A First Record of Obligate Halophilic Aspergilli from the Dead Sea

The isolation of obligate halophilic aspergilli from the Dead Sea and the range of salt tolerance of halophilic fungi isolated, are reported here for the first time. The mycobiota of the Dead Sea isolated in this study, was dominated by Aspergillus and Penicillium species; Cladosporium were found in lesser numbers. All three genera were obtained from the water sample; however, Aspergillus was the only genus obtained from the sediment. There was significant difference in growth of each isolate at different salt concentrations and intraspecies analysis revealed dissimilarity in response of strains to different salt concentrations in the growth medium The isolates were euryhaline, with halotolerance up to 20–25% solar salt, Aspergillus and Penicillium species showing a higher level of halotolerance, as compared to that of Cladosporium. Halophilic fungi were found in greater numbers in the sediment sample as compared to that in the water sample. Penicillium and Cladosporium species were exclusively facultative halophiles, while some species of Aspergillus were facultative halophiles. All the obligate halophiles isolated, belonged to the genus Aspergillus and were identified as A. penicillioides and A unguis, the latter being a first record of the species from the Dead Sea.     

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.     

Biosurfactant Production by Antarctic Facultative Anaerobe <Emphasis Type="Italic">Pantoea</Emphasis> sp. During Growth on Hydrocarbons

The facultative anaerobe Pantoea sp. strain A-13, isolated from ornithogenic soil of Dewart Island (Frazier Islands), Antarctica, produced glycolipid biosurfactants when grown on n-paraffins or kerosene as the sole source of carbon and energy. Hemolysis of erythrocytes, growth inhibition of Bacillus subtilis, and thin-layer chromatography studies have suggested that the secreted glycolipids are rhamnolipids. Glycolipids produced by kerosene-grown cells decreased the surface tension at the air–water interface to 30 mN/m and possessed a low critical micelle concentration value of 40 mg/l, which indicated high surface activity. They efficiently emulsified aromatic hydrocarbons, kerosene, and n-paraffins. Biosurfactant production contributed to an increase in cell hydrophobicity, which correlated with increased growth of the strain on tested hydrocarbons. According to the results, the Antarctic biosurfactant-producing strain Pantoea sp. A-13 appears to be valuable source for application in accelerated environmental bioremediation.     

Fungal microbiota dynamics as a postmortem investigation tool: focus on Aspergillus,Penicillium and Candida species

Aims: To investigate the presence of fungi during three human decomposition stages: bloated, putrefaction and skeletonization. Methods and Results: The samples were gathered in the city of Fortaleza, Ceará, Brazil, from the public morgue and cemeteries. The material was submitted to conventional mycological procedures by direct examination and macro/micro morphological and biochemical analyses. The main fungi isolated were Aspergillus spp., Penicillium spp. and Candida spp. in the bloated stage (n = 34 cadavers) and in the putrefaction stage (n = 6 cadavers), while in the skeletonization stage (n = 20 cadavers), the main fungi were Aspergillus spp., Penicillium spp. and Mucor sp. Conclusions: Aspergillus, Penicillium and Candida species were associated with decomposed human cadavers. Significance and Impact of the Study: These findings enable tracing out the profile of fungal communities of human cadavers for the first time. However, much more research will be necessary to develop this new segment of mycology and to enable its routine use in forensic science.     

Design of bacterial defined mixed cultures for biodegradation of specific crude oil fractions, using population dynamics analysis by DGGE

Hydrocarbon-degrading bacteria isolated from oil-polluted soils, were used to design three defined mixed cultures (DMC) for biodegradation of Maya crude oil fractions. The first degrading culture, DMC A was made up with 10 strains. Design of DMC B (six strains) and DMC C (three strains) was based on DGGE profiles obtained throughout biodegradation assays of different petroleum fractions. Biodegradation of the aliphatic fraction (10 000 mg l⁻¹) and an aromatic–polar mixture (5000 mg l⁻¹) was evaluated for the DMC B. Biodegradation of total hydrocarbons (10 000 mg l⁻¹) and its fractions was evaluated for DMC B and DMC C. During biodegradation assays, O₂ consumption and CO₂ production were assessed by respirometry, while population dynamics of predominant strains was based on PCR-DGGE profiles of partial 16S rDNA. Aliphatic fraction was completely biodegraded by DMC B, while degradation of the aromatic–polar mixture was 12.5% and for total hydrocarbons 40.5%. DMC B was able to degrade the aromatic fraction (31%) and even the polar fraction (19.6%) present in total hydrocarbons. DMC C degraded the aromatic and polar fractions (5.6% and 2%, respectively) present in total hydrocarbons. DGGE profiles of the DMCs indicated that Pseudomonas sp., Gordonia rubripertincta and a non-identified strain were predominant and probably responsible of the hydrocarbons biodegradation. The use of DGGE-fingerprinting to track microbial populations, allowed selecting strains to design efficient oil-degrading defined mixed cultures.     

The air spora close to a compost facility in Northeast Oklahoma: Part I—spore trap sampling

A compost facility in northeast Oklahoma is located relatively close to a residential area and is the focus of complaints about smell and concerns about health effects. Several species of Aspergillus have been known to cause health problems, and at least one of these species is dominant in compost. The atmosphere surrounding the compost facility was monitored for 1 year using Burkard spore traps to determine if there was a significant difference in Penicillium/Aspergillus type spores concentration between a test and control site. Samplers were situated 710 m downwind for the test site and 6,085 m upwind at the control site. There was no significant difference in mean concentration of Penicillium/Aspergillus type spores between the two sites (t = 0.576 P > 0.05). The mean concentration of total spores was significantly higher at the upwind control site (t = −7.64, P < 0.01). Wind direction was examined to determine if the compost facility was a possible source for any spikes in concentration. No clear relationship was found between wind direction and mean Penicillium/Aspergillus concentration at the test site, but peak concentrations of Penicillium/Aspergillus seen at the test site were on days when it was downwind from the composting facility. However, these concentrations were no higher than those seen at the control site on other days. If the compost was releasing large amounts of Penicillium/Aspergillus type spores into the atmosphere they were generally diluted to background levels by the time they reached the test site.     

Biological transformation of thiourea

Summary Six strains of fungi: two Aspergilli of the A. glaucus group, Penicillium sp. Trichoderma viride, and two unidentified dark fungi, were found able to use thiourea as a source of nitrogen. One of the Aspergilli and the Penicillium showed the best utilization, but the growth was always slow, and no more than 50% (usually much less) of the thiourea-nitrogen was recovered in the mycelium.Assimilation of thiourea-nitrogen by Aspergillus and Penicillium was accompanied by oxidation of the sulphur to sulphate in roughly approximate stoichiometric proportion. The same fungi were able to produce sulphate from cystine as combined source of nitrogen and sulphur.     

Studies on the Utilization of Hydrocarbons by Microorganisms

Many strains of the hydrocabon-utilizing yeasts were isolated from various kinds of natural sources by accumulation culture.

Among those yeasts, two strains, S315YI and S131YI, which were identified with Candida tropilcais, assimilated hydrocarbons abundantly. As for type cultures, it was found that many strains of them could utilize hydrocarbons too, especially the strains which belonged to Genus Candida. However, as regards to the ability in utilizing hydrocarbons, no yeast from type culture collections utilized hydrocarbons better than the yeasts newly isolated from nature. Addition of the natural nutrients such as corn steep liquor to the cultural broth of Strain S315Y1 showed no effect on the production of yeast cells.

The yeast Strain S315Y1 assimilated the higher boiling points fraction of n-paraffins in comparison with the hydrocarbon-utilizing bacterium, Pseudomonas aeruginosa S7B1, which had been reported by the authors. Ribonucleic acid contents of dried cells of the yeast Strain S315Y1 and S131Y1 were 5.3 and 4.4% respectively by Schmidt-Thannhauser-Schneider method.     

Immunological relationship of extra-cellular polysaccharide antigens produced by different mould species

Moulds are able to produce extracellular polysaccharide antigens which are heat-stable and almost genus specific. Of 44 different strains of Penicillium 41 (93%) and all 12 strains of Aspergillus tested produced detectable quantities of an immunologically related antigen. Additionally 10 of these 56 strains produced an antigen immunologically related to the antigen produced by the genera Mucor and Fusarium. Immunologically different, but genus-specific antigens were produced by each of the species belonging to the genus Geotrichum, Fusarium, Cladosporium, Mucor and Rhizopus. The antigens produced by Mucor and Rhizopus, however, were immunologically related.     

Secondary metabolites in taxonomy of the <Emphasis Type="Italic">Penicillium</Emphasis> fungi

A correlation was established between species specificity and synthesis of specific secondary metabolites by the Penicillium fungi. Strains of the subgenus Aspergilloides usually synthesize metabolites of polyketide nature. Most strains of the subgenus Furcatum produce clavine ergot alkaloids and metabolites of diketopiperazine nature. The only clavine ergot alkaloids and diketopiperazine alkaloids produced by strains of the subgenus Biverticillium are rugulovasines and rugulosuvines, respectively. Species designations of the strains of the subgenus Penicillium isolated from permafrost soil, the Mir orbital complex, and sites undergoing anthropogenic load were refined based on the marker secondary metabolites. Changes in the taxonomic position of some strains in the genus Penicillium are suggested.     

Aspergillus on tree nuts: incidence and associations

California exports tree nuts to countries where they face stringent standards for aflatoxin contamination. Trade concerns have stimulated efforts to eliminate aflatoxins and Aspergillus flavus from almonds, pistachios and walnuts. Incidence of fungi on tree nuts and associations among fungi on tree nuts were studied. Eleven hundred pistachios, almonds, walnuts and brazil nuts without visible insect damage were plated on salt agar and observed for growth of fungi. Samples came both from California nut orchards and from supermarkets. To distinguish internal fungal colonization of nuts from superficial colonization, half the nuts were surface-sterilized before plating. The most common genera found were Aspergillus , Rhizopus and Penicillium . Each species of nut had a distinct mycoflora. Populations of most fungi were reduced by surface sterilization in all except brazil nuts, suggesting that they were present as superficial inoculum on (rather than in) the nuts. In general, strongly positive associations were observed among species of Aspergillus ; nuts infected by one species were likely to be colonized by other species as well. Presence of Penicillium was negatively associated with A. niger and Rhizopus in some cases. Results suggest that harvest or postharvest handling has a major influence on nut mycoflora, and that nuts with fungi are usually colonized by several fungi rather than by single species. This revised version was published online in June 2006 with corrections to the Cover Date.     

Induction and Citric Acid Productivity of Fluoroacetate-sensitive Mutant Strains of Candida lipolytica

To establish a novel process for the economical production of citric acid from n-paraffins by yeast, attempts were made to obtain some mutant strains capable of producing citric acid in higher yield without (+)-isocitric acid.

From among the mutant strains derived from Candida lipolytica ATCC 20114, which produced citric acid and (+)-isocitric acid in the ratio of about 60:40 from n-paraffins, a citrate non-utilizing mutant strain, K-20, and a fluoroacetate-sensitive mutant strain , S-22, were selected on the basis of high citric acid and low (+)-isocitric acid productivity.

The mutant strain S-22 showed extremely poor growth in a medium containing sodium citrate as the sole carbon source and extremely high sensitivity to fluoroacetate. The production ratio of citric acid and (+)-isocitric acid by the mutant strain was changed to 97:3, and the yield of the citric acid from n-paraffins, charged to the fermentation medium, reached 145%(w/w).     

Production of Proteinase on Noncarbohydrate Carbon Sources by Pseudomonas aeruginosa

Proteinase production by Pseudomonas aeruginosa was studied in medium containing noncarbohydrate materials, especially various hydrocarbons, as the sole carbon source. On heavy oil, kerosene, n-paraffinic hydrocarbon of C₁₂, C₁₄, or C₁₆, and propylene glycol, the bacteria grew well and high protinase production was observed. However, production on paraffinic hydrocarbon differed remarkably with strains of varied origins. The elastase-positive strain, IFO 3455, showed abundant growth and high proteinase production on medium containing a paraffin of C₁₂, C₁₄, or C₁₆, whereas the elastase-negative strain, IFO 3080, showed little growth on the same medium. Neither elastase-positive nor elastase-negative strains, however, utilized n-paraffins of C₅ to C₁₀, or various aromatic hydrocarbons such as benzene, naphthalene, phenanthrene, and anthracene. The proteinases produced on the noncarbohydrate medium were identical with those produced in glucose medium.     

Characterization of fungi isolated from the equipment used in the International Space Station or Space Shuttle

As a part of a series of studies regarding the microbial biota in manned space environments, fungi were isolated from six pieces of equipment recovered from the Japanese Experimental Module “KIBO” of the International Space Station and from a space shuttle. Thirty‐seven strains of fungi were isolated, identified and investigated with regard to morphological phenotypes and antifungal susceptibilities. The variety of fungi isolated in this study was similar to that of several previous reports. The dominant species belonged to the genera Penicillium, Aspergillus and Cladosporium, which are potential causative agents of allergy and opportunistic infections. The morphological phenotypes and antifungal susceptibilities of the strains isolated from space environments were not significantly different from those of reference strains on Earth.     

Aflatoxin and aflatoxicosis

A wide range of fungi, amounting to fifty-six species belonging to twenty-two genera, have been recovered from animal feeds and foodstuffs.The most frequent fungi were Aspergillus niger, and A. flavus, followed by Mucor racemosus, Alternaria alternata, Rhizopus stolonifer, Penicillium corylophilum and P. notatum. Three genera were found to be of moderate occurrence, namely, Mucor, Rhizopus and Alternaria. The three following genera were of low occurrence: Cladosporium, Fusarium, and Neurospora.The fluorescence method of detecting aflatoxin-producing strains demonstrated that only one isolate of A. flavus possesses this property. Certain species of Penicillium and Aspergillus produced fluorescent substances (metabolites) similar in color to B and G aflatoxin. These substances were subsequently proved not to be aflatoxin by (TLC) chromatography.The animal and public health significance from such toxins was also discussed.     

Aspergillus and other moulds in the air of Kuwait

A one-year survey was carried out to study the aerial prevalence of Aspergillus species and other moulds in the outdoor and indoor environments of Kuwait. Petri plates containing rose-Bengal medium were exposed for 20 minutes twice a month using a six-stage Andersen air sampler at the pre-determined sites. The exposed plates were incubated at 28 °C ± 1 °C up to 5 days and colonies were enumerated and identified by colonial and microscopic morphology. The data revealed that Aspergillus species were the predominant component (27.7%) of the outdoor aerospora of Kuwait and A. fumigatus alone accounted for 21.3% of the total aspergilli. In contrast, Cladosporium species formed the major component of the indoor aerospora (22.8%), followed by Aspergillus species (20.9%), Penicillium species (14.6%), and Bipolaris species (10.6%). A comparison of the fungi recorded in the outdoor and in the indoor air revealed that Aspergillus, Alternaria and Fusarium were significantly higher in the outdoor environment, whereas Cladosporium, Penicillium, and Bipolaris were significantly higher in the indoor environment. The relative prevalence of Aspergillus species and other moulds in the outdoor and indoor air of Kuwait was as follows: A. fumigatus 5.9 and 9.8%, A. flavus 4.9 and 3.9%, other aspergilli 16.8 and 7.0%, Alternaria species 19.8 and 7.9%, Cladosporium species 13.7 and 22.8%, Penicillium species 7.6 and 14.6%, and other moulds 31.2 and 34.1%, respectively. During the study, 25 different genera were identified, indicating a wide diversity in the spectrum of local fungal aerospora. The study provides useful information on the prevalence of allergenic fungi in the outdoor and indoor environments of Kuwait. This revised version was published online in June 2006 with corrections to the Cover Date.     

Low occurrence of patulin‐ and citrinin‐producing species isolated from grapes

Aims: To assess the ability of fungi isolated from grapes to produce patulin and citrinin. Methods and Results: A total of 446 Aspergillus isolates belonging to 20 species and 101 Penicillium isolates were inoculated in Czapek yeast extract agar and yeast extract sucrose agar and incubated for 7 days at 25°C. Extracts were analysed for patulin and citrinin by thin‐layer chromatography. None of the isolates of Aspergillus spp. produced either patulin or citrinin. Patulin was produced by three isolates of Penicillium expansum and two of Penicillium griseofulvum. Citrinin was produced by five isolates of P. expansum, two of Penicillium citrinum and one of Penicillium verrucosum. Conclusions: Our results show that the Aspergillus and Penicillium species commonly isolated from grapes are not a source of the mycotoxins, patulin and citrinin. Significance and Impact of the Study: The possibility of co‐occurrence of patulin and citrinin with ochratoxin A in grapes and grape products remain low, owing to the low frequency of isolation of potentially producing species.     

The potential for hydrocarbon biodegradation and production of extracellular polymeric substances by aerobic bacteria isolated from a Brazilian petroleum reservoir

Extracellular polymeric substances (EPS) can contribute to the cellular degradation of hydrocarbons and have a huge potential for application in biotechnological processes, such as bioremediation and microbial enhanced oil recovery (MEOR). Four bacterial strains from a Brazilian petroleum reservoir were investigated for EPS production, emulsification ability and biodegradation activity when hydrocarbons were supplied as substrates for microbial growth. Two strains of Bacillus species had the highest EPS production when phenanthrene and n-octadecane were offered as carbon sources, either individually or in a mixture. While Pseudomonas sp. and Dietzia sp., the other two evaluated strains, had the highest hydrocarbon biodegradation indices, EPS production was not detected. Low EPS production may not necessarily be indicative of an absence of emulsifier activity, as indicated by the results of a surface tension reduction assay and emulsification indices for the strain of Dietzia sp. The combined results gathered in this work suggest that a microbial consortium consisting of bacteria with interdependent metabolisms could thrive in petroleum reservoirs, thus overcoming the limitations imposed on each individual species by the harsh conditions found in such environments.     

Microbial degradation of petroleum hydrocarbons in a polluted tropical stream

Summary Crude oil degradation was observed in water samples from three sites along the course of a polluted stream in Lagos, Nigeria. Consistent increase and decrease in the total viable counts (TVCs) of indigenous organisms occurred in the test and control experiments, respectively. Enrichments of the water samples with crude oil resulted in the isolation of nine bacteria belonging to seven genera. A mixed culture was developed from the assemblage of the nine species. The defined microbial consortium utilized a wide range of pure HCs including cycloalkane and aromatic HCs. Utilization of crude oil and petroleum cuts, i.e., kerosene and diesel resulted in an increase in TVC (till day 10) concomitant with decreases in pH and residual oil concentration. Crude oil, diesel and kerosene were degraded by 88, 85 and 78%, respectively, in 14 days. Substrate uptake studies with axenic cultures showed that growth was not sustainable on either cyclohexane or aromatics while degradation of the petroleum fractions fell below 67% in spite of extended incubation period (20 day). From the GC analysis of recovered oil, while reductions in peaks of n-alkane fractions and in biomarkers namely n-C₁₇/pristane and n-C₁₈/phytane ratios were observed in culture fluids of pure strains, complete removal of all the HC components of kerosene, diesel and crude oil including the isoprenoids was obtained with the consortium within 14 days.