Airborne fungal spores in the coastal plain of Israel: A preliminary survey

Airborne spores were monitored during the years 1993–1995 in three cities along the coastal plain of Israel: Ramat Gan, Tel Aviv (Ramat Aviv) and Haifa. Seasonal fluctuations in the concentration of airborne spores were recorded. The following genera of fungi were identified:Alternaria, Cladosporium, Coprinus, Curvularia, Drechslera. Diplococcum, Epicoccum, Fusarium, Leptosphaeria, Pithomyces, Puccinia, Sphacelotheca, Stemphylium andUstilago. Unidentified spores were very rare and in negligible numbers. The dominant airborne fungal spores wereCladosporium andAlternaria. The monthly variations in airborne spores, observed among the three cities, seem to be rather minor. The recorded levels of airborne spores were below the concentrations that are accepted as threshold levels for provocation of clinical responses.     

Fine Structure of Five Species of Myxomycetes with Clustered Spores*

SYNOPSIS. Electronmicroscopic studies were made of 5 species of Myxomycetes with clustered spores to determine the exact mechanism holding the spores together. In Dianema corticatum the neighboring spore walls were closely appressed and appeared to be fused at numerous points. In Trichia synsporum there was a substance between the outer spore walls of adjacent spores which seemed to cement them together. In Badhamia nitens the spore walls were completely fused along their closely appressed surfaces. The adjacent spores of Badhamia versicolor appeared to be free, or if any fusion point was present it was only at the tips of the ornamentation. In Physarum bogoriense such tips were in contact with those of neighboring spores and were occasionally fused at these points.     

Preliminary studies on the effect of the Burkard alternate orifice on airborne fungal spore concentrations

The Burkard 7-day spore trap with standard orifice is commonly used by researchers in sampling outdoor air. The alternate orifice is reported to have higher efficiency in collecting small airborne fungal spores; however, no previous studies compared Burkard samplers with different orifices. This study was conducted to study the effect of the alternate orifice on the concentration of airborne fungal spores. Air samples were collected from July to October 2005 with two Burkard spore traps, one had the standard orifice and the second had the alternate orifice. The two spore traps were located on the roof of a building (12 m height) at the University of Tulsa, Oklahoma. Burkard daily slides were analyzed for airborne spores by light microscopy. The data from the two samplers were statistically analyzed using t-tests. The results indicated that the alternate orifice had significantly higher concentrations of Penicillium/Aspergillus-type spores and basidiospores than the standard orifice. By contrast, the standard orifice had significantly higher concentrations of Alternaria, ascospores, and other spores than the alternate orifice. The alternate orifice can be used to increase the efficiency of trapping small spores, which can be underestimated by using the standard orifice. However, additional comparison in other months of the year is recommended.     

Mechanism of killing of spores of Bacillus cereus and Bacillus megaterium by wet heat

Aims: To determine the mechanism of wet heat killing of spores of Bacillus cereus and Bacillus megaterium. Methods and Results: Bacillus cereus and B. megaterium spores wet heat‐killed 82–99% gave two bands on equilibrium density gradient centrifugation. The lighter band was absent from spores that were not heat‐treated and increased in intensity upon increased heating times. These spores lacked dipicolinic acid (DPA) were not viable, germinated minimally and had much denatured protein. The spores in the denser band had viabilities as low as 2% of starting spores but retained normal DPA levels and most germinated, albeit slowly. However, these largely dead spores outgrew poorly if at all and synthesized little or no ATP following germination. Conclusions: Wet heat treatment appears to kill spores of B. cereus and B. megaterium by denaturing one or more key proteins, as has been suggested for wet heat killing of Bacillus subtilis spores. Significance and Impact of the Study: This work provides further information on the mechanisms of killing of spores of Bacillus species by wet heat, the most common method for spore inactivation.     

The preparation,germination properties and stability of superdormant spores of Bacillus cereus

Aims: To determine yields, germination and stability of superdormant Bacillus cereus spores. Methods and Results: Superdormant B. cereus spores were isolated by germination with high concentrations of inosine or l ‐alanine in 2–5% yield and did not germinate with high concentrations of either of these germinants, but germinated like starting spores with Ca‐DPA, dodecylamine, l ‐alanine plus inosine or concentrated complete medium. Yields of superdormant spores from germinations with low inosine concentrations were higher, and these spores germinated poorly with low inosine, but relatively normally with high inosine. Yields of superdormant spores were also higher when nonheat‐activated spores were germinated. Superdormant spores stored at 4°C slowly recovered some germination capacity, but recovery was slowed significantly at ?20°C and ?80°C. Conclusions: Factors that influence levels of superdormant B. cereus spores and the properties of such spores are similar to those in B. megaterium and B. subtilis, suggesting there are common mechanisms involved in superdormancy of Bacillus spores. Significance: Superdormant spores are a major concern in the food industry, because the presence of such spores precludes decontamination strategies based on triggering spore germination followed by mild killing treatments. Studies of the properties of superdormant spores may suggest ways to eliminate them.     

Acaulosporoid glomeromycotan spores with a germination shield from the 400-million-year-old Rhynie chert

Scutellosporites devonicus from the Early Devonian Rhynie chert is the only fossil glomeromycotan spore taxon known to produce a germination shield. This paper describes a second type of glomeromycotan spore with a germination shield from the Rhynie chert. In contrast to S. devonicus, however, these spores are acaulosporoid and develop laterally in the neck of the sporiferous saccule. Germination shield morphology varies, from plate-like with single or double lobes to tongue-shaped structures usually with infolded margins that are distally fringed or palmate. Spore walls are complex and appear to be constructed of at least three wall groups, the outermost of which includes the remains of the saccule. The complement of features displayed by the fossils suggests a relationship with the extant genera Ambispora, Otospora, Acaulospora or Archaeospora, but which of these is the closest extant relative cannot be determined. The acaulosporoid spores from the Rhynie chert document that this spore type was in existence already ∼400 mya, and thus contribute to a more complete understanding of the evolutionary history of the Glomeromycota. This discovery pushes back the evolutionary origin of all main glomeromycotan groups, revealing that they had evolved before rooted land plants had emerged.     

Otospora bareai, a new fungal species in the Glomeromycetes from a dolomitic shrub land in Sierra de Baza National Park (Granada, Spain)

A new fungal species of the Glomeromycetes was isolated from the rhizosphere of Pterocephalus spathulatus and Thymus granatensis, two rare endemic plants growing on dolomite in the Sierra de Baza (Granada, southern Spain). The fungus was propagated in pot cultures of Sorghum vulgare and Trifolium pratense for 4 y and it is described here on the basis of the spores found in nature and formed in pot cultures. Its brown spores (140-210 microm diam) form laterally on a persistent, brown stalk (=neck) of a sporiferous saccule. They have two walls without ornamentation: a brown, three- to four-layered outer wall and a hyaline two- to three-layered inner wall. The unique combination of spore formation and spore wall structure does not fit with any of the known fungal genera. Spore formation is similar to that of Acaulospora spp. and Archaeospora trappei, but Acaulospora spp. has three spore walls with a characteristic "beaded" wall, and the outer wall of Ar. trappei is simple, thin, hyaline and only bilayered. Spore wall structure of the new fungus is similar to that of Entrophospora infrequens, however this fungus forms its spores internally, inside the hyphal stalk of the sporiferous saccule. Molecular analyses of the small subunit of the ribosomal gene phylogenetically place the new fungus next to Diversispora spurca, which forms one-walled glomoid spores (i.e. terminally on hyphae). Based on these analyses we place the new fungus into a new genus in the family Diversisporaceae under the epithet Otospora bareai.     

The variation in spore concentrations of selected fungal taxa associated with weather conditions in Szczecin,Poland, 2004–2006

The investigation of airborne fungal spore concentrations was carried out in Szczecin, Poland between 2004 and 2006. The objective of the studies was to determine a seasonal variation in concentrations of selected fungal spore types due to meteorological parameters. The presence of spores of ten taxa: Cladosporium, Ganoderma, Alternaria, Epicoccum, Didymella, Torula, Dreschlera‐type, Polythrincium, Stemphylium and Pithomyces was recorded in Szczecin using a volumetric method (Hirst type). Fungal spores were present in the air in large numbers in summer. The highest concentrations were noted in June, July and August. The peak period was recorded in August for most of the studied spore types: Ganoderma, Alternaria, Epicoccum, Dreschlera‐type, Polythrincium and Stemphylium. Cladosporium and Didymella spores reached their highest concentrations in July while concentrations of Torula were highest in May and Pithomyces in September. Multiple regression analysis was performed for three fungal seasons: 2004, 2005, and 2006. Spore concentrations were positively correlated with minimum temperature for seven spore types in 2004, for five spore types in 2005, and for eight spore types in 2006 (significance level of α = 0.05). Some spore types are also significantly correlation among their concentrations, pressure, relative humidity and rain. Minimum temperature appeared to be the most influential factor for most spore types.     

Summer airborne mycoflora of Timişoara (Romania) and relationship to meteorological parameters

The present aeromycological investigation was undertaken to study atmospheric fungal spores in Timi?oara (western Romania). This study was carried out using a Hirst type volumetric sampler. The study revealed the existence of a rich airborne mycoflora. The atmospheric fungal spores were classified and evaluated into three groups (‘major’, ‘minor’ and ‘sporadic’) depending upon their catch percentage in the air. Cladosporium/Fusarium/Leptosphaeria-group, Alternaria, Helminthosporium airborne fungal spores and airborne fungal fragments regularly recorded (frequency 100% of days). Cladosporium accounted for 81.09% of the outdoor fungal spores. The airborne fungal fragments have been identified as abundant in our geographic area. Spearman’s correlations were applied to meteorological parameters and airborne fungal spore concentrations. In addition, correlations were calculated between the fungal spore concentrations and the meteorological variables from the previous day. A total of eleven weather factors were selected for this investigation. Following Spearman’s correlations, I identified two patterns of behaviour: most of the airborne fungal spores prefer cloudiness, lower near-surface soil temperature, lower atmospheric pressure, higher relative humidity and precipitation (pattern A) while other spore concentrations favour increased sunshine, higher near-surface soil temperature and dry conditions (pattern B). The behaviour of some fungal spores during the warm season has proven unclear (pattern C). This study demonstrates the need for investigations throughout the year and the evaluation with complementary statistical methods, regarding the correct interpretation of airborne mycoflora relationships with meteorological parameters.     

Circadian periodicity of airborne pollen and spores; significance of sampling height

Summary The circadian periodicity of the five allergologically most important pollen types in S. Finland and two different spore types were studied at two sites, about 200 meters apart: one Burkard sampler situated on a roof at a height of 15 meters and the other on a garden lawn at ground level. The circadian rhythms of pollen released close to the ground (herb pollen) did not correlate at the two sampling heights used. Variations in pollen from high sources (trees) were significantly correlated at the two heights used. The circadian rhythms of spores from low source distant from the sampling sites (Suillus as an example of forest fungi) were not always correlated; instead, spores with sources at different heights (Cladosporium as an example) had similar circadian periodicity not depending on the sampling level. It was shown that, when studying circadian rhythms of atmospheric particles, the sampling height used is often of great importance. Circadian rhythms of the same pollen and spore types for ten years at the higher sampling site are presented for long-term comparison.     

Sporicidal activity of tertiary butyl hydroperoxide

Tertiary butyl hydroperoxide (t-BOOH) was found to be sporicidal for Bacillus megaterium ATCC19213. Sporicidal action was very temperature dependent, and the potency of t-BOOH increased about tenfold for each increase in temperature of 15 °C over the range from 30° to 70 °C. At still higher temperatures, heat and molar levels of t-BOOH were mutually potentiating for killing. Vegetative cells and germinated spores were some thousand times less resistant to t-BOOH than dormant spores. The order of resistance for spores was: Bacillus stearothermophilus ATCC7953 > Bacillus subtilis var. niger = Bacillus megaterium ATCC33729 > Bacillus megaterium ATCC19213. Killing was not enhanced by decoating and occurred without germination or loss of refractility of the spores. Spore resistance to t-BOOH was lower at more acid pH values and was decreased also by demineralization. Spores could be protected by the chelator o-phenanthroline, especially in association with Fe²⁺. Overall, it seemed that killing was associated with nonmetabolic formation of alkyl peroxyl radicals, which are thought to be responsible for killing of vegetative cells by organic hydroperoxides.Abbreviation A-BOOH tertiary butyl hydroperoxide     

A new late Westphalian fossil marattialean fern from Nova Scotia

Sydneia manleyi gen. et sp. nov. is based on part of a fertile frond from the upper Westphalian D of the Sydney Coalfield, Nova Scotia, Canada. It has small synangia composed of laterally fused sporangia that are elongate and with a circular cross-section. The sporangia yielded variably sized monolete and trilete spores with laevigate and microspinate ornamentation; intermediate forms were also observed. The spores can be correlated with the sporae dispersae species Latosporites minutus , Punctatosporites oculus and Laevigatosporites minimus . Size distribution of the spores is variable and highly skewed, suggesting heterogeneity of the spores within the sporangium. Spore ultrastructure indicates that the fossil is part of a fern, and the morphology of the spores and synangia indicate marattialean affinities.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 199–212.     

Effects of media composition on submerged culture spores of the entomopathogenic fungus,Metarhizium anisopliae var. acridum Part 2: Effects of media osmolality on cell wall characteristics,carbohydrate concentrations,drying stability,and pathogenicity

This study evaluates osmolality of a submerged conidia-producing medium in relation to the following spore characteristics: yield, morphology (dimensions and cell wall structure), chemical properties of cell wall surfaces (charge, hydrophobicity, and lectin binding), cytoplasmic polyols and trehalose, and performance (drying stability and pathogenicity). Spore production was increased by the addition of up to 150 g l^?1 polyethylene glycol 200 (PEG). Spores from high osmolality medium (HOM spores) containing 100 g l^?1 PEG had thin cell walls and dimensions more similar to blastospores than submerged conidia or aerial conidia. However, a faint electron-dense layer separating primary and secondary HOM spores’ cell walls was discernable by transmission electron microscopy as found in aerial and submerged conidia but not found in blastospores. HOM spores also appeared to have an outer rodlet layer, unlike blastospores, although it was thinner than those observed in submerged conidia. HOM spores’ surfaces possessed hydrophobic microsites, which was further evidence of the presence of a rodlet layer. In addition, HOM spores had concentrations of exposed N-acetyl-β-d-glucosaminyl residues intermediate between blastospores and submerged conidia potentially indicating a masking of underlying cell wall by a rodlet layer. All spore types had exposed α-d-mannosyl and/or α-d-glucosyl residues, but lacked oligosaccharides. Similar to blastospores, HOM spores were less anionic than submerged conida. Although HOM spores had thin cell walls, they were more stable to drying than blastospores and submerged conidia. Relative drying stability did not appear to be the result of differences in polyol or trehalose concentrations, since trehalose concentrations were lower in HOM spores than submerged conidia and polyol concentrations were similar between the two spore types. HOM spores had faster germination rates than submerged conidia, similar to blastospores, and they were more pathogenic to Schistocerca americana than submerged conidia and aerial conidia.     

Influence of urban climate upon distribution of airborne Deuteromycete spore concentrations in Mexico City

 The effect of an urban climate upon the spatial and temporal distribution of Deuteromycete spores was studied during 1991 using Burkard volumetric spore traps in two areas of Mexico City with different degrees of urbanization. Deuteromycete conidia formed the largest component of the total airborne fungal spore load in the atmosphere of Mexico City, contributing 52% of the spores trapped in an urban-residential area (southern area) and 65% of those in an urban-commercial area (central area). Among the most common spore types, Cladosporium and Alternaria showed a marked seasonal periodicity with significant differences in concentration (P<0.05) between the dry and wet seasons. Maximum conidial concentrations were found during the end of the wet season and the beginning of the cool, dry season (October–December). Daily mean concentrations of the predominant airborne spore types did not differ significantly between the southern and central areas. Daily mean spore concentrations were significantly correlated (P<0.05) in southern and central areas with maximum temperature (south, r = –0.35; central, r = –0.40) and relative humidity (south, r = 0.43; central, r = 0.29) from the previous day. Moreover, multiple regression analysis of spore concentrations with several meteorological factors showed significant interactions between fungal spores, relative humidity and maximum temperature in both areas. The diurnal periodicity of Cladosporium conidia characteristically showed two or three peaks in concentration during the day at 0200–0400, ∼ 1400 and 2000–2200 hours, while that of Alternaria showed only one peak (1200 to 2000 hours) in both areas. Maximum concentrations of these spores generally occurred 2–4 h earlier in the southern than in the central area. The lag in reaching maximum concentrations in the central area probably resulted from differences in the local conditions between the study areas, and from spores transported aerially into the city from distant sources. The analysis of maximum hourly concentrations of Cladosporium and Alternaria spores during 1 month of the dry season (February), and another month of the wet season (September) showed significant differences between the two study areas. Environmental factors and sources (green areas) affected diurnal changes in conidial concentration in the southern area (urbanization index, UI, 0.25), but not in the central area (UI 0.97). In general, spore concentrations were greatest in the southern area when relative humidities were low, and temperatures and wind velocities were high. It was difficult to establish effects of climatic factors on the spore concentration in the city centre. This probably results from the large amounts of air pollution, the heat island phenomenon, and from the distant origin of trapped conidia obviating aerial transport. Nevertheless, the seasonal and diurnal distributions of conidia found were similar to those reported for other tropical regions of the world. Received: 13 August 1996 / Accepted: 4 December 1996     

Early Development and Tissue Distribution of Pseudoloma neurophilia in the Zebrafish,Danio rerio

The early proliferative stages of the microsporidian parasite, Pseudoloma neurophilia were visualized in larval zebrafish, Danio rerio, using histological sections with a combination of an in situ hybridization probe specific to the P. neurophilia small‐subunit ribosomal RNA gene, standard hematoxylin‐eosin stain, and the Luna stain to visualize spores. Beginning at 5 d post fertilization, fish were exposed to P. neurophilia and examined at 12, 24, 36, 48, 72, 96, and 120 h post exposure (hpe). At 12 hpe, intact spores in the intestinal lumen and proliferative stages developing in the epithelial cells of the anterior intestine and the pharynx and within hepatocytes were observed. Proliferative stages were visualized in the pancreas and kidney at 36–48 hpe and in the spinal cord, eye, and skeletal muscle beginning at 72 hpe. The first spore stages of P. neurophilia were observed at 96 hpe in the pharyngeal epithelium, liver, spinal cord, and skeletal muscle. The parasite was only observed in the brain of larval fish at 120 hpe. The distribution of the early stages of P. neurophilia and the lack of mature spores until 96 hpe indicates that the parasite gains access to organs distant from the initial site of entry, likely by penetrating the intestinal wall with the polar tube.     

One-Year pollen and spore calendars of Saudi Arabia Al-Khobar, Abha and Hofuf

Airborne Pollen grains and Spores of three different cities viz., Al-Khobar (1987–1988), Abha (1991–1992) and Hofuf (1992–1993) in Saudi Arabia were studied using Burkard Volumetric Seven-Day Spore Trap. The data were analyzed in relation to their allergenic capability and one-year pollen and spore calendars were designed to correlate the patients’ symptoms as well as for selection of appropriate allergen extracts for diagnosis and treatment of allergic diseases. Amongst pollen group, Amaranthus viridis, Plantago spp., Chenopodium album, Ricinus communis, Rumex vesicarius, Juniperus spp., Parkinsonia aculeata, Prosopis spp., and Phoenix dactylifera were some of the frequent types. Amongst the fungal spores group Cladosporium, Smuts spores, Colored basidiospores, Alternaria, Ulocladium and Drechslera were the dominant types.     

Analysis of the slow germination of multiple individual superdormant Bacillus subtilis spores using multifocus Raman microspectroscopy and differential interference contrast microscopy

Aim: To analyse the dynamic germination of hundreds of individual superdormant (SD) Bacillus subtilis spores. Methods and Results: Germination of hundreds of individual SD B. subtilis spores with various germinants and under different conditions was followed by multifocus Raman microspectroscopy and differential interference contrast microscopy for 12 h and with temporal resolutions of ≤30 s. SD spores germinated poorly with the nutrient germinant used to isolate them and with alternate germinants targeting the germinant receptor (GR) used originally. The mean times following mixing of spores and nutrient germinants to initiate and complete fast release of Ca‐dipicolinic acid (CaDPA) (T_lag and T_release times, respectively) of SD spores were much longer than those of dormant spores. However, the ΔT_release times (T_release?T_lag) of SD spores were essentially identical to those of dormant spores. SD spores germinated almost as well as dormant spores with nutrient germinants targeting GRs different from the one used to isolate the SD spores and with CaDPA that does not trigger spore germination via GRs. Conclusions: Since (i) ΔT_release times were essentially identical in GR‐dependent germination of SD and dormant spores; (ii) rates of GR‐independent germination of SD and dormant spores were identical; (iii) large increases in T_lag times were the major difference in the GR‐dependent germination of SD as compared with spores; and (iv) higher GR levels are correlated with shorter T_lag times, these results are consistent with the hypothesis that low levels of a GR are the major reason that some spores in a population are SD with germinants targeting this same GR. Significance and Impact of the Study: This study provides information on the dynamic germination of individual SD spores and improves the understanding of spore superdormancy.     

Histidine acts as a co‐germinant with glycine and taurocholate for Clostridium difficile spores

Aims: It is well established that the bile salt sodium taurocholate acts as a germinant for Clostridium difficile spores and the amino acid glycine acts as a co‐germinant. The aim of this study was to determine whether any other amino acids act as co‐germinants. Methods and Results: Clostridium difficile spore suspensions were exposed to different germinant solutions comprising taurocholate, glycine and an additional amino acid for 1 h before heating shocking (to kill germinating cells) or chilling on ice. Samples were then re‐germinated and cultured to recover remaining viable cells. Only five amino acids out of the 19 common amino acids tested (valine, aspartic acid, arginine, histidine and serine) demonstrated co‐germination activity with taurocholate and glycine. Of these, only histidine produced high levels of germination (97·9–99·9%) consistently in four strains of Cl. difficile spores. Some variation in the level of germination produced was observed between different PCR ribotypes, and the optimum concentration of amino acids with taurocholate for the germination of Cl. difficile NCTC 11204 spores was 10–100 mmol l^?1. Conclusions: Histidine was found to be a co‐germinant for Cl. difficile spores when combined with glycine and taurocholate. Significance and Impact of the Study: The findings of this study enhance current knowledge regarding agents required for germination of Cl. difficile spores which may be utilized in the development of novel applications to prevent the spread of Cl. difficile infection.     

Indoor air quality in schools: exposure to fungal allergens

This study examined indoor air quality within schools in Kansas City, Spokane, Santa Fe, and Orlando. Air sampling was undertaken with both Andersen Single Stage Samplers and Burkard Personal Air Samplers. The data show a wide range of indoor exposures ranging from less than 100 colony forming units (CFU/m³) for viable fungi and 100 spores/m³ for total spores in Spokane and Santa Fe to concentrations over 6000 CFU/m³ for viable fungi and 15 000 spores/m³ for total fungi in Orlando and Kansas City, respectively. In the majority of sites the indoor airspora reflected the outdoor taxa withCladosporium the most abundant genus identified; however, several indoor locations had elevated levels ofPenicillium andAspergillus indicating possible sources of indoor contamination. Airborne basidiospores and smut spores were also fairly abundant in the schools and were among the top five taxa identified. The data also indicated that the airborne concentrations vary significantly during the day and between classrooms within each school. Continued studies in schools are needed to fully assess both the exposure levels and the clinical significance to atopic children allergic to these spores.     

Aeromycological study in the Cathedral of Santiago de Compostela (Spain)

A study of airborne fungi was carried out in the architectural complex of the Cathedral of Santiago de Compostela (Spain) during 2002, by using viable volumetric sampling methods. This resulted in a total of 35 identified taxa, of which the most abundant were: Alternaria, Aspergillus, Cladosporium and Penicillium. Sampling was completed with data from the outdoor atmosphere and swab samples in specific places.In general there were no statistically significant indoor/outdoor differences and in both cases the highest CFU m-3 were obtained during the spring-summer. Similar relatively low numbers of the same fungi were likewise detected at different points in the Cathedral nave, while up to nearly 6500 CFU m-3 were recorded in the Corticela Chapel. The study of intradiurnal levels carried out in the Cathedral nave reveals greater abundance of fungal concentrations at 13:00 h, the moment of massive influx of visitors in the Cathedral, with 406 CFU m-3 compared to the 380 CFU m-3 sampled at 9:00 h and the 350 CFU m-3 at 21:00 h. The whole investigation is the first study of the atmospheric fungal content of the Cathedral of Santiago de Compostela.