Allergenic and mutagenic characterization of 14 <Emphasis Type="Italic">Penicillium</Emphasis> species

Extracts of 14 Penicillium species, P. aurantiogriseum, P. brevicompactum, P. citrinum, P. chrysogenum, P. expansum, P. glabrum, P. hirsutum, P. italicum, P. janthinellum, P. melini, P. oxalicum, P. purpurescens, P. simplicissimum, and P. viridicatum were investigated by total protein, specific enzyme determinations, isoelectric focusing (IEF), sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using pooled human atopic IgE. Considerable variation was observed between the Penicillium species with respect to protein yield and the number of distinct protein bands resolved in IEF. Using the Api-Zym system, the most common activities observed among the extracts included acid and alkaline phosphatase, phosphodiamidase and β-glucosaminidase. The number of discrete atopic IgE-reactive bands in immunoblots of Penicillium extracts ranged from 1 (P. chrysogenum) to 9 (P. viridicatum). Certain allergens showed potential for cross-reactivity between species, including 52 and 54 kDa proteins in P. citrinum, P. purpurescens, P. viridicatum and 40 kDa proteins in several species. The extracts were also nonmutagenic when tested with the Ames assay using Salmonella strains TA98 and TA100. The results tend to indicate that P. viridicatum, P. janthinellum, P. oxalicum, P. brevicompactum and P. italicum, which are highly immunogenic as well as allergenic, could possibly be good candidates for allergen cloning studies through the construction of cDNA libraries. The extracts were non-mutagenic and can be used safely for skin testing.*Presented in part at the 7th International meeting of Aerobiology, Montebello, August, 2002.     

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.     

Penicillium Mycobiota in Arctic Subglacial Ice

Fungi have been only rarely isolated from glacial ice in extremely cold polar regions and were in these cases considered as random, long-term preserved Aeolian deposits. Fungal presence has so far not been investigated in polar subglacial ice, a recently discovered extreme habitat reported to be inhabited exclusively by heterotrophic bacteria. In this study we report on the very high occurrence (up to 9000 CFU L⁻¹) and diversity of filamentous Penicillium spp. in the sediment-rich subglacial ice of three different polythermal Arctic glaciers (Svalbard, Norway). The dominant species was P. crustosum, representing on the average half of all isolated strains from all three glaciers. The other most frequently isolated species were P. bialowiezense, P. chrysogenum, P. thomii, P. solitum, P. palitans, P. echinulatum, P. polonicum, P. commune, P. discolor, P. expansum, and new Penicillium species (sp. 1). Twelve more Penicillium species were occasionally isolated. The fungi isolated produced consistent profiles of secondary metabolites, not different from the same Penicillium species from other habitats. This is the first report on the presence of large populations of Penicillium spp. in subglacial sediment-rich ice.     

Dominant fungi in the rhizosphere of established tea bushes and their interaction with the dominant bacteria under in situ conditions

Species of Penicillium and Trichoderma were found to dominate the rhizosphere of established tea bushes in a detailed study conducted from various tea growing locations in India. Penicillium erythromellis, P. janthinellum, P. raistrickii, Trichoderma pseudokoningii and T. koningii were found to be closely associated with tea roots. While seasonal fluctuation was observed in the case of Penicillium spp., the population of Trichoderma spp. showed less variation during the year. Both species were sensitive to low temperatures. In general, fungi associated with the tea rhizosphere were found to prefer a mesophillic temperature range (15 °C to 35 °C). The dominant species of Penicillium and Trichoderma also exhibited tolerance to lower temperatures, i.e., 5 to 10 °C on agar plates. Most fungi were able to grow in a wide range of pH (4 to 12). Lowering of soil pH in the rhizosphere of tea bushes was positively correlated with the age of the bush and may have affected the development of a specific microbial community in the rhizosphere.The populations of Penicillium and Trichoderma species were inversely correlated with the populations of two most dominant rhizosphere bacteria, Bacillus subtilis and B. mycoides. Both Bacillus species have been shown to have antagonistic activity against these two fungi under in vitro conditions. The present study demonstrates the existence of a similar antagonism under in situ conditions in the rhizosphere of established tea bushes.     

青霉属的三个新种

本文报导青霉属的三个新种,它们不同于此属已发表的诸种.异型青霉(Penicillium heteromorphum Kong et Qi),根据帚状枝的结构和分生孢子的某些特征,近于两型孢青霉(Penicillium dimorphosporum Swart),但此种在查氏琼脂上生长很稀疏,并有两种大小的分生孢子,大分生孢子直径可达10 pm,壁光滑,在观察菌落期间,有时大分生孢子落在基质上,很快萌发生长成简单的分生孢子结构,这些特征易于与后者区分.结节青霉(Penicillium nodulum Kong et Qi)在某些方面类似纠缠青霉(Penicilliumimplicatum Biourge),此新种的菌落特征,柄顶端膨大似顶囊,呈结节状膨大的营养菌丝等特征而不同于纠缠青霉.神农架青霉(Penicillium shennong jianum Kong et Qi)和毡毛青霉(Penicillium velutinum van Heyma)相近,而新种在多种培养基上生长都局限,菌落面呈带褐的橄榄色,分生孢子壁光滑等特征可与毡毛青霉区分开来.     

NephrotoxigenicPenicillium species occurring on farm-stored cereal grains in western Canada

The incidence of nephrotoxigenicPenicillium species on farm-stored cereals in western Canada was determined by morphological and metabolite profile examination. Of the 142 isolates examined 102 were toxin producers with 61P. aurantiogriseum and 27P. freii. Other nephrotoxigenic species includedP. tricolor (6 isolates),P. verrucosum Chemotype II (4 isolates) andP. viridicatum Westling (4 isolates). The nephrotoxigenicPenicillium species profile for western Canada appears to differ from that of Denmark whereP. verrucosum,P. cyclopium,P. freii and, to a lesser extent,P. aurantiogriseum,P. polonicum, andP. viridicatum predominate.     

Potency of plant extracts against Penicillium species isolated from different seeds and fruits in Saudi Arabia

Antifungal activity of extracts of cinnamon (Cinnamomum zeylanicum), Cloves (Syzygium aromaticum), ginger (Zingiber officinale) and turmeric (Curcuma longa) were evaluated in vitro against 17 Penicillium spp. Seed disease and rotten fruit caused by these species cause considerable loss of quality for different agricultural products. Isolates of Penicillium spp. were screened for production of patulin an important serious mycotoxin. About 70.59% of Penicillium spp. produced this toxin in concentrations ranging from 4 to 31 ppb. The response of Penicillium spp.to plant extracts differed according to the plant extract and concentration. Cinnamon extract showed the greatest effect on P. asperosporum, P. aurintogriseum and P. brevicompactum, and cloves extract produced the greatest effect on P. chermesinum and P. duclauxii. Turmeric extract had less effect on P. duclauxii. Cloves extract was the most effective in reducing the growth of Penicillium spp. On the other hand, ginger extract with all concentrations used had less effect against most Penicillium spp in the laboratory. Plant extracts are promising as natural sources of environmentally friendly compounds in laboratory studies.     

Characterization of <Emphasis Type="Italic">Penicillium</Emphasis> Species by Ribosomal DNA Sequencing and BOX,ERIC and REP-PCR Analysis

The genus Penicillium is one of the largest and widely distributed fungal genera described to date. As a result, its taxonomic classification and species discrimination within this genus has become complicated. In this study, 52 isolates that belonged to the Penicillum genus and other related genera were characterized using two DNA-based methods: (i) analysis of the nucleotide sequences of internal transcribed spacers in ribosomal DNA and (ii) analysis of DNA fingerprints that were generated by polymerase chain reactions with specific primers for enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP) sequences, and BOX elements. Using both methods, Penicillium species were discriminated from other fungal genera. Furthermore, Penicillium species that include strains which are used as biocontrol agents, such as P. glabrum, P. purpurogenum, and P. oxalicum, could be distinguished from other Penicillium species using these techniques. Based on our findings, we propose that a polyphasic approach that includes analysis of the nucleotide sequences of ribosomal DNA and detecting the presence of highly conserved, repeated nucleotide sequences can be used to determine the genetic relationships between different Penicillium species. Furthermore, we propose that our results can be used as a start point to develop a strategy to monitor the environmental presence of particular strains of Penicillium species when they are used as biocontrol agents.     

Effects of Penicillium extracts on germination vigour in subsequent seedling growth of tomato (Solanum lycopersicum L.)

Penicillium spp. are well known to produce a variety of beneficial metabolites for plant growth and survival, as well as they defend their hosts from attack of certain pathogens. In this study, effects of culture filtrate of different Penicillium spp. were tested on tomato seeds. On the whole, presoaking of seeds in the filtrates of the nine Penicillium isolates tested significantly increased seed germination when compared with the control seeds. Cultural extracts of P. expansum and P. bilaii were highly effective in growth promotion up to 90%. It was also observed that P. implicatum and P. oxalicum significantly enhanced the root growth in tomato seedling as compared to other species. In case of shoot length, P. verrucosum (3.38 cm), P. granulatum (2.81 cm) and P. implicatum (2.62 cm) were effective. P. implicatum was quite promising in increasing shoot and root length in tomato seedlings, while P. simplicissimum and P. citrinum were less effective on seedling growth. The plant growth-promoting ability of Penicillium strains may help in growth promotion in other plants and crops. Penicillium spp. are already known for producing various mycotoxins and enzymes. Plant growth-promoting ability of Penicillium spp. will open new aspects of research and investigations. The role of Penicillium spp. in tomato plant growth requires further exploration.     

A report on Penicillium in the intramural and extramural air of residential areas of Nagpur city (India)

Prevalence of different species of Penicillium and their concentrations per cubic meter of air were evaluated with the use of Hi-Air sampler system Mark II (Hi-Media Laboratories Ltd., India) in the air of homes (bed-rooms) at four different sites in Nagpur. At each of these sites, air sampling was done fortnightly in triplicate for 2 years duration from June 2000 to May 2002. The sampling was also done in triplicate for the outdoor air in the vicinity of each home on the same day immediately after the indoor sampling was over. The mean concentration of Penicillium colony forming units at four different sites in the indoor air was 32, 46.9, 35 and 35.4 CFU/m³, respectively, whereas in the outdoor air at these same four sites, the mean concentration was 24, 28, 25 and 25.8 CFU/m³ respectively. The Penicillium concentration in the indoor air was found to be higher in winter than in other seasons (ANOVA, p < 0.05). Concentration of Penicillium spp. in intramural environment was always higher than that in extramural environment. Statistically significant difference existed between intramural and extramural environments at all the sites, with maximum difference at a site, which is old crowded area of the city. During the 2-years investigations, 11 species of Penicillium were isolated from the indoor air while nine species were isolated from the air outside the homes. The dominant species of Penicillium in indoor as well as outdoor air were P. citrinum (33.78 and 32.81), P. oxalicum (19.70 and 22.60), and P. chrysogenum (17.64 and 14.50). The percentage of the Penicillium in the indoor air was 10.70 while it was 8.36 in outdoor air. Indoor air showed the presence of P. glaber and P. sclerotiorum, which were absent in the outdoor air.     

Management of post-harvest fruit spoilage fungi by some potential spice extracts

The experiment was carried out to evaluate the antifungal potential of aqueous and ethanolic extracts of four spices (Allium sativum, Zingiber officinale, Cinnamomum zeylanicum and Capsicum annuum) against the important post-harvest spoilage fungi isolated from diseased fruits. In total, 276 isolates of post-harvest spoilage fungi were isolated from four different fruit types (Persea americana, Musa acuminata, Citrus sinensis and Lycopersicon esculentum), out of which 183 isolates were identified while 93 isolates were remain unidentified. The most dominant post-harvest spoilage fungus was Rhizopus sp. (26.45%), followed by Penicillium sp. (19.93%), Aspergillus sp. (10.86%) and Fusarium sp. (9.06%). Results of disc diffusion assay showed that ethanolic extract of C. zeylanicum was found to be most effective against Penicillium sp., followed by aqueous extracts of A. sativum. The ethanolic extract of C. zeylanicum in agar amended assay and minimal inhibitory concentrations were found to be very efficient (100% inhibition) against all the tested fungi. Results of in vivo study showed that pre-inoculated C. zeylanicum ethanolic extract and A. sativum aqueous extract were found effective in reducing the disease severity (6.24–13.67%) and (7.91–13.15%) against the Penicillium, Rhizopus, Aspergillus and Fusarium sp.     

Antagonism against Rhizoctonia solani and fungitoxic metabolite production by some Penicillium isolates

A number of Penicillium isolates were recovered in association to Rhizoctonia solani strains pathogenic on tobacco and from soil on plates pre-colonized by the pathogen itself. Their antagonism toward R. solaniAG-2-1 was evaluated in dual cultures in vitro. Inhibition of growth was evident to some extent in most pairings, while hyphal interactions referable to mycoparasitic relationships were not observed. However, the occurrence of plasmolysis and/or vacuolisation and the induction of monilioid cells were indicative of the release of bioactive compounds. Therefore, production of fungitoxic metabolites was tested by adding concentrated culture filtrates of each Penicillium isolate to the growth medium of R. solani. Complete and lasting inhibition was incited by culture filtrates of some isolates belonging to P. brevicompactum, P. expansum, and P. pinophilum. Three purified compounds, respectively mycophenolic acid, patulin and 3-O-methylfunicone, which were extracted from culture filtrates, were able to inhibit R. solani in vitro. Their production was also detected in dual cultures of the same Penicilliumstrains with R. solani prepared in sterilized soil and when the Penicilliumstrains were cultured directly on R. solani mycelium harvested from liquid cultures. The possible role of such metabolites in antagonism of the above-mentioned Penicilliumspecies against R. solani is discussed.     

Antifungal effect of Penicillium metabolites against some fungi

Microorganisms are increasingly exploited as a source of new biological control agents. Genus Penicillium is a source of novel bioactive molecules which can be used as antifungal agents. The objective of this study was to evaluate the antifungal potential of Penicillium strains. Culture filtrates of two Penicillium species were tested for their antifungal potential by well diffusion assays. Filtrate of Penicillium isolates showed high antifungal effects on mycelial growth of Fusarium oxysporum, Fusarium solani, Macrophomina phaseolina, Aspergillus japonicus var aculeatus and Cladosporium cladosporioides. But Penicillium italicum inhibit the fungal growth from 45 to 68% as compared to Penicillium simplissimum (25–68%). However in case of A. japonicus var aculeatus, Penicillium spp. extracts were equally effective and reduce the colony growth up to 68%. However, P. simplissimum extract was least effective in case of M. phaseolina, where it decreased the colony growth only 25%.     

The identity of <Emphasis Type="Italic">Penicillium</Emphasis> sp. 1, a major contaminant of the stone chambers in the Takamatsuzuka and Kitora Tumuli in Japan,is <Emphasis Type="Italic">Penicillium paneum</Emphasis>

Penicillium appeared as the major dweller in the Takamatsuzuka Tumulus (TT) and Kitora Tumulus (KT) stone chambers, both located in the village of Asuka, Nara Prefecture, in relation to the biodeterioration of the 1,300-year-old mural paintings, plaster walls and ceilings. Of 662 Penicillium isolates from 373 samples of the TT (sampling period, May 2004–2007) and the KT (sampling period, June 2004–Sep 2007), 181 were phenotypically assigned as Penicillium sp. 1 which shared similar phenotypic characteristics of sect. Roqueforti in Penicillium subg. Penicillium. Fifteen representative isolates of Penicillium sp. 1, 13 from TT and 2 from KT, were selected for molecular phylogenetic analysis. The 28S rDNA D1/D2, ITS, β-tubulin, and lys2 gene sequence-based phylogenies clearly demonstrated that the three known species P. roqueforti, P. carneum and P. paneum in sect. Roqueforti, and all TT and KT isolates grouped together. In addition to this, TT and KT isolates formed a monophyletic group with the ex-holotype strain CBS 101032 of P. paneum Frisvad with very strong bootstrap supports. So far, P. paneum has been isolated only from mouldy rye breads, other foods, and baled grass silage. Therefore, this is the first report of P. paneum isolation from samples relating to the biodeteriorated cultural properties such as mural paintings on plaster walls.     

A report on <Emphasis Type="Italic">Penicillium</Emphasis> in the intramural and extramural air of residential areas of Nagpur city (India)

Prevalence of different species of Penicillium and their concentrations per cubic meter of air were evaluated with the use of Hi-Air sampler system Mark II (Hi-Media Laboratories Ltd., India) in the air of homes (bed-rooms) at four different sites in Nagpur. At each of these sites, air sampling was done fortnightly in triplicate for 2 years duration from June 2000 to May 2002. The sampling was also done in triplicate for the outdoor air in the vicinity of each home on the same day immediately after the indoor sampling was over. The mean concentration of Penicillium colony forming units at four different sites in the indoor air was 32, 46.9, 35 and 35.4 CFU/m³, respectively, whereas in the outdoor air at these same four sites, the mean concentration was 24, 28, 25 and 25.8 CFU/m³ respectively. The Penicillium concentration in the indoor air was found to be higher in winter than in other seasons (ANOVA, p < 0.05). Concentration of Penicillium spp. in intramural environment was always higher than that in extramural environment. Statistically significant difference existed between intramural and extramural environments at all the sites, with maximum difference at a site, which is old crowded area of the city. During the 2-years investigations, 11 species of Penicillium were isolated from the indoor air while nine species were isolated from the air outside the homes. The dominant species of Penicillium in indoor as well as outdoor air were P. citrinum (33.78 and 32.81), P. oxalicum (19.70 and 22.60), and P. chrysogenum (17.64 and 14.50). The percentage of the Penicillium in the indoor air was 10.70 while it was 8.36 in outdoor air. Indoor air showed the presence of P. glaber and P. sclerotiorum, which were absent in the outdoor air.     

Survey of the genus penicillium for tremorgenic toxin production

A preliminary survey of the genusPenicillium reveals that production of a trem-orgenic toxin is confined to certain species of the Subsection Fasciculata, Section Asymmetrica. The most intense toxin synthesis was noted in strains ofP. crustosum, P. cyclopium, P. grannulatum, andP. palitans.

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Zusammenfassung Eine vorläufige Übersicht der GattungPenicillium zeigt, daß die Erzeugung eines tremorgenischen Toxins an gewisse Arten der Subsektion Fasciculata, Section Asymmetrica beschränkt ist. Die kräftigste Toxinsynthesis war inP. crustosum, P. cyclopium P. granulatum, undP. palitans beobachtet.

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This is a laboratory of the Northern Utilization Research and Development Division, Agricultural Research Service, U.S. Department of Agriculture.     

Antifungal and Insecticidal Activities of Selected Plant Species from Cloud Forest of Veracruz,Mexico: A Contribution to the Search of Novel Control Agents against Ambrosia Pest Complexes

The antifungal and insecticidal activities of 34 extracts from 27 plant species were evaluated against fungal phytopathogens of the genus Fusarium and Xyleborus Scolytine ambrosia beetles involved in Fusarium dieback (FD) and laurel wilt (LW) diseases. Sixteen extracts caused mycelial growth inhibition (MGI) above 23 % at 2 mg mL⁻¹ against F. solani, those from S. nudum and M. argyrophylla exhibited the highest MGI (57 % and 49 %, respectively). Thirteen extracts displayed significant antifungal activity against F. kuroshium, those from C. nocturnum and M. argyrophylla exhibited the highest MGI (100 % and 54.9 %, respectively). Additionally, ten plants extracts caused mortality in at least one of the beetle species tested, mainly from Solanaceae species. In the most active species, 39 phenolics were identified that may have contributed to their biological effects. This study is one of the first to report the potential of plant-derived natural products against the causative agents of FD and LW.     

Indoor airbornePenicillium species in Taiwan

Penicillium species, well-known indoor airborne fungi, are considered to be important causative agents of extrinsic bronchial asthma. For selecting the proper species for clinical investigations, it is important to know which are found indoors. In the present study, 176 plates of modified Mehrlich's Medium were exposed for 20 min in living rooms and bedrooms of 88 homes in Taipei, Taiwan. A total of 5897 colonies of fungi were isolated, varying from one to 329 per house. Of the total, 910 colonies (15.4%) were identified asPenicillium spp. The three most frequently encountered species wereP. citrinum, P. crustosum andP. implicatum. Penicillium citrinum accounted for 40.5% (369 from 28 houses);P. implicatum, for 5.2% (47 from three houses); andP. crustosum, for 2.1% (19 from seven houses).Penicillium citrinum was judged to be the most prevalent airborne fungus in Taiwan. The allergenic significance of this mold is under further investigation.     

Antibacterial effects of knotwood extractives on paper mill bacteria

Hydrophilic knotwood extracts from 18 wood species were assessed in disc diffusion and liquid culture tests for antibacterial effects against three species of paper mill bacteria. The Pinus sylvestris, P. resinosa, P. contorta, and P. banksiana extracts decreased or inhibited bacterial growth. The susceptibility order was P. sylvestris > P. resinosa > P. contorta > P. banksiana, correlating with the concentrations of pinosylvin and pinosylvin monomethyl ether in these wood species. Also, Pseudotsuga menziesii and Thuja occidentalis extracts had a small inhibitory effect. The Gram-positive Bacillus coagulans was more susceptible to the extracts than the Gram-negative Burkholderia multivorans and Alcaligenes xylosoxydans. The main components in the Pinus knotwood extracts were pinosylvin monomethyl ether and pinosylvin, suggesting these to be the active components. Therefore, pure pinosylvin, pinosylvin monomethyl ether, and dihydro-pinosylvin monomethyl ether were also tested. All compounds showed antibacterial effects. However, higher concentrations were needed for these pure compounds than for the knotwood extracts. Pinosylvin had stronger antibacterial effects than pinosylvin monomethyl ether. This work shows that knotwood extracts, especially from Pinus species, have a potential for use as natural biocides in papermaking.