glucanase. This enzyme was purified to apparent homogeneity in order to characterise its biochemical properties and investigate hydrolysis of different β-glucans, including laminaran, a 1,3-β-glucan from brown algae. The native enzyme is monomeric with a molecular mass of ~40 kDa and a pI value of 4.3, and is active over broad ranges of pH and temperature, with optimum activity observed at pH 5.4 and 65 °C. At pH 5.0, the enzyme displays strict specificity for laminaran (apparent K _m 1.66 mg mL⁻¹; V _max 7.69 IU mL⁻¹) and laminari-oligosaccharides and did not yield activity against 1,4-β-glucans, 1,3;1,4-β-glucans or 4-nitrophenyl- and methylumbelliferyl-β-d-glucopyranosides. Analysis of hydrolysis products formed during time-course hydrolysis of laminaran by high-performance anion exchange chromatography with pulsed amperometric detection revealed a strict exo mode of action, with glucose being the sole reaction product even at the initial stages of hydrolysis. The T. emersonii exo-1,3-β-glucanase was inhibited by glucono-δ-lactone (K _i 1.25 mM) but at significantly higher concentrations than typically inhibitory for exo-glycosidases such as β-glucosidase. ‘De novo’ sequence analysis of the purified enzyme suggests that it belongs to family GH5 of the glycosyl hydrolase superfamily. The results clearly show that the exo-1,3-β-glucanase is yet another novel enzyme present in the β-glucanolytic enzyme system of T. emersonii.