Ecology of pathogenic yeasts in Amazonian soil.

In an investigation of Amazonian soil as a natural reservoir for pathogenic fungi, 1,949 soil samples collected from diverse geographical and ecological settings of the Brazilian Amazon Basin were analyzed for the presence of non-keratinophilic fungi by the indirect mouse inoculation procedure and for the presence of keratinophilic fungi by the hair bait technique. All soil samples were acidic with low pH values. From 12% of the soil samples, 241 yeast and yeastlike isolates pertaining to six genera and 82 species were recovered, of which 63% were Torulopsis and 26% were Candida species. Nine fungi with known pathogenic potentials were encountered among 43% (104) of the isolates: T. glabrata, C. guilliermondii, C. albicans, C. pseudotropicalis, C. stellatoidea, C. tropicalis, Rhodotorula rubra, and Wangiella dermatitidis. The yeast flora was marked by species diversity, low frequency of each species, random geographical distribution, and an apparent lack of species clustering. The composition and distribution of the yeast flora in soil differed from those of the yeast flora harbored by bats, suggesting that the Amazonian external environment and internal bat organs act as independent natural habitats for yeasts.     

Distribution and characterisation of entomopathogenic fungi from Korean soils

We investigated the occurrence of entomopathogenic fungi in 1080 soil samples representing multiple locations and conditions in Korea. Entomopathogenic fungi were isolated from soils using a selective medium containing dodine and antibiotics. Following an initial identification based on morphology, the fungal isolates were more precisely identified by the sequence of their nuclear ribosomal RNA (rRNA) internal transcribed spacer (ITS) regions. As a result, entomopathogenic fungi were found to occur in 32% (342 isolates) of the soil samples studied. The most abundant species were Beauveria spp. (125 isolates) and Metarhizium spp. (82 isolates). Entomopathogenic fungi were more often recovered from natural mountain and riparian soils than from agricultural habitats. The pathogenicity of isolated fungi was evaluated by using wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) larvae. It was determined that 60% (207 isolates) of the isolates were pathogenic using this model. These entomopathogenic fungi may, therefore, have potential use against a variety of agricultural pests. This is the first study of the isolation and distribution of entomopathogenic fungi in representative sampling locations throughout Korea.     

Studies on the distribution of alkalophilic and alkali-tolerant soil fungi II: Fungal flora in two limestone caves in Japan

In a series of studies on the distribution of alkalophilic and alkali-tolerant fungi, soil fungi were isolated from five alkaline calcareous soil samples in two closely located limestone caves (stalactite caves) in Japan using slightly acidic and alkaline media. Some common soil fungi that can grow in alkaline conditions were obtained in high frequencies. The growth response to pH of the isolates revealed that approximately one third (30.8%) of the isolates had the optimum pH in the alkaline range. All isolates of fourAcremonium species and twoChrysosporium species grew well in alkaline conditions, of whichAcremonium sp. andChrysosporium sp. were pronounced alkalophiles. These fungi were thought to be indigenous species in this alkaline environment. The fungal flora in the Japanese alkaline soils was considerably different from the flora reported in alkaline environments in other countries.     

Association of anurans with pathogenic fungi

In a study of 450 Amazonian anurans, we isolated yeasts and yeast-like fungi from 54 animals (Bufo granulosus, B. marinus, Dendrophrynyscus sp., Hyla geographica, H. lanciformes, Ololygon rubra, Adenomera hylaedactyla, Eleutherodactylus fenestratus, Leptodactylus fuscus, L. ocellatus, L. pentadactylus). The internal organs of these animals did not show any macroscopic anomaly nor histopathology. We recovered 105 fungal isolates from the anuran liver, lung, kidney, spleen, heart and gonad. The isolates were made up of 30 fungal species, 9 of which (48 isolates, 46%) were fungi with known pathogenic potentials, namely: Candida guilliermondii, C. parapsilosis, C. tropicalis, C. glabrata, Geotrichum candidum, Aureobasidium pullulans, Wangiella dermatitidis, Trichosporon cutaneum and Exophiala werneckii. Eleven animals harbored identical fungi in more than one of their internal organs; seven animals had more than one fungal species colonizing a single organ. Our findings indicated probable natural subclinical infections of candidiasis, geotrichosis or phaeohyphomycosis, and also symbiotic presence of non-pathogenic fungi among neotropical anurans.     

Salt-tolerant phenol-degrading microorganisms isolated from Amazonian soil samples

Two phenol-degrading microorganisms were isolated from Amazonian rain forest soil samples after enrichment in the presence of phenol and a high salt concentration. The yeast Candida tropicalis and the bacterium Alcaligenes faecoalis were identified using several techniques, including staining, morphological observation and biochemical tests, fatty acid profiles and 16S/18S rRNA sequencing. Both isolates, A. faecalis and C. tropicalis, were used in phenol degradation assays, with Rhodococcus erythropolis as a reference phenol-degrading bacterium, and compared to microbial populations from wastewater samples collected from phenol-contaminated environments. C. tropicalis tolerated higher concentrations of phenol and salt (16 mM and 15%, respectively) than A. faecalis (12 mM and 5.6%). The yeast also tolerated a wider pH range (3-9) during phenol degradation than A. faecalis (pH 7-9). Phenol degradation was repressed in C. tropicalis by acetate and glucose, but not by lactate. Glucose and acetate had little effect, while lactate stimulated phenol degradation in A. faecalis. To our knowledge, these soils had never been contaminated with man-made phenolic compounds and this is the first report of phenol-degrading microorganisms from Amazonian forest soil samples. The results support the idea that natural uncontaminated environments contain sufficient genetic diversity to make them valid choices for the isolation of microorganisms useful in bioremediation.     

Studies on pathogenic dematiaceous fungi

Two hundred and twenty-six samples of woody materials, vegetable matter and soil were processed by the direct plating and mouse inoculation technique for the isolation of pathogenic dematiaceous fungi. The species of fungi isolated were Fonsecaea pedrosoi — 13, Cladosporium carrionii — 7 and Phialophora verrucosa — 4 isolates. The mouse inoculation technique was found to be much better than direct plating for the recovery of these fungi. Woody plant materials proved to be a good sample source for pathogenic dematiaceous fungi contributing about 90% of the isolates. All the isolates were pathogenic for mice as evidenced by the presence of dark nodular lesions containing fungal elements in the organs of experimentally infected animals.     

The isolation of keratinophilic fungi from African soils

Summary Twenty-seven soil samples from Kenya and 9 from Egypt were investigated for the presence of keratinophilic and pathogenic fungi. No systemic pathogenic fungi was obtained, while a total of 25 dermatophytes belonging to four species was isolated. The majority of the Kenya isolates belonged toM. gypseum, while Egyptian isolates belonged toT. mentagrophytes. Shady sites frequented by humans or animals produced higher incidences of dermatophytes in Kenya than those of the unshady isolated areas.     

Distribution,occurrence and characterization of entomopathogenic fungi in agricultural soil in the Palestinian area

The occurrence of entomopathogenic fungi was investigated in irrigated vegetable fields and citrus orchards soils, over a nine-month period (April-December 1999), using the Galleria bait method (GBM). Entomopathogenic fungi were found to occur in 33.6% of the soil samples studied, with positive samples yielding 70 fungal isolates, belonging to 20 species from 13 genera. Conidiobolus coronatus was the most frequent and abundant entomopathogenic species recovered, comprising 31.4% of the total number of isolates. Soil pH, soil moisture content and the geographical location had minor or no effect on the isolation of entomopathogenic fungi in the fields studied. On the other hand, organic matter content of soil, and vegetation type were found to significantly affect the occurrence of entomopathogenic fungi in soil habitats, with orchard fields yielding larger numbers of isolates than vegetable fields. Using Koch's postulates the pathogenicity of fungal isolates to Galleria larvae was found to range from 16-100% (mortality rate). Isolates of C. coronatus proved to be the most virulent isolates recovered. The effect of media and temperature on mycelial growth rate, conidial production and conidial germination of six entomopathogenic fungal species (C. coronatus, Entomophaga grylli, Erynia castrans, Hirsutella jonesii, Paecilomyces farinosus and Sporodiniella umbellata) was also studied. Mycelial growth rate, spore production and spore germination were significantly affected by media, temperature and isolates. In view of the present results, C. coronatus appears to be a good candidate for pest control in agricultural soils, as it has a wide tolerance to agricultural practices, has frequently been isolated from both vegetable and orchard fields, and is characterized by high mycelial growth rate, conidial production and conidial germination.     

河北省苹果园根际土壤中疑似致病镰孢菌种类

为了解引起河北省苹果再植病害的病原菌,在河北省10个地区苹果园中采集土壤样品,在实验室进行病原菌的诱集分离培养,根据形态和分子特征对主要病原菌进行种类鉴定。结果表明,在分离得到的293株真菌中,有116株镰孢菌,为分离频率最高的真菌。在形态学鉴定的基础上,对供试镰孢菌进行了分子鉴定。在基于核糖体基因内转录间隔区（rDNA-ITS）序列与翻译延长因子1α（EF-1α）序列片段构建的系统发育树中,代表菌株分别与GenBank登记的所属菌株聚于同一群。研究结果明确了河北省苹果再植病害的疑似致病镰孢菌,包括：尖孢镰孢Fusarium oxysporum、木贼镰孢F. equiseti、锐顶镰孢F. acuminatum、层出镰孢F. proliferatum和茄腐镰孢F. solani。     

Nocardia asteroides in the Soil of Kuwait

A pilot study was undertaken to determine the occurrence and distribution of pathogenic nocardiae in Kuwaiti soil. A total of 102 soil samples collected from two localities were investigated by the paraffin bait technique. Nocardia asteroides was the only species isolated from 42 (41%) soil samples. None of the isolates fulfilled the criteria required for identification of N. farcinica or N. nova. Thirty one (73.8%) isolates showed equivalent growth at 45 °C and 35 °C, 17 (40.4%) isolates utilized acetamide for carbon and nitrogen requirements and 3 (7.1%) isolates showed delayed arylsulphatase activity. Only a solitary isolate was resistant to cefamandole. Soil samples originating from the Kuwait University Campus Shuwaikh, which were rich in humus/organic matter, were more productive for N. asteroides (67%) than the samples which were devoid of it but were mixed with crude oil (39%). Sand samples that lacked organic matter and crude oil samples were least productive of N. asteroides. These preliminary findings do not suggest that massive oil contamination of soil in the Ahmadi oil field area during the Gulf war promoted the natural occurrence of N. asteroides. However, isolation of N. asteroides in as many as 41% of the soil sample is a significant observation warranting further epidemiologic studies including its possible role in the operation desert storm sickness syndrome. This is the first report on the natural occurrence of N. asteroides in Kuwait. This revised version was published online in June 2006 with corrections to the Cover Date.     

Detection of multiple mycetoma pathogens using fungal metabarcoding analysis of soil DNA in an endemic area of Sudan

Mycetoma is a tropical disease caused by several fungi and bacteria present in the soil. Fungal mycetoma and eumycetoma are especially challenging to treat; therefore, prevention, early diagnosis, and early treatment are important, but it is also necessary to understand the geographic distribution of these pathogenic fungi. In this study, we used DNA metabarcoding methodology to identify fungal species from soil samples. Soil sampling was implemented at seven villages in an endemic area of Sennar State in Sudan in 2019, and ten sampling sites were selected in each village according to land-use conditions. In total, 70 soil samples were collected from ground surfaces, and DNA in the soil was extracted with a combined method of alkaline DNA extraction and a commercial soil DNA extraction kit. The region for universal primers was selected to be the ribosomal internal transcribed spacer one region for metabarcoding. After the second PCR for DNA library preparation, the amplicon-based DNA analysis was performed using next-generation sequencing with two sets of universal primers. A total of twelve mycetoma-causative fungal species were identified, including the prime agent, Madurella mycetomatis, and additional pathogens, Falciformispora senegalensis and Falciformispora tompkinsii, in 53 soil samples. This study demonstrated that soil DNA metabarcoding can elucidate the presence of multiple mycetoma-causative fungi, which may contribute to accurate diagnosis for patient treatment and geographical mapping.     

Dematiaceous fungal pathogens isolated from nature

This study was conducted to demonstrate the presence of pathogenic dematiaceous fungi in nature. Using hamster and mouse inoculation techniques, 43 isolates of dematiaceous fungi were recovered from 39 samples of woody plant material and soil from the Virginia environment. Thirteen species were identified and included 4 Phialophora spp., 3 Cladosporium spp., 2 Exophiala spp., Sporothrix sp., Wangiella dermatitidis, Bispora betulina, and Scytalidium lignicola. Evidence is presented for the first isolations of C. trichoides from nature in the United States; these isolates proved to be pathogenic for mice in which they produced disease and death in a course similar to that seen in man. Natural isolates of Phialophora verrucosa, Phialophora repens, Exophiala jeanselmei, and Wangiella dermatitidis were identical to those species isolated from man using the following criteria: morphology, 12% gelatin reaction, and survival in laboratory animals.     

Factors affecting the occurrence and distribution of entomopathogenic fungi in natural and cultivated soils

Factors affecting the occurrence and distribution of entomopathogenic fungi in 244 soil samples collected from natural and cultivated areas in Spain were studied using an integrated approach based on univariate and multivariate analyses. Entomopathogenic fungi were isolated from 175 of the 244 (71.7 %) soil samples, with only two species found, Beauveria bassiana and Metarhizium anisopliae. Of the 244 soil samples, 104 yielded B. bassiana (42.6 %), 18 yielded M. anisopliae (7.3 %), and 53 soil samples (21.7 %) harboured both fungi. Log-linear models indicated no significant effect of habitat on the occurrence of B. bassiana, but a strong association between M. anisopliae and soils from cultivated habitats, particularly field crops. Also, irrespective of habitat type, B. bassiana predominated over M. anisopliae in soils with a higher clay content, higher pH, and lower organic matter content. Logistic regression analyses showed that pH and clay content were predictive variables for the occurrence of B. bassiana, whereas organic matter content was the predictive variable for M. anisopliae. Also, latitude and longitude predicted the occurrence of these same species, but in opposite directions. Altitude was found to be predictive for the occurrence of B. bassiana. Using principal component analysis, four factors (1 to 4) accounted for 86 % of the total variance; 32.8, 22.9, 19.6 and 10.4 % of the cumulative variance explained, respectively. Factor 1 was associated with high positive weights for soil clay and silt content and high negative weights for soil sand content. Factor 2 was associated with high positive weights for soil organic matter content and high negative weights for soil pH. Factor 3 was associated with high positive weights for latitude and longitude of the sampled localities and factor 4, had high positive weights only for the altitude. Bi-plot displays representing soil samples were developed for different factor combinations and indicated that, irrespective of geographical location, absence of both fungal species was determined by alkaline sandy soils with low organic matter content, whereas heaviness of soil texture, acidity and increasing organic matter content led to progressively higher percentages of samples harbouring entomopathogenic fungi. These results could aid decision-making as to whether or not a particular cultivated or natural soil is suitable for using entomopathogenic fungi as a pest control measure and for selecting the fungal species best suited to a particular soil.     

Distribution,occurrence and characterization of entomopathogenic fungi in agricultural soil in the Palestinian area

The occurrence of entomopathogenic fungi was investigated in irrigated vegetable fields and citrus orchards soils, over a nine-month period (April-December 1999),using the Galleria bait method (GBM). Entomopathogenic fungi were found to occur in 33.6% of the soil samples studied, with positive samples yielding 70 fungal isolates, belonging to 20 species from 13 genera. Conidiobolus coronatus was the most frequent and abundant entomopathogenic species recovered, comprising 31.4% of the total number of isolates. Soil pH, soil moisture content and the geographical location had minor or no effect on the isolation of entomopathogenic fungi in the fields studied. On the other hand, organic matter content of soil, and vegetation type were found to significantly affect the occurrence of entomopathogenic fungi in soil habitats, with orchard fields yielding larger numbers of isolates than vegetable fields. Using Koch's postulates the pathogenicity of fungal isolates to Galleria larvae was found to range from 16–100% (mortality rate). Isolates of C. coronatus proved to be the most virulent isolates recovered. The effect of media and temperature on mycelial growth rate, conidial production and conidial germination of six entomopathogenic fungal species (C. coronatus, Entomophaga grylli, Erynia castrans, Hirsutella jonesii, Paecilomyces farinosus and Sporodiniella umbellata) was also studied. Mycelial growth rate, spore production and spore germination were significantly affected by media, temperature and isolates. In view of the present results, C. coronatus appears to be a good candidate for pest control in agricultural soils, as it has a wide tolerance to agricultural practices, has frequently been isolated from both vegetable and orchard fields, and is characterized by high mycelial growth rate, conidial production and conidial germination.This revised version was published online in October 2005 with corrections to the Cover Date.     

Comparative study reveals unique features of the mycobiota in peat soils samples from Japan and Scotland

We isolated filamentous fungi from soil samples of peat layers in Aomori and Oita Prefectures in Japan and Perth and Kinross district in Scotland by a serial dilution plate technique. The mycobiota in each peat soil showed some common and characteristic features. The abundance of fungal isolates (CFU/g) from peat soil was low: about 1/3 to 1/30 compared with evergreen or coniferous forests or cultivated soil. Trichoderma or Mucorales species were scarcely observed; these fungi occupied only 3% of the total number of colonies. On the other hand, fungi such as Conioscypha and Tolypocladium that are normally isolated rather rarely were encountered at a comparatively high rate. Acremonium guillematii and Tolypocladium cylindrosporum were recorded for the first time in Japan. Sterile fungi occupied 50% of the total number of isolates. The low abundance of fast-growing fungi enabled us to pick slow-growing fungi up easily from the isolation medium. It is interesting that species not previously described in Japan, or scarcely reported, were isolated commonly from both Japanese and Scottish samples. A peat soil sample is therefore an attractive source of untapped microbial resources.     

Identification, phylogenetic analysis and characterization of obligate halophilic fungi isolated from a man-made solar saltern in Phetchaburi province, Thailand

A total of 17 species from 43 isolates were obtained through serial dilutions of soil samples isolated from one of the man-made solar salterns located in Ban Laem district of Phetchaburi province, Thailand. Soil analysis of the sample revealed high salinity and moisture content, slight alkalinity and low amounts of nitrogen, total organic carbon and organic matter in the habitat. Morphological analysis was performed on all isolates, and molecular identification and phylogenetic analysis were carried out only on the halophilic fungi isolated. Six halophilic fungi, belonging to four species, were identified among the isolates, including five strains of Aspergillus genus [Aspergillus flavus, A. gracilis, A. penicillioides (2 strains) and A. restrictus]. One species was found to be a yeast, namely, Sterigmatomyces halophilus, which was the most frequent isolate found among the halophilic fungi. All other isolates were halotolerant fungi. Characterization of the halophilic fungal isolates showed that they were best adapted to conditions of 10–15 % NaCl (w/v), slight alkalinity (pH 7.0–7.5) and a temperature range of 30–35 °C.     

湛江红树林滩涂可培养真菌种群多样性分析

【目的】明确湛江地区红树林滩涂海洋真菌的种类及其分布,为海洋真菌的开发利用研究奠定基础。【方法】运用稀释平板法从湛江市高桥及特呈岛红树林滩涂不同的潮位带(低、中、高)和不同树种(白骨壤、桐花树、木榄、红海榄)采集淤泥样品550份,采用真菌形态学和ITS序列分析技术进行多样性研究【。结果】分离获得海洋真菌274株,共鉴定出19属39种真菌,以曲霉属Aspergillus、青霉属Penicillium和木霉属Trichoderma真菌分离频率高,为湛江红树林滩涂优势真菌种类,尤其在中潮位带真菌种类最多。此外,分离获得真菌Talaromyces helicus,为中国新记录种。【结论】湛江红树林滩涂海洋真菌的种类十分丰富,具有潜在的开发和利用前景。     

Fluctuations in haemocyte density and microbial load may be used as indicators of fungal pathogenicity in larvae of Galleria mellonella

A positive correlation exists between the pathogenicity of bacteria and fungi when evaluated in the insect Galleria mellonella and mice. This work sought to determine whether fluctuations in the number of haemocytes and the proliferation of yeast cells in infected larvae could be used to determine the relative pathogenicity of a range of yeast isolates. Larvae were inoculated with 1 x 10(6) stationary-phase yeast cells and incubated in the dark at 30 degrees C for 48 h. The results indicated that larvae inoculated with the most pathogenic isolates (i.e. those capable of killing >80% of infected larvae) showed a significant reduction in haemocyte density. Larvae inoculated with isolates of low pathogenicity (i.e. capable of killing <20% of infected larvae) demonstrated only a small fluctuation in haemocyte numbers. The most pathogenic yeast isolates proliferated in the larvae, whereas the isolates of low pathogenicity did not. These results demonstrate a relationship between the ability of yeast isolates to kill larvae and changes in haemocyte density and yeast cell density in infected larvae. These end points may extend the applicability of the G. mellonella system for use with a wider range of microbial isolates.     

A ten years survey of Romanian soil screening for keratinophilic fungi (1958–1967)

Summary Out of 750 soil samples collected from various regions of Romania, during 1958–1967, a positive percentage of 86 was positive for keratinophilic fungi. Among the 13 isolated species, the most prevalent wereM. gypseum (54.6 %),C. keratinophilum (40.1 %) andK. (T.)ajelloi (27 %). BesideT. terrestre, found in 8.9 % of the examined soil samples, the percentage of the remaining species is low, not exceeding 4 %. In order of frequency:C. (T).evolceanui (3.2 %),M. cookei (2.2 %),T. mentagrophytes (2 %),T. quinckeanum (0.9 %),C. (T.)indicum (0.6 %). There have been also isolated, but only once;M. canis, A. multifidum, T. georgiae andT. vanbreuseghemii.The studies carried out on the immunobiological properties of the Romanian soil isolates (including intracutaneous tests, serological reactions and therapeutic efficiency), have showed a close relationship between these fungi and the common pathogenic dermatophytes. The pathogenic power of the keratinophilic fungi is briefly discussed.