Experimental toxicoinfection in infant mice challenged with spores of Clostridium botulinum type E

Conventionally raised suckling mice were given 10(7) spores of a strain of Clostridium botulinum type E. Most but not all infant mice aged 8 through 19 days at the time of administration died after developing symptoms typical of botulism. However, none of the infant mice challenged with the spores at dose levels lower than 10(6) spores/mouse developed illness.     

QUALITATIVE AND QUANTITATIVE STUDIES OF THE SWIMMING BEHAVIOR OF HINCKSIA IRREGULARIS SPORES (PHAEOPHYCEAE)

The swimming behavior of spores of the brown alga Hincksia irregularis was analyzed using computer-assisted motion analysis. We distinguished five main swimming patterns: straight paths, search circles, orientation, gyration, and wobbling. We suggest different functional values for the individual swimming patterns. Straight paths, search circles, and orientation are different but all may be important in small-scale movements in the benthic boundary layer. As such, they could enable a spore to find a suitable microenvironment for germination and growth. Gyration occurs during the initial reversible phase of adhesion that can lead to settlement. Wobbling is typical for irritated or mechanically damaged spores and does not seem to be a typical pattern associated to settlement. The dominant swimming patterns changed with spore age (10 ± 5 to 60 ± 5 min of spore age), with young spores mainly swimming in straight paths and search circles and older spores in orientation and gyration. This change in swimming patterns can be quantified by speed (decrease over time) and rate of change of direction (increase over time). Based on these results, we suggest that computer-assisted motion analysis of the swimming behavior of H. irregularis spores can be used to develop bioassays with both ecological and technological relevance.     

同步辐射软X射线对枯草杆菌的诱变效应

采用同步辐射软X射线对枯草芽孢杆菌(Bacillus subtilis)1831菌株进行辐照处理,研究了不同剂量下3．1nm的软X射线对其芽孢的失活和诱变作用。结果表明：同步辐射软X射线对枯草杆菌芽孢的剂量存活曲线表现为典型的“肩型”,对芽孢的失活作用属于“单靶多击”方式,失活击中数等于3。根据脱脂牛奶平板上蛋白酶活力大小的测量统计,以变异系数作为诱变效应指标,软X射线对芽孢具有一定的诱变作用。     

Food utilization values of gypsy moth Lymantria dispar (Lepidoptera: Lymantriidae) larvae infected with the microsporidium Vairimorpha sp. (Microsporidia: Burenellidae)

Infection of the gypsy moth, Lymantria dispar, with the microsporidium Vairimorpha sp. strongly influences the development of the host in ways typical of many species of terrestrial entomopathogenic Microsporidia; growth is reduced while development time is extended in infected insects. The appearance of the different stages of the parasite in the host relative to the elapsed time after oral infection, as well as the influence of the parasite proliferation on food utilization of the host, were examined. At 3 days postinfection, midgut muscle cells were infected with primary spores, and the fat body tissues contained meronts, sporonts, and primary spores. Many more fat body cells contained vegetative stages and primary spores at 4 and 5 days postinfection, and diplokaryotic spores and immature octospores were also present. Approximate digestibility of infected larvae increased during this time period, whereas the conversion of ingested and digested food to body substance decreased. The relative growth rate of infected and uninfected groups did not differ significantly between 4 and 5 days postinfection, although the relative consumption rate in infected L. dispar larvae was higher. Between 8 and 10 days postinfection, the relative growth rate of uninfected larvae increased. The infected group did not demonstrate this increase at a time period characterized by maturation of diplokaryotic spores and octospores in larval fat body tissues. Total body weight of uninfected larvae remained higher than that of infected larvae after 8 days postinfection.     

Immunobiological properties of 100 years old Bacillus anthracis vaccine strains Lange

The results of the study of B. anthracis vaccine strains Lange 1 and 2 in the form of spores after prolonged storage in 30% glycerine solution at room temperature are presented. The study revealed that spores stored for 100 years germinated and bacterial cells proved to be viable when cultivated in artificial nutrient media and in vivo. They exhibited typical cultural, morphological, biochemical, virulent, antigenic and immunogenic properties.     

Growth and sporulation of the arbuscular mycorrhizal fungus Glomus caledonium in dual culture with transformed carrot roots

Glomus caledonium was established in a dual culture with Ri T-DNA-transformed carrot roots. A modification of the minimal M medium buffered at pH 6.50 with 10 mM MES and solidified with 0.4% unpurified gellan gum allowed spore germination and formation of the symbiosis, together with the development of an extensive extramatrical mycelium and sporulation. Spore production increased with culture generation and most spores were viable. These spores colonized carrot roots and completed the fungal life cycle. In many cultures, sporulation was accompanied by the formation of arbuscule-like structures on short and thickened lateral branches of main hyphae. Root colonization was of the Paris-type with hyphae spreading intracellularly. Most colonized root cells contained coils of thickened hyphae, sometimes surrounded by fine hyphae, but no typical arbuscules were observed. Accepted: 26 January 2000     

Fungal spores: dormancy, germination, chemical composition, and role in biotechnology (review)

This review is focused on one of the stages of ontogenesis distinctive by its particular tolerance to the action of unfavorable factors and ability to retain the genomic material for a long period of time, i.e., fungal spores. The major part is devoted to the characterization of the specific stage typical for spores, which is called dormancy. Data are presented characterizing the carbohydrate and lipid composition of spores, with special attention being paid to the role of carbohydrate protectors, in particular, trehalose and mannite, as well as to the role of rafts in the process of sporogenesis. The role of special compounds called autoinhibitors and autostimulators in the process of exit from dormancy is discussed. The final section deals with the role of spore seeding material in biotechnological processes. Data on the correlation between the chemical composition of spores, their ability to remain dormant, and the germination process are considered. Special biotechnological approaches are presented for the first; they allow for the preservation of the germinating ability of spores, intensification of sporogenesis, changes in the ratio of final fermentation products, and an increase in their yield.     

Fungal spores: Dormancy, germination, chemical composition, and role in biotechnology (review)

This review is focused on one of the stages of ontogenesis distinctive by its particular tolerance to the action of unfavorable factors and ability to retain the genomic material for a long period of time, i.e., fungal spores. The major part is devoted to the characterization of the specific stage typical for spores, which is called dormancy. Data are presented characterizing the carbohydrate and lipid composition of spores, with special attention being paid to the role of carbohydrate protectors, in particular, trehalose and mannite, as well as to the role of rafts in the process of sporogenesis. The role of special compounds called autoinhibitors and autostimulators in the process of exit from dormancy is discussed. The final section deals with the role of spore seeding material in biotechnological processes. Data on the correlation between the chemical composition of spores, their ability to remain dormant, and the germination process are considered. Special biotechnological approaches are presented for the first; they allow for the preservation of the germinating ability of spores, intensification of sporogenesis, changes in the ratio of final fermentation products, and an increase in their yield.     

A Novel Phytomyxean Parasite Associated with Galls on the Bull-Kelp Durvillaea antarctica (Chamisso) Hariot

Durvillaea antarctica (Fucales, Phaeophyceae) is a large kelp of high ecological and economic significance in the Southern Hemisphere. In natural beds along the central coast of Chile (Pacific Ocean), abnormal growth characterized by evident gall development and discolorations of the fronds/thallus was observed. Analysing these galls by light microscopy and scanning electron microscopy revealed the presence of endophytic eukaryotes showing typical characteristics for phytomyxean parasites. The parasite developed within enlarged cells of the subcortical tissue of the host. Multinucleate plasmodia developed into many, single resting spores. The affiliation of this parasite to the Phytomyxea (Rhizaria) was supported by 18S rDNA data, placing it within the Phagomyxida. Similar microorganisms were already reported once 23 years ago, indicating that these parasites are persistent and widespread in D. antarctica beds for long times. The symptoms caused by this parasite are discussed along with the ecological and economic consequences. Phytomyxean parasites may play an important role in the marine ecosystem, but they remain understudied in this environment. Our results demonstrate for the first time the presence of resting spores in Phagomyxida, an order in which resting spores were thought to be absent making this the first record of a phagomyxean parasite with a complete life cycle so far, challenging the existing taxonomic concepts within the Phytomyxea. The importance of the here described resting spores for the survival and ecology of the phagomyxid parasite will be discussed together with the impact this parasite may have on ‘the strongest seaweed of the world’, which is an important habitat forming and economic resource from the Southern Hemisphere.     

Effect of Bacillus anthracis lethal toxin on human peripheral blood mononuclear cells

Lethal toxin (LeTx) plays a central role in anthrax pathogenesis, however a cytotoxicity of LeTx has been difficult to demonstrate in vitro. No cytolytic effect has been reported for human cells, in contrast to murine cell lines, indicating that cell lysis can not be considered as a marker of LeTx activity. We have recently shown that murine macrophage-like RAW 264.7 cells treated with LeTx or infected with anthrax spores underwent changes typical of apoptotic death. Here we demonstrate that cells from human peripheral blood display a proapoptotic behavior similar to murine cells. TUNEL assay detected a nucleosomal degradation typical of apoptosis in peripheral blood mononuclear cells (PBMC) treated with LeTx. Membrane staining with apoptotic dyes was detected in macrophages derived from monocytes in presence of LeTx. The toxin inhibited production of proinflammatory cytokines in PBMC stimulated with a preparation of Bacillus anthracis cell wall. Infection of PBMC with anthrax spores led to the appearance of a large population of cells stained positively for apoptosis, with a reduced capacity to eliminate spores and vegetative bacteria. The aminopeptidase inhibitor, bestatin, capable of protecting cells from LeTx, restored a bactericidal activity of infected cells. These findings may be explained by LeTx expression within phagocytes and support an important role of LeTx as an early intracellular virulence factor contributing to bacterial dissemination and disease progression.     

Ultrastructure of Bacillus pulvifaciens

The ultrastructure of vegetative cells and spores of Bacillus pulvifaciens was studied by CTEM and SEM methods. The vegetative cells are rods, 1.6–4.5 m long and 0.4–0.6 m wide, exhibiting typical ultrastructural features of Gram-positive bacteria. The spores are of ellipsoidal shape, 0.6×1.2 m in size, with six longitudinal ribs reaching up to 130 nm in height. There are satelite ribs on both sides of the longitudinal ribs, reaching up to 20 nm in height. Between the longitudinal ribs, additional transversal ribs were observed in SEM. A special tubular layer, separating the outer and inner coat of the spores, was revealed in ultrathin sections. This layer seems to be a typical ultrastructural feature of Bacillus pulvifaciens spores.     

Lateral fungal spore movement inside a simulated wall

Moisture inside walls can facilitate mold growth if left untreated. Once spores become airborne they may interact with pressures inside walls. Two laboratory experiments were conducted to determine if airborne spores have the potential to migrate laterally inside walls with and without wiring installations. A simulated wall was fabricated, and Penicillium chrysogenum spores were aerosolized into a distant stud bay and an adjacent stud bay. The wall was subjected to a typical indoor pressure. Spore levels inside the bays were sampled, and a total of 36 trials (n = 36) were conducted. Results of Kruskal–Wallis tests revealed that spore levels inside the sampling bay and the distant bay with wiring installations were not significantly different. Spore levels inside the sampling bay were significantly lower than the adjacent bay without wiring installations (P < 0.05). The findings of the study suggest airborne fungal spores have the potential to move laterally inside walls.     

SPORE ULTRASTRUCTURE OF THE MYXOMYCETE PHYSARUM POLYCEPHALUM

Spores of the true slime mold Physarum polycephalum were examined at several stages of their development by means of scanning and transmission electron microscopy. The spores were globose, spine-covered structures produced within a sporangium enclosed in a tough, noncellular peridium. Cytologically, the spore represented a typical eukaryotic cell, having discrete organelles similar to spores of other myxomycetes. The presence of dictyosomes, helical filaments, and microbodies in these cells, as well as the further elucidation of the cell wall and the “polysaccharide-containing” areas, represent new contributions to the ultrastructure of the myxomycete spore. Of special interest were observations of metaphase nuclei just prior to spore cleavage, interphase nuclei in young spores, and nuclei in mature spores containing synaptonemal complexes. These observations indicate that in Physarum polycephalum mitosis occurs just prior to spore cleavage, and meiosis takes place after spore cleavage.     

Ventilation required to entrain small particles from leaves

Particles are blown from leaves when the wind at the height of the particles exceeds a minimum which is about 5 m/sec for some fungal spores. In the moderate winds typical within a canopy of leaves, the minimum is attained at spore height during brief changes in wind or puffs before the boundary layer grows to particle height. The requisite change in speed to remove spores occurs over a sizeable area only when the speed changes abruptly in a short distance in the direction of the wind.     

Rupture of bacterial spores with ammonium bicarbonate subsequently removed by sublimation

A new method for dry rupture of bacterial spores was developed. Ammonium bicarbonate crystals, used in place of sodium chloride or glass beads, are vigorously shaken with spores. The ammonium bicarbonate is removed subsequently by sublimation, yielding uncontaminated, disrupted spores. Increased recovery of glucose dehydrogenase and protease activity from Bacillus cereus spores was obtained with the new method.     

Germination of spores of Bacillus subtilis with dodecylamine

AIMS: To determine the properties of Bacillus subtilis spores germinated with the alkylamine dodecylamine, and the mechanism of dodecylamine-induced spore germination. METHODS AND RESULTS: Spores of B. subtilis prepared in liquid medium were germinated efficiently by dodecylamine, while spores prepared on solid medium germinated more poorly with this agent. Dodecylamine germination of spores was accompanied by release of almost all spore dipicolinic acid (DPA), degradation of the spore's peptidoglycan cortex, release of the spore's pool of free adenine nucleotides and the killing of the spores. The dodecylamine-germinated spores did not initiate metabolism, did not degrade their pool of small, acid-soluble spore proteins efficiently and had a significantly lower level of core water than did spores germinated by nutrients. As measured by DPA release, dodecylamine readily induced germination of B. subtilis spores that: (a) were decoated, (b) lacked all the receptors for nutrient germinants, (c) lacked both the lytic enzymes either of which is essential for cortex degradation, or (d) had a cortex that could not be attacked by the spore's cortex-lytic enzymes. The DNA in dodecylamine-germinated wild-type spores was readily stained, while the DNA in dodecylamine-germinated spores of strains that were incapable of spore cortex degradation was not. These latter germinated spores also did not release their pool of free adenine nucleotides. CONCLUSIONS: These results indicate that: (a) the spore preparation method is very important in determining the rate of spore germination with dodecylamine, (b) wild-type spores germinated by dodecylamine progress only part way through the germination process, (c) dodecylamine may trigger spore germination by a novel mechanism involving the activation of neither the spore's nutrient germinant receptors nor the cortex-lytic enzymes, and (d) dodecylamine may trigger spore germination by directly or indirectly activating release of DPA from the spore core, through the opening of channels for DPA in the spore's inner membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide new insight into the mechanism of spore germination with the cationic surfactant dodecylamine, and also into the mechanism of spore germination in general. New knowledge of mechanisms to stimulate spore germination may have applied utility, as germinated spores are much more sensitive to processing treatments than are dormant spores.     

Analysis of Airborne Actinomycete Spores with Fluorogenic Substrates

The reactions between seven fluorogenic substrates and different groups of enzymes, esterases, lipases, phosphatases, and dehydrogenases, were studied in a search for a new method for the detection of actinomycete spores. Fluorescence measurement was chosen as a fast and sensitive method for microbial analysis. The focus of the research was on the spores of important air contaminants: Streptomyces albus and Thermoactinomyces vulgaris. For the measurement of the enzymatic activity, the chosen fluorogenic substrate was added to a mixture of spores and nutrient media, and the resulting fluorescence was measured with a spectrofluorometer. Fluorogenic substrates were found to show enzymatic activities even for dormant spores. Comparison of the enzymatic activities of dormant spores with those of vegetative cells showed similarity of the enzymatic profiles but higher activity for vegetative cells. The increase of enzymatic activity from dormant spores to vegetative cells was not linear but fluctuating. The largest fluctuations were found after 4 to 5 h of incubation. The enzymatic activities of S. albus were 10 to 50 times lower than those of T. vulgaris, except for the dehydrogenase activity, which was seven times higher. These results indicate that analysis with fluorogenic substrates has the potential for becoming a fast and sensitive method for the enumeration and identification of airborne actinomycete spores.     

Airborne Pollen and Fungal Spores in Florist Shops in Worcester and in Bristol, UK: A Potential Problem for Occupational Health

Persistent allergies are common in workers in florist shops but little research has been done on the reasons for this. This paper reports an investigation of occupational exposure of florists to pollen and spores in three florist shops over a 2-week period in the autumn of 2000. In each shop three sampling methods were used: Burkard continuous volumetric samplers, deposition plates and low-tac tape for surface samples, including hands of the florists and leaves of a selection of the plants. The florists kept a record of the type of work they undertook each day and of the stock amounts of flowers in the shops. The volumetric traps collected 80 pollen and spore taxa. The average concentrations recorded through the working day were generally low but short-term peak (one hour mean) concentrations of some types were found to be relatively high or very high. For a few taxa these concentrations equalled or exceeded those typical for short-term peaks in wind dispersed pollen types in the ambient air. Fungal spore concentrations of several known allergenic types were also very high for peak periods. This was most notable for Aspergillus spp. which reached extremely high concentrations in one shop, compared with typical peak concentrations in the ambient atmosphere. Low-tac tape leaf samples demonstrated that the flowers' foliage is a major source of the fungal spores. Few pollen grains or fungal spores were found on the hands.     

Purine Auxotrophic Mutants with Altered Spore Color in Streptomyces azureus. ATCC 14921

A purine auxotroph with pale purple spores and a derivative with yellowish orange spores were obtained from the wild-type strain of Streptomyces azureus ATCC 14921, which has bluish green spores. The changed color or pigmentation in the mutants was limited to the spores. They accumulated AIR (5′-phosphoribosyl-5-aminoimidazole) due to the lack of AIR carboxylase activity.     

Lipid composition of cells of heterothallic strains in the developmental cycle of Blakeslea trispora

Lipid compositions in mycelium and spores of Blakeslea trispora heterothallic strains were studied. Distinctions between the strains in the ability to synthesize linolenic acid and in optimal growth temperature were demonstrated. The (-) strain grew at a higher temperature and was unable to synthesize linolenic acid, whereas the (+) strain accumulated this acid up to 20% of total fatty acids. The distinctions between the strains remained at different developmental stages (mycelium and spores). A higher thermophilicity of the (-) strain correlated with a high sterol content, which is typical of thermophilic fungi. The lipid compositions of heterothallic strains studied differed in lipid content, their fractional composition, the degree of unsaturation, and carotenoid composition.