Allergic potential of someAspergilli on saw mill workers in Lucknow,India

Fifty fungal types were isolated from the indoor atmosphere of saw mills by exposing Petri plates containing Czapek-dox Agar, Potato-dextrose Agar and Sabouraud Agar media for 5 min. The fungal flora of the outdoor surroundings was also studied for comparison. Species ofAspergilli dominated in the saw mills, being represented by 16 species including one ascosporic form. Other fungi were species ofCladosporium, Alternaria, Curvularia, Penicillium, Fusarium, etc. Variations in the fungal population in different months were also observed. Fungal spores recovered using the Rotorod Sampler wereAlternaria, Curvalaria lunata, Curvularia tetramera, Cladosporium, Dreschslera sp.,Epicoccum sp.,Pithomyes sp.,Nigrospora, Stemphylium sp. andTorula sp. Mycelial fragments and unidentifiable spores were also seen in abundance. Varying allergic responses of patients were also recorded by testing intradermally, the antigens of nineAspergilli, vizAspergillus flavus, A. fumigatus, A. japonicus, A. melleus, A. nidulans, A. niger, A. niveus, A. tammarii and A. terreus.     

Seasonal variation of fungal propagules in a fruit market environment, Nagpur (India)

The concentration of airborne fungal spores in a marketenvironment was examined to provide basic information needed to evaluatethe importance of varying levels and heterogeneity. Sampling has beencarried out by rotorod sampler and exposed Petri plate method to obtainthe quantitative and qualitative estimations respectively.Aspergillus was the most frequent and predominant genusdetected. Cladosporium, Penicillium and Alternariaspores were also fairly abundant which are well known as allergenic andpathogenic. The high concentration of airborne spores was recordedduring December–January. While the maximum concentration ofAspergillus was found during summer months in the marketenvironment.     

Airborne Pollen and Fungal Spores in Florist Shops in Worcester and in Bristol, UK: A Potential Problem for Occupational Health

Persistent allergies are common in workers in florist shops but little research has been done on the reasons for this. This paper reports an investigation of occupational exposure of florists to pollen and spores in three florist shops over a 2-week period in the autumn of 2000. In each shop three sampling methods were used: Burkard continuous volumetric samplers, deposition plates and low-tac tape for surface samples, including hands of the florists and leaves of a selection of the plants. The florists kept a record of the type of work they undertook each day and of the stock amounts of flowers in the shops. The volumetric traps collected 80 pollen and spore taxa. The average concentrations recorded through the working day were generally low but short-term peak (one hour mean) concentrations of some types were found to be relatively high or very high. For a few taxa these concentrations equalled or exceeded those typical for short-term peaks in wind dispersed pollen types in the ambient air. Fungal spore concentrations of several known allergenic types were also very high for peak periods. This was most notable for Aspergillus spp. which reached extremely high concentrations in one shop, compared with typical peak concentrations in the ambient atmosphere. Low-tac tape leaf samples demonstrated that the flowers' foliage is a major source of the fungal spores. Few pollen grains or fungal spores were found on the hands.     

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.     

Allergic contact dermatitis caused by mushrooms

The first author is allergic to skin contact with mushrooms of Suillus americanus, S. granulatus, S. grevillei, S. luteus, or S. neoalbidipes. Symptoms develop between one and two days after contact and last for approximately a week, disappearing completely without treatment. Symptoms consist of reddening, swelling, and itching, at the sites of contact with pileus cuticle mucilage of all five species. Pore layer tissues (tested for S. americanus and S. luteus) also produced strong reactions, as did pileus trama (tested for S. luteus). Spores from spore prints (tested for S. americanus and S. luteus) produced no reaction. The reaction can be avoided by wearing gloves when handling allergenic species and by washing hands promptly after working with these species. Similar cases, reported from North America, Europe, and Russia, involve Agaricus, Boletus, Lactarius, Paxillus, Ramaria, and Suillus species. Several cases involve allergy to multiple species or genera. Symptom severity varies, presumably with intensity of exposure. In one case, symptoms were renewed following ingestion. Most cases demonstrate delayed allergic contact sensitivity.     

Isolation,identification and testing for allergenicity of fungi from air-conditioned indoor environments

Twelve fungi were isolated and identified from air-conditioned rooms. Of the 12, 7 were species of Aspergillus, viz. A. niger, A. oryzae, A. fumigatus, A. terreus, A. nidulans, A. versicolor and A. parasiticus. Other fungi were Penicillium citrinum, Fusarium oxysporum, Trichoderma viride, Neurospora crassa and Alternaria alternata. A. niger was present in 80% of the locations. Some of the fungi isolated in this study could be opportunistic fungal pathogens, like A. fumigatus, A. niger and Penicillium citrinum, and were found to be allergenic. Results of this study indicate that air-conditioned rooms could be reservoirs of fungi and may cause allergic problems or infections in healthy or immunocompromised individuals living in these environments.     

Effect of medicating ingredients and other additives on microflora present on swine and chicken feeds from a Manitoba mill

The occurrence of microfloral components on medicated and non-medicated swine and chicken feed pellets and crumbles, produced in a Manitoba feed mill between June 1991 and October 1992, was determined. Addition of medicates to feeds generally decreased bacterial incidence and increased that of Eurotium spp. fungi. The effect was less pronounced when copper sulphate was added to medicated swine feeds.Contribution No. 1662.     

Incidence and significance of opportunistic fungi in leukemia patients in India

Twenty-four patients with acute leukemia were investigated for the incidence of opportunistic fungi. Culture isolations of the sputum and urine samples revealed significant levels of Candida in 14 patients; Candida albicans, C. tropicalis and C. pseudotropicalis were the predominant ones isolated. Aspergillus flavus was isolated from blood in two cases and C. albicans and a black yeast from the blood of another two. Serological studies showed fungal antibodies in seven patients; precipitins against Candida were detected in five and Aspergillus in two. Both of the Aspergillus positive cases and two patients who had rising antibodies against Candida died during the course of investigation. In this study 13 of 24 patients developed oral candidiasis.     

Antifungal activity of some marine organisms from India,against food spoilage Aspergillus strains

Crude aqueous methanol extracts obtained from 31 species of various marine organisms (including flora land faunal), were screened for their antifungal activity against food poisoning strains of Aspergillus. Seventeen species exhibited mild (+ =zone of inhibition 1–2 mm) to significant (+++ =zone of inhibition 3–5 mm) activity against one or the other strain under experiment. However, extracts of 12 species were active against all the three strains. Organisms like Salicornia brachiata(obligate halophyte), Sinularia leptocladus(Soft coral), Elysia grandifolia (Mollusks),Gorgonian sp. 2 and Haliclona sp. exhibited significant (inhibition zone of 3–5 mm) antifungal activity against one or the other strains. However,extracts of A. ilicifolius, Amphiroa sp.,Poryphyra sp., Unidentified sponge, Suberites vestigium, Sinularia compressa,Sunularia sp., Sinularia maxima, Subergorgia suberosa, Echinogorgia pseudorassopo and Sabellaria cementifera were mild (inhibition zone of 1–2 mm) to moderate(inhibition zone of 2–3 mm) active against the respective strains. The growth of A. japonicuswas significantly inhibited by the extracts ofS. leptocladus (r = 0.992, p < 0.0001)and E. grandifolia (r = 0.989, p < 0.0001).This revised version was published online in October 2005 with corrections to the Cover Date.     

Mycotoxin-producing potential of fungi isolated from red kidney beans

The predominant fungi present in samples of reject and retail red kidney beans were Aspergillus glaucus, Penicillium spp. and Alternaria spp. Together with A. ochraceus, A. flavus, Fusarium spp., and Trichoderma, these isolates from the reject beans were screened for numerous mycotoxins by TLC. The most consistently produced mycotoxins were penicillic acid (from A. ochraceus and Penicillium spp.) and Alternaria toxins (tenuazonic acid and alternariol). A. glaucus strains were tested for cytotoxicity in three tissue culture cell lines with positive results.     

Monitoring airborne concentrations of sporangia ofPhytophthora infestans in relation to tomato late blight in Emilia Romagna,Italy

Late blight caused byPhytophthora infestans is one of the most important diseases of tomato crops in Emilia Romagna. In total, 7–8 chemical sprays are applied per season to control the disease without taking into account the risk of infection. An integrated warning service for the prediction of tomato late blight was set up in order to define a more rational disease control strategy. As a part of the warning service, a number of sites monitoringPh. infestans airborne sporangia was set up for a timely disease prediction in the regional tomato growing areas. The results of 3 years monitoring ofPh. infestans sporangia are presented. They suggest that concentrations of airborne sporangia rapidly increase 1–2 weeks prior the blight onset in the field.     

Proteomic analysis of phytopathogenic fungus <Emphasis Type="Italic">Botrytis cinerea</Emphasis> as a potential tool for identifying pathogenicity factors,therapeutic targets and for basic research

Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era.     

Aerobiological,biochemical and immunological studies on some of the dominant <Emphasis Type=Italic>Aspergillus</Emphasis> species of South Assam (India)

Two years atmospheric survey of air-borne Aspergillus was carried out in the environmental conditions of South Assam. The survey revealed a total of 16 different species of Aspergillus with marked seasonal and annual variations. Aspergillus fumigatus was found to be the dominant atmospheric fungal species followed by Aspergillus flavus, Aspergillus niger, etc. Among the sample extracts tested, highest quantity of soluble protein was recorded in Aspergillus fumigatus (95.0 mg/g) whereas highest quantity of soluble carbohydrate (40.8 mg/g) and free amino acid (135.0 mg/g) was recorded in the sample extract of Aspergillus niger per gram of dry weight, respectively. The highest numbers of protein polypeptide bands were detected in the sample extract of Aspergillus fumigatus followed by Aspergillus flavus and lowest in Aspergillus niger. The maximum numbers of immunoglobulin E binding protein fractions were found in Aspergillus fumigatus, followed by Aspergillus flavus, Aspergillus clavatus, etc.     

Myrothecium: A new indoor contaminant?

In the natural environmentMyrothecium species occur as soil or leaf surface saprobes or as weak plant pathogens. In addition, some species ofMyrothecium are known to produce trichothecene mycotoxins. During a previous aerobiological investigation at two Las Vegas elementary schools,Myrothecium conidia were found to be the second most abundant spore type identified indoors from Burkard personal spore trap samples. The present study was undertaken to re-examine the schools to locate the source ofMyrothecium spores and to examine the ability ofMyrothecium to grow on indoor substrates. There were no obvious signs of contamination in the classrooms; however,Myrothecium spores occurred on about 30% of the Burkard samples. Two colonies ofMyrothecium were identified from subcultures of the Andersen samples, and three colonies were identified from carpet dust samples. Culture studies showed that a strain ofMyrothecium cinctum was able to grow on various culture media as well as on various indoor substrates including paper, cardboard, wallpaper, ceiling tiles, dry wall, carpets and cotton rug. Although there was no attempt to estimate any human health risks, these investigations are believed to be the first to document abundantMyrothecium spores from indoor air samples.     

Characteristics of ribonucleic acids isolated from Botryodiplodia theobromae pycnidiospores

Nucleic acids isolated from dormant and germinated Botryodiplodia theobromae pycnidiospores contain five distinct species of RNA. They include two ribosomal species, two ribosomal-associated species and transfer RNAs. Sedimentation coefficients of 25.1S and 18S were obtained for the two ribosomal RNA species and 5.8S and 5S for the two ribosomal-associated RNA components. Molecular weights of 1.20, 0.67, 0.054 and 0.035x10⁶ daltons were obtained after formaldehyde treatment and electrophoresis on polyacrylamide gels for these same four RNAs. Methylated nucleotides were present in the transfer RNAs and large and small ribosomal RNAs; in contrast 5.8S and 5S RNAs contained few methylated nucleotides. In addition to the 5 distinct RNA species, polyadenylate-containing RNA was isolated from both dormant and germinated spores.Published with the approval of the Director as paper no. 5006, Journal Series, Nebraska Agricultural Experiment Station. The work was conducted under Nebraska Agricultural Experiment Station Project no. 21-17.     

Allelopathic potential of an annual weed,Polypogon monspeliensis,in crops in India

The question of whether annual weeds are allelopathic under natural conditions still remains to be critically answered. Investigations were carried out to understand the involvement and mode of operation of allelopathy in an annual weed, Polypogon monspeliensis. Comparative studies of soils associated with and without the weed under field conditions revealed that there was no significant difference in toxicity of the two soils, and thus the possibility of its allelopathic effect on crops grown in the same season could be ruled out. However, soil amended with weed straw had significantly higher total phenolics including higher relative concentrations of phenolic fractions that were not detected in unamended soil. Phenolic fractions significantly affected the seedling growth of radish and cluster bean. It is likely that P. monspeliensis did not interfere chemically with the crops cultivated during the same season, but interfered with the following season crop through incorporated straw. These results indicate how a monocarpic annual such as P. monspeliensis can be allelopathic under field conditions and allelopathic potential can be managed. We suggest that before detailed investigations on allelopathy are performed as per earlier recommended protocols, data on weed life cycle pattern and agricultural practices should be collected.     

A simple PCR/RFLP analysis can differentiate between Candida albicans, Aspergillus niger, and Aspergillus fumigatus

The clinical management of immunocompromised patients depends on the rapid identification of infectious agents such as fungal pathogens. The procedure described here for accomplishing this uses a sensitive polymerase chain reaction method, previously reported, combined with restriction-enzyme digestion to distinguish between Candida and Aspergillus species and to classify Aspergillus strains.     

Cloning and expression of pkaC and pkaR,the genes encoding the cAMP-dependent protein kinase of Aspergillus fumigatus

This report describes the cloning and expression of both subunits of PKA in the opportunistic fungal pathogen Aspergillus fumigatus. The predicted translation product of the regulatory subunit, pkaR, is defined as a type II regulatory subunit. The gene encoding the A. fumigatus catalytic subunit, pkaC, contains the conserved kinase and activation domains that are characteristic of PkaC proteins. Both subunit mRNAs are expressed throughout the asexual life cycle of A. fumigatus. Message levels of pkaR and pkaC are higher during co-cultivation with alveolar epithelial cells than during culture alone.This revised version was published online in October 2005 with corrections to the Cover Date.     

Inhibition of ochratoxin A production and growth of <Emphasis Type="Italic">Aspergillus</Emphasis> species by phenolic antioxidant compounds

The phenolic antioxidants, gallic acid, vanillic acid, protocatechuic acid, 4-hydroxybenzoic acid, catechin, caffeic acid, and chlorogenic acid were studied for their effects on ochratoxin A (OTA) production and fungal growth of ochratoxigenic Aspergilli. Of the 12 strains tested, which included A. alliaceus, A. lanosus, A. ochraceus, A. albertensis, A. melleus, A. sulphureus, A. carbonarius, A. elegans, and A. sclerotiorum, the greatest inhibition of OTA production was seen in A. sulphureus, A. elegans, and A. lanosus. Vanillic acid and 4-hydroxybenzoic acid were the most inhibitory to both OTA production and growth of most of the strains tested. However, A.␣ochraceus was not inhibited by either compound, and A. carbonarius was not inhibited by vanillic acid. The effect of each compound on OTA production and growth differed among strains and generally was variable, suggesting that species-specific OTA production and response to phenolic compounds may be influenced by different ecological and developmental factors. In addition, inhibition of OTA production by antioxidant compounds may be useful in determining biosynthetic and regulatory genes involved in both OTA production and stress response in ochratoxigenic Aspergilli.     

Prediction of <Emphasis Type="Italic">Alternaria</Emphasis> and <Emphasis Type="Italic">Pleospora</Emphasis> concentrations from the meteorological forecast and artificial neural network in L’Aquila,Abruzzo (Central Italy)

In a previous work a 1-year time series of fungal spore concentrations was used to calibrate an artificial neural network for the estimation of Alternaria and Pleospora concentrations associated with observed meteorological variables in the atmosphere of L’Aquila, Italy. In this article the possibility to use the neural model calibrated with observed meteorological variables to predict the future fungal spore concentration from meteorological forecast is investigated. The results show that the proposed technique appears to be a suitable device to operationally predict the Alternaria and Pleospora concentrations a few days in advance. Emphasis is given to the actual use of these predictions for establishing a preventive strategy for allergy sufferers and for an appropriate use of fungicide treatments in agricultural activities, avoiding unsafe and useless pollution of the atmosphere, crops and fields.