Detection and identification of moulds in dutch houses and non-industrial working environments

The mycoflora of indoor non-industrial environments is reported from “case” studies in The Netherlands. Both air sampling by a RCS-Reutcr centrifugal air sampler and surface sampling by swabs and cellotape preparations were carried out in homes, archives and libraries, musea, offices and schools. Common species encountered in these indoor environments are Aspergillus versicolor, Penicillium brevicompactum, P. chrysogenum, Cladosporium spp. and the xerophilic fungi Eurotium spp. and Wallemia sebi. Aspergillus fumigatus, Scopulariopsis spp. and Stachybotrys chartarum were occasionally isolated. It is not always possible to detect the mycoflora growing on surfaces by air sampling. Therefore direct microscopical examination and sampling from surfaces in addition to air sampling is strongly recommended for the detection of viable moulds in indoor environments. Selection of the most suitable media for isolation of fungi is discussed.     

Fungi associated with dried fish in Sri Lanka

The mycoflora of dried-salted fish from markets in Kandy, Sri Lanka was studied with emphasis on visibly spoiled dried fish. A total of 61 fungal isolates from 25 dried-fish were isolated and identified. The most prevalent fungus wasAspergillus niger. Species ofAspergillus flavus, A. fumigatus, A. glaucus, A. restrictus, Aureobasidium spp.Basipetospora halophila (a genuinely halophilic fungus)Cladosporium herbarum, Gliomastix, spp.,Penicillium chalybeum andPenicillium expansum were present. The isolated fungi did not grow in synthetic media containing more than 30% sodium chloride.Aureobasidium spp. andGliomastix spp. did not grow on dried-fish under laboratory conditions. The protective exoskeleton appeared to prevent fungal growth on dried shrimp. TheA. flavus strains isolated were not aflatoxigenic.     

Treatment of lentil (Lens culinaris. L) seeds with various concentrations of salts to check their efficiency against seed-borne mycoflora during storage

Seed sample of lentil collected from the Swabi district were treated with NaCl and KCl at 0.01, 0.1 and 1.0% (w/w). Seed-borne mycoflora was observed at 0, 20, 40, 60 and 80 day intervals. Seed treatment with both the salts was found to be effective against storage fungi; however, KCl was more efficient against storage mycoflora such as Aspergillus species when compared with NaCl. With the passage of time, the incidence of deep-seated fungi was observed in salt-treated seed samples while untreated seed sample showed heavy infestation by the species of Aspergillus. Seed viability also remained unaffected in storage, except in seeds heavily infested with seed-borne mycoflora. Aspergillus spp. were the main cause of seed rot. Surface sterilisation of seeds with 1% Na(OCl)₂ reduced the fungal infestation of seeds. Among various concentrations of salts, 0.1% (w/w) of both salts were better in controlling seed-borne mycoflora.     

Mycoflora and aflatoxin production in pigeon pea stored in jute sacks and iron bins

The mycoflora, moisture content and aflatoxin contamination of pigeon pea (Cajanus cajan (L.) Millisp) stored in jute sacks and iron bins were determined at monthly intervals for a year. The predominant fungi on freshly harvested seeds wereAlternaria spp.,Botryodiplodia theobromae, Fusarium spp. andPhoma spp. These fungi gradually disappeared from stored seeds with time and by 5–6 months, most were not isolated. The fungi that succeeded the initially dominant ones were mainly members of the generaAspergillus, Penicillium andRhizopus. Population of these fungi increased up to the end of one year storage. Higher incidence of mycoflora andAspergillus flavus were recorded in jute-sack samples throughout the storage period. The moisture content of stored seeds was found to fluctuate with the prevailing weather condition, being low during the dry season and slightly high during the wet season. The stored seeds were free of aflatoxins for 3 and 5 months in jute sacks and iron bins respectively. The level of aflatoxins detected in jute-sack storage system was considerably higher than that occurring in the iron bin system. Of 196 isolates ofA. flavus screened, 48% were toxigenic in liquid culture (54% from jute sacks and 41% from iron bins).     

Keratinophilic fungi on the hair of goats from the West Bank of Jordan

The mycoflora of the hair in 178 goats from the West Bank of Jordan was analysed and the frequency of occurrence and the relative importance value for the different keratinophilic fungi found were calculated. One hundred and seven species which belong to 38 genera were isolated. Thirty six of these species were either well recognised agents of mycoses (Trichophyton mentagrophytes, T. verrucosum, and M. nanum), or have been frequently isolated from human and animal lesions (Arthroderma spp., Acremonium kiliense, Alternaria alternata, Aspergillus flavus, Cladosporium carionii, and several other species). These potentially pathogenic fungal species comprised 66.9% of all keratinophilic fungi found on the hair of goats. The role of this animal as a reservoir for dermatophytes and other potentially pathogenic fungi is discussed.     

Xerophilic aflatoxigenic black tea fungi and their inhibition by Elettaria cardamomum and Syzygium aromaticum extracts

Black tea is consumed worldwide and is believed to play a role in cancer prevention. Xerophilic aflatoxigenic fungi are highly hazardous contaminants of tea since they are associated with tea quality impairment and human health risk. The present study reports isolation of such xerophilic and aflatoxigenic fungi associated with marketed tea. Twenty different tea samples collected from the local markets of Tamilnadu, India were investigated for fungal contamination. The results indicated contamination by 0.38% Aspergillus flavus. Other common contaminant fungi including Penicillium spp. (0.30%), Pacelomyces spp. (0.14%), and Mucor spp. (0.19%) were also isolated. Amongst the fungi isolated Aspergillus niger ML01 and A. flavus ML02 were found to be xerophilic aflatoxigenic mycoflora. Phylogenetic analysis based on 28S rRNA revealed their close ancestry. The chloroform and acetone extracts of spices Elettaria cardamomum and Syzygium aromaticum exhibited antifungal inhibitory activity on growth and toxin elaboration of both these xerophilic tea contaminants A. niger ML01 and A. flavus ML02. The results advocate the use of these spices plant or their extracts as novel antimicrobials which may add preservation and flavour in marketed tea.     

Studies on fungi in the root region

Summary Tabulated information on the colonization of roots of barley, cabbage and dwarf bean by fungi during the first 10 days of root development is given. These data, obtained by isolation and direct observation studies, are discussed in relation to previous observations on the association of fungi with the roots of healthy crop plants.The results indicate that initial root colonization may be by any of a wide range of soil fungi, but that this mixed population rapidly gives way to a stable and typical root-surface mycoflora (dominated by such fungi asFusarium spp.,Cylindrocarpon radicicola, Gliocladium spp., andPenicillium spp.     

Root-surface mycoflora of cassava (Manihot esculenta) and post harvest rot of the tubers

The root-surface mycoflora of cassava were isolated from roots washed in serial changes of sterile distilled water and plated out on potato-dextrose agar. A small group of fungi which included Aspergillus niger, Botryodiplodia theobromae, Fusarium solani, Penicillium javanicum, Penicillium sp., and Trichoderma sp. were found to be consistently associated with the root surface. While the isolates, B. theobromae and F. solani were found to be aggressively pathogenic on freshly harvested cassava tubers causing extensive rot, A. niger was only mildly so. The root-surface mycoflora, therefore, includes fungi which have been reported as the most important in postharvest deterioration of the tubers. The removal of the rhizoplane microflora by surface-sterilization using calcium hypochlorite or Clorox and subsequent incubation in loosely tied polyethylene bags extended the storage life of the tubers considerably.     

The mycobiota from seeds of Shrunken-2 (sh2) Sweet Corn

Sixteen species of fungi were isolated from the pericarp, endosperm, scutellum, and embryo of low (<75% germination), moderate (75–89%), and high vigor (>89%) seedlots of supersweet sweet corn (sh2) hybrids over two years. Most frequently isolated species werePenicillium oxalicum, Rhizopus arrhizus (14.5%), andRhizopus spp (17.4%).Fusarium moniliforme, a common inhabitant in field or dent corn, was isolated from only 2.4% of the samples. The low mean isolation values may be the result of poor conditions for infection or the data may reflect the methods used including sterilization techniques or random tissue selection.Aspergillus niger, F. moniliforme, andP. oxalicum, were isolated at a significantly greater level than other fungi from the high vigor hybrids at 0.89, 0.46 m and 4.46 respectively compared to 0.04, 0, and 1.82 for the low vigor hybrids. When Total Fungi were compared mean isolations were significantly greater from the high vigor hybrids at 11.96, the moderate 8.79, or low 4.86 vigor groups. When data from seed sources for all vigor groups were compared, significantly greater mean isolations were obtained from Illinois Foundation Seed hybrids forCladosporium sp,R. arrhizus, andRhizopus spp., but greater rates were obtained forFusarium oxysporum from the Asgrow hybrids. Isolation frequencies for the 16 species were not significantly different between the seed tissue types from any of the hybrids evaluated during this investigation. Results from this study showed that there is a diverse group of fungi present within thesh2 seed and seed treatments must be developed which will minimize seed rot and seedling blight from both internal seedborne and external pathogens.     

In situ seed baiting to isolate germination-enhancing fungi for an epiphytic orchid,Dendrobium aphyllum (Orchidaceae)

Orchid conservation efforts, using seeds and species-specific fungi that support seed germination, require the isolation, identification, and germination enhancement testing of symbiotic fungi. However, few studies have focused on developing such techniques for the epiphytes that constitute the majority of orchids. In this study, conducted in Xishuangbanna Tropical Botanical Garden, Yunnan, China, we used seeds of Dendrobium aphyllum, a locally endangered and medicinally valuable epiphytic orchid, to attract germination promoting fungi. Of the two fungi isolated from seed baiting, Tulasnella spp. and Trichoderma spp., Tulasnella, enhanced seed germination by 13.6 %, protocorm formation by 85.7 %, and seedling development by 45.2 % (all P?Epulorhiza, another seed germination promoting fungi isolated from Cymbidium mannii, also enhanced seed germination (6.5 %; P?P?Trichoderma suppressed seed germination by 26.4 % (P?Tulasnella was the only treatment that produced seedlings. Light increased seed imbibition, protocorm formation, and two-leaved seed development of Tulasnella inoculated seeds (P?Tulasnella be introduced for facilitating D. aphyllum seed germination at the protocorm formation stage and that light be provided for increasing germination as well as further seedling development. Our findings suggest that in situ seed baiting can be used to isolate seed germination-enhancing fungi for the development of seedling production for conservation and reintroduction efforts of epiphytic orchids such as D. aphyllum.     

Host plant resistance to grain mould in germplasm accessions of pearl millet (Pennisetum glaucum [L.] R. Br.)

Abstract

The paucity of information on the moulds in Indian pearl millet (Pennisetum glaucum) led to the studies that were conducted at ICRISAT, India to evaluate (a) 447 germplasm accessions of 32 countries for mould reaction in rainy season, (b) threshed grain mould rating (TGMS) and mycoflora on grains of each accession, and (c) mould scores in field and in vitro. Post physiological maturity evaluation showed that 16% of the accessions secured a mould rating of 2. In TGMS, 18% were mould free and 57% secured a rating of 2 on a 1 – 9 scale. Assessment of twenty representative accessions in vitro against individual and mixed conidial suspensions (1 × 10⁽⁶⁾ conidia ml^(?1)) of Fusarium moniliforme, F. pallidoroseum and Curvularia pennisetti indicated significant correlation (r = 0.97) between the overall field and in vitro scores of mixed spores inoculations. The mycoflora for TGMS in blotter test revealed that Fusarium moniliforme, F. pallidoroseum, Curvularia pennisetti, Helminthosporium spp., Alternaria spp. and Colletotrichum spp. to be the major fungi affecting pearl millet grain. It is advisable to harvest panicles at the physiological maturity stage to obtain better quality grains. A strong negative correlation between TGMS and % GS (r = 0.4601) and positive correlation between TGMS and % UGS (r = 0.4654) indicated that, the lesser the threshed grain mould rating higher the % seed germination.     

Effect of high temperature treatment on disinfestation and quality characteristics of sorghum

Sorghum grain (two varieties) was modified to different water contents (12% to 16% wet weight basis) and heated to 60°C, 70°C and 80°C for periods of 4, 8 and 12 min. Germination, seedling vigour, seedling dry matter, free fatty acid (FFA) content, fungal contamination and infestation with the insect pest Rhyzopertha dominica (F.) were all markedly affected by heat treatment. The effectiveness of the heat treatment was also influenced by the size of the sample used. Heat treatment at 60°C and 70°C resulted in germination being unaffected or stimulated while at 80°C and the higher water contents significantly reduced, when compared to untreated controls. The dry matter of seedlings, and seedling vigour was positively correlated with germination and heat treatment. Heat treatment had no effect on FFA. All stages of the insect pest, R. dominica, were destroyed at 70°C and an 8 min exposure time. However, the water content of the sorghum was critical in determining the efficacy against this pest. The percentage fungal contamination of grain was reduced from 90% to about 25% by heat treatment. However, some grain fungi, particularly Eurotium spp., Aspergillus niger and Penicillium spp. could still be isolated from sorghum grain treated at 80°C for up to 12 minutes.     

Investigation of stored wheat mycoflora,reporting the<Emphasis Type="Italic">Fusarium</Emphasis> cf.<Emphasis Type="Italic">langsethiae</Emphasis> in three provinces of Iran during 2007

Wheat is the most important cereal produced in Iran. A mycological survey was carried out for the first time, on the stored wheat samples in Tehran, East Azarbayejan and Mazandaran provinces in 2007. Exogenous and endogenous fungi, were isolated by the method of flotation with Malachite green agar (MGA 0.25) and Freeze blotter techniques respectively. In this study, 46 species belonging to 23 different genera were isolated.Cladosporium spp. (57.1–89.2%) andAlternaria spp. (82.4–100%) species were the predominant fungal species identified as endogenous mycoflora. The predominant exogenous fungi werePenicillium spp. (78.4–92.8%) andAspergillus spp. (71.4–85.7%) species.Fusarium proliferatum was the most prevalent species ofFusarium isolates.Aspergillus niger (39.4%) andAspergillus flavus (36.7%) were the predominantAspergillus species identified as exogenous mycoflora.Aspergillus flavus (26.6%) was the predominantAspergillus species identified as endogenous mycoflora. Flotation method with MGA 0.25 recommended for isolating of hyaline fungi from wheat cereals. In this study one isolate fromFusarium species was isolated on the basis of morphology and ribosomal internal transcribed spacer classified asFusarium langsethiae but on the basis of partial translation elongation factor-1alpha gene grouped withFusarium sporotrichioides. To our knowledge, this is the first report aboutF. cf.langsethiae in Iran and Asia.     

Biological control of maize seed pathogenic fungi by use of actinomycetes

The effectiveness of twoStreptomyces spp. strains to controlpathogenic fungi was studied in stored maizegrain. The treatments included seeddisinfection and inoculation withStreptomyces spp. strains previously isolatedfrom maize rhizosphere. Actinomycete inoculumconsisted of filtered suspension and totalsuspension of fermentor-producedStreptomyces spp. strains biomass. Treatmentswith Streptomyces spp. strains aloneeffectively suppressed the development ofAspergillus spp., Curvularia lunata, andDrechslera maydis and significantly(p < 0,05) reduced the incidence ofFusarium subglutinans and Cephalosporiumacremonium. Among the inoculation treatments,nondisinfested seed inoculated with filteredsuspension was the only treatment that did notsuppress the development of Penicilliumspp. Maize seed inoculation with totalsuspension of strains was the most effectivetreatment to control the incidence of seedpathogenic fungi. The development of theDiplodia maydis was only suppressed by thecombination of seed disinfection andinoculation with total suspension of strains.Although, the strain DAUFPE 11470 showed thegreatest effectiveness for controlling thefungi pathogenic to seed, root and shootdevelopment was reduced by treatment with thisstrain.The results indicate thatStreptomyces spp. strains reduce the incidenceof seed pathogenic fungi and have potential asa biological control agent. However, an efficient methodof seed treatment with the biological controlagent must be developed before it can become anagricultural practice.     

Peanut-pod mycoflora and kernel infection

Summary The mycoflora in soil clinging to dry pods of peanuts of the Spanish variety Argentine was sampled in 2 experiments by serially washing pods for increasing periods in changes of sterile water. Of the 9 principal fungi found,Aspergillus niger, A. flavus, A. terreus, Rhizopus spp. andSclerotium bataticola were present initially in relatively small numbers and decreased rapidly in subsequent dilutions. This decrease paralleled a decrease in weight of suspended material and in percentage of soil and organic particles greater than 0.016 mm in size.Penicillium funiculosum, P. rubrum, P. citrinum, andFusarium spp. were found in large numbers and increased or slowly decreased in numbers in subsequent dilutions. In some instances variations in numbers followed trends of percentages of soil and organic particles less than 0.016 mm in size.When dry pods with this known mycoflora were allowed to hydrate over a 6-day period at 26°, 32°, or 38°C, there was extensive pod penetration and kernel infection byA. niger, A. flavus, S. bataticola andRhizopus spp. but not by other fungi. The degree ofA. flavus andA. niger infection increased with increasing temperatures.Approved by the Director as Journal Series Paper No. 135.     

Principal component analysis of interrelations in stored-wheat ecosystems infested with multiple species of insects

Principal component analysis (PCA) was used to determine the effects of infestation of bulk stored wheat by multiple species of insects at 30±2°C for 60 wk. Eight 204-liter drums containing wheat at 15.5% moisture content were used as three distinct man-made ecosystems: (a) Control system (2 drums), insect-free; (b) RST system (3 drums), artificially infested with Rhyzopertha dominica (F.),Sitophilus oryzae (L.), and Tribolium castaneum (Herbst ); and (c) COT system (3 drums), infested with Cryptolestes ferrugineus (Stephens ),Oryzaephilus surinamensis (L.), and Tribolium castaneum. The variables measured tri-weekly within each system included carbon dioxide, oxygen, temperature, grain moisture, seed damage, grain weight and volume, dust weight and volume, fat acidity values (FAV) of the wheat, seed germination, microflora including Alternaria alternata(Fr.) Keissler ,Aspergillus glaucus group, Aspergillus candidusLink , and bacteria, insects and the mite Tarsonemus granariusLindquist . PCA provided multivariate synopsis of the data quantifying several important relationships among the variables monitored. Tri-weekly and cumulative 60-wk analyses of each system showed that high bacterial counts were associated with high FAV levels;Tarsonemus numbers were positively related to Aspergillus; Alternaria and seed germination were negatively related to FAV, bacteria and grain damage; and that the number of insects was related to the presence of Aspergillus and negatively related to the presence of bacteria. Seed germination and Alternaria infection often decrease rapidly presumably because of infection by fungi of the Aspergillus glaucus group. The combined action of R. dominica and Aspergillus spp. enhanced seed damage and increased grain moisture content thus promoting bacterial growth which in turn inhibited insect and mold growth. Fat acidity values increased with time unless seed damage and bacterial infection were extensive as in the RST system.     

Impact of biocontrol agents in combination with Prosopis juliflora (Swartz) DC. in controlling the root-infecting fungi of leguminous crops

Biocontrol agents, viz., Rhizobium meliloti, Pseudomonas aeruginosa, Aspergillus niger and Trichoderma harzianum, are used as seed dressing and soil is amended with Prsosopis juliflora (Swartz) DC. plant parts like stem, leaves and flower at 1% w/w for the control of root-rot fungi. All antagonists suppressed the infection of root-rot fungi viz., Fusarium spp., Rhizoctonia solani and Macrophomina phaseolina whereas the infection of R. solani and M. phaseolina was controlled when cowpea (Vigna unguiculata L.) and mungbean (Vigna radiata L.) seeds were treated with P. aeruginosa and T. harzianum and the soil was amended with P. juliflora leaves’ powder at 1% w/w. However, germination of both the crops was observed in all treatments. Growth parameters like shoot and root length, shoot and root weight, and leaf area significantly increased in all the treatments as compared to the control parameters. P. aeruginosa and T. harzianum in combination with soil amendment with P. juliflora plant parts at 1% w/w were the most effective for the control of root-rot fungi of leguminous plants.     

Studies on inhibitory activities of antibodies against phytopathogenic fungi: an immunological approach

The major problem in agriculture is that of the fungal pathogens. In this era, where biological control is at focus, and is the centre of crop protection as well as environmental protection, the synthesis of new bio bodies is of utmost need. Fungicides available in the market, though of potential, are pathogen specific and highly pollutive. An attempt was made to raise polyclonal antibodies against Aspergillus niger. Following the particular standardised immunisation schedule, regular injections and periodic tapping were carried out. The IgG purified was used to check cross-reactivity with different crop fungi like Fusarium solani, Fusarium oxysporum f. sp. cicer (FOC), Rhizoctonia bataticola, Botryodiplodia theobromae, Curvularia sp., Alternaria porri and also with Aspergillus niger by two different methods. Liquid test media and the radial growth inhibition test performed in solid media were used to check the inhibition of fungi and cross-reactivity. The results were subjected to analysis of variance (ANOVA) by using Tukey's test at the significance level of p < 0.05. The antibodies were active against all the fungi for more than 15 days except for A. niger in which from the seventh day onwards spore germination was observed. The probable role of antibodies to detect the common antigenic molecule that may be present in all the tested fungi and their role in inhibiting these fungi is discussed.     

Competition in vitro among fungi acquired from the exoskeleton of the giant Madagascar hissing-cockroach,Gromphadorhina portentosa,and its relevance to human health

Few insects have received as much public attention as the Madagascar hissing-cockroach, Gromphadorhina portentosa, coveted by children as a household pet. Recently, this insect has been linked to allergies and asthma triggered by an antagonistic external mycoflora, and some of these fungi are known to cause serious illnesses, namely Aspergillus spp., Mucor spp. and Rhizopus spp. Other cockroach species serve as carriers of medically important fungi, but G. portentosa is of special concern because it is deliberately reared in captivity. The aim of this study was to ascertain why certain types of fungi dominate the exoskeleton’s mycoflora whereas others do not. Six ascomycetes (Alternaria alternata, Aspergillus flavus, Aspergillus niger, Cladosporium cladosporioides, Penicillium glabrum, Trichoderma viride), and two zygomycetes (Mucor racemosus, Rhizopus stolonifer) initially acquired from the exoskeleton of G. portentosa, were tested for their competitive ability to inhibit one another in vitro, using the cockroach as the primary substratum. R. stolonifer, M. racemosus, and mycoparasitic T. viride grew at a significantly faster rate compared to the other taxa, and growth rates correlated positively with conidia output. Slower colony growth rates were noted prior to physical contact, suggesting that the fungi engaged in chemical antagonism. Three genera emerged that best fit the profile of exploitative competitors – Rhizopus, Mucor, Trichoderma. The latter two genera completely inhibited the growth of all fungal taxa tested and also had some capacity to grow over the mycelium of another fungus. We conclude that M. racemosus and T. viride in particular, are successful agents of G. portentosa’s mycoflora because they utilize both exploitation and interference competition to their advantage. Eliminating or reducing the simple sugar load entering the cockroach colony may be one way to tilt the composition of the mycoflora to favour strains that are less medically significant. Thus, we emphasize reduction of the antagonistic fungi at the colony (food entry) level and less so on the exoskeleton level.     

Airborne fungi in the hospitals of metropolitan Delhi

Summary The fungal airspora of a large hospital in Delhi Metropolis was studied from May 1989 – April 1991, using Andersen Six Stage Volumetric Sampler and Burkard Personal Slide Sampler. Simultaneously, samples were also collected from outside the hospital to act as a control. Samplers were operated for 10 min. each time, at 10 - day intervals. Additional samples were also collected from different sections of 3 other hospitals. Some of the dominant forms encountered wereCladosporium spp.,Aspergillus flavus, Smut,Fusarium spp.,Aspergillus niger, Alternaria spp.,Penicillium citrinum, Aspergillus versicolor, andPenicillium oxalicum. Aspergillus flavus showed significantly high concentration inside hospital (n=66, x=53 CFU m^–3, p<0.05) as compared to outside air. The peak period for fungi was observed to be from June – September. The spore concentration was much lower in hospital units receiving filtered air as compared to control environment, but in naturally ventilated hospitals the concentration was similar to that of outside air.