• The Korean Journal of Mycology
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• v.22 no.2
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• pp.138-144
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• 1994

Conditions for isolation of protoplasts from spores and mycelia of Rhizopus nigricans were studied. Larger amount of protoplasts was obtained from swollen spores in liquid medium contained with 5% of 2-deoxy-D-glucose for 4 hours than from mycelia. Enzyme mixture of Novozym 234(2%) and ${\beta}-glucuronidase(5000\;unit/ml)$ was most effective for the isolation of protoplasts from swollen spores and from mycelia. The solution of 0.6 M $MgSO_4$ or mannitol and pH 6.0 showed good results as the osmotic stabilizer and the optimal condition of pH of the enzyme solution for the isolation of protoplast from the swollen spores, respectively. At this condition, $8.0{\times}10^6\;cells/ml$ of protoplasts was obtained from swollen spores by digestion with lytic enzyme mixture for 2 hours.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.616-623
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• 2007

The effects of initial concentration and pulsed pressurization on the inactivation of Clostridium sporogenes spores suspended in deionized water were determined during thermal processing $(TP;\;105^{\circ}C,\;0.1MPa)$ and pressure-assisted thermal processing $(PATP;\;105^{\circ}C\;and\;700MPa)$ treatments for 40 min and 5min holding times, respectively. Different inoculum levels $(10^4,\;10^6\;and\;10^8CFU/ml)$ of C. sporogenes spores suspended in deionized water were treated at $105^{\circ}C$ under 700MPa with single, double, and triple pulses. Thermally treated samples served as control. No statistical significances (p>0.05) were observed among all different inoculum levels during the thermal treatment, whereas the inactivation rates $(k_1\;and\;k_2)$ were decreased with increasing the initial concentrations of C. sporogenes spores during the PATP treatments. Double- and triple-pulsed pressurization reduced more effectively the number of C. sporogenes spores than single-pulse pressurization. The study shows that the spore clumps formed during the PATP may lead to an increase in pressure-thermal resistance, and multiple-pulsed pressurization can be more effective in inactivating bacterial spores. The results provide an interesting insight on the spore inactivation mechanisms with regard to inoculum level and pulsed pressurization.

• The Plant Pathology Journal
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• v.34 no.4
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• pp.335-340
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• 2018

To investigate the difference in the disinfectant efficiency of ozone microbubbles ($O_3MB$) and ozone millibubbles ($O_3MMB$), the morphological change of the treated Fusarium oxysporum f. sp. melonis spores was observed with scanning and transmission electron microscopies (SEM and TEM). The disinfectant efficiency of $O_3MB$ on F. oxysporum f. sp. melonis spores was greater than that of $O_3MMB$. On observation with SEM, it was revealed that morphological change of F. oxysporum f. sp. melonis spores was caused by $O_3MB$ and $O_3MMB$, and damage to the spore surfaces by $O_3MB$ occurred sooner than that by $O_3MMB$. On observation with TEM, it was furthermore confirmed that F. oxysporum f. sp. melonis spores treated with $O_3MB$ induced wavy deformation of cell membrane and the intracellular change different from that with $O_3MMB$. Therefore, the greater disinfection efficiency of $O_3MB$ was suggested to be caused due to the function of the MB in addition to the oxidative power of $O_3$.

• Mycobiology
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• v.47 no.3
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• pp.329-334
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• 2019

Streptomyces blastmyceticus strain 12-6 was isolated from a forest soil sample of Cheonan area on the basis of strong antifungal activities against plant pathogenic fungi. Butanol extracts of the cultural filtrates were active against C. acutatum, C. coccodes, C. gloeosporioides, F. oxysporum, and T. roseum. Active fractions were prepared by thin layer chromatography using silica gel plate; 12-6-2 ($R_f$ 0.36), 12-6-3 ($R_f$ 0.44). Scanning electron microscopy showed that the active fractions caused a change in surface texture of fungal spores from smooth surface to wrinkled surface. The lethal effect on the spores of the active fractions varied from 56% to 100%. It was shown that the spores of C. acutatum were more sensitive to the antifungal fractions than the spores of F. oxysporum. Fluorescence staining using TOTO-1 indicated that the antifungal fractions could make the spores more sensitive to the fluorescence dye. Thus, it was suggested that antifungal agents prepared in this study exhibited the antifungal activity by damaging the plasma membrane of both fungal spores and hyphae. Identification of antifungal agents in the active fraction using GC-MS analysis revealed the presence of cyclo-(Leu-Pro) and 9-octadecenamide as major components that have already been known as antifungal substances.

• Korean Journal of Veterinary Research
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• v.16 no.1
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• pp.11-25
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• 1976

Studies on pathogenicities and developmental stages of Nosema apis (Zander, 1909) were carried out through artificial infection to Nosema free honey bees with various levels of spores isolated from local honey bee colony. The results obtained were summarized as follows: 1. The clinical symptoms were observed as dysentery, enteritis of mid-gut (enlargement and decoloration), crawling posture and shortening of the longevity of worker bees in the rearing honey bee colony inoculated with the spores. 2. Number of spores harvested from laboratory rearing honey bees were progresively increased to 4 weeks after inoculation. The regression equations and coefficients of correlations to various spore levels were as follows in each treatment colony. Colony 1. ($$1,000{\times}10^4spores/ml$$) $$y_{c1}=471{\times}10^{4}x+454{\times}10^4(r=0.65^*$$) Colony 2. ($$500{\times}10^4spores/ml$$) $$y_{c2}=340{\times}10^{4}x+207.8{\times}10^4(r=0.99^{**}$$) Colony 3. ($$100{\times}10^4spores/ml$$) $$y_{c3}=150{\times}10^{4}x+84.2{\times}10^4(r=0.99^{**}$$) Colony 4. ($$10{\times}10^4spores/ml$$) $$y_{c4}=13.8{\times}10^{4}x+13{\times}10^4(r=0.98^{**}$$) 3. Average longevity of worker bees artificially infected with Nosema apis was shortened as 21.7~43.8% compare to the control. (p<.05, p<.01) 4. The spores which were isolated from honey bee colony infected with Nosema disease were ovoid or spherical form, and measured, as a rule, from $4.7{\mu}m$ to $6.1{\mu}m$ (mean $5.3{\mu}m$) in length and from $2.4{\mu}m$ to $3.2{\mu}m$ (mean $2.9{\mu}m$) in width. 5. In the mid-gut of honey bees, the spore was progresively germinated and became trophozoite stage. The trophozoites were grown to meronts and their binary fission were begun. The divided two sporoblasts were developed to the spores which had elastic membrane. The new spores were shed in excreta of honey bees 10~15 day after inoculation at $25{\pm}2$ centigrade. 6. The ultrastructure of spore membrane consisted of three layers, such as, outer, middle and inner layer. The sporoplasm consisting lamellar structure occupied only anterior part of the spore and was often extended to posterior direction where definite vacuoles and a polar filament was able to detect.

• Journal of Sericultural and Entomological Science
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• v.26 no.2
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• pp.1-6
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• 1984

The present study was made to establish the purification method of mature microsporidian spores by iso-density equilibrium technique using percoll and the serological discrimination by an indirect fluorescent antibody technique. The purification of new microsporidian spores took effect in the three steps purification method(precentrifugation-percoll iso-density equilibrium centrifugation-rising). There were clear differences in the size of spores between the new microsporidia and N. bombycis. The spores of N. bombycis is short elliptical of 2.07 in a ratio of length to width in diameter while that of new microsporidia is characterized with long elliptical shape which shows a ratio of 2.76 in length to width in diameter. The specific antigens of new microsporidia K79 spores was showed in the spores wall by the indirect fluorescent antibody reaction, and it was affected by the antibody against N. bombycis which antiserum was diluted in 1:20. It means that the new microsporidia K79 is serologically not identical to N. bombycis.

• The Korean Journal of Mycology
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• v.46 no.4
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• pp.511-518
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• 2018

Initiation of spore germination in filamentous fungi such as Aspergillus nidulans and Botrytis cinerea requires the presence of nutrients. In this study, involvement of sugar sensing machinery was suggested in the germination of A. nidulans spores. Germination did not occur when the spores of A. nidulans were incubated in distilled water, whereas they were successfully germinated in the presence of 5% glucose with a germination rate of over 98% after 6hr incubation. Similar results were obtained when the spores were incubated in the presence of various sugars such as fructose, sucrose, and starch. Interestingly, spore germination was not observed in the presence of D-arabinose, whereas L-arabinose could induce germination as determined by the formation of germ tubes, indicating the presence of sugar sensing machinery that distinguish between the enantiomers of sugars. This inference was further supported by a decrease in germination rate (less than 25%) upon treatment of spores with trypsin. Subsequent MALDI-TOF mass spectrometry analysis of the surface proteins of spores identified ten proteins among which eight were involved in sugar metabolism. Taken together, our results suggest that spore germination in A. nidulans is initiated by the interaction of sugars with sugar binding proteins on the surface of spores.

• Journal of Food Hygiene and Safety
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• v.24 no.3
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• pp.256-261
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• 2009

Bacillus cereus is a gram-positive spore-forming bacterium and produces an emetic or diarrheal syndrome induced by an emetic toxin and an enterotoxin, respectively. In this study, the effect of different types of media, temperature, and time on the sporulation of B. cereus, and thermal resistance of B. cereus spores produced in various temperatures were evaluated. The highest levels of spores were detected when they are produced at $25^{\circ}C$. There were no significant differences in levels of spores produced at $25^{\circ}C$ among culture media and times while levels of spores produced at $43^{\circ}C$ were significantly reduced with the increase of time. However, thermal resistance of B. cereus spores could be affected by incubation temperature. In fact, higher D-values (12.0, 10.1, and 5.9 min for 2,4, and 6 weeks, respectively) of spores produced at $43^{\circ}C$ were observed than did in samples produced at other temperatures (25 and $37^{\circ}C$). D-values of spores were 7.7, 8.2, and 12.0 min when they were produced at 25,37, and $43^{\circ}C$ for 2 weeks, respectively. The sporulation of B. cereus at $25^{\circ}C$ could result in high amounts of spores however the sporulation at $43^{\circ}C$ for 2 weeks could be effective to produce thermal resistant spores.

• The Korean Journal of Pesticide Science
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• v.4 no.4
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• pp.66-71
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• 2000

Effect of temperature on resting spore germination of Plasmodioprora brassicae was indirectly estimated based on examining temporal change of number of inactive resting spores. Resting spore germination was the highest at $28^{\circ}C$ reaching 55.6% and 82.5%, 24hr and 132hr after treatment, respectively. Optimum pH for resting spore germination was pH6, following pH7 and pH8, and the germination was inhibited at pH 4, and pH9. termination of resting spores was stimulated by root extracts of radish, Chinese cabbage and kidney bean, but inhibited by that of lettuce. Number of inactive resting spores was increased as temperature increases and time prolongs after temperature treatment. However, degree of inactivation of resting spores after 1hr at $40{\sim}65^{\circ}C$ was similar with $40{\sim}60%$, but rapidly increased to 91.5% at $70^{\circ}C$. When root galls were submerged in water, density of inactive resting spores was increased rapidly and reached 60.3% 9 days after treatment. Flooding of infested soil resulted in 30% reduction of survived resting spores 5 months later. Among the two registered fungicides, fluazinam was better for inactivation of resting spores than flusulfamide, but both fungicides were inferior to phosphoric acid.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.123-129
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• 2008

To isolate Bacillus cereus presenting at a level of 5 log CFU/g in kochujang, a primary dilution ($10^{-1}$) of kochujang was heated at $85^{\circ}C$ for 5 min. Two isolated strains Voges-Proskauer positive colony (KBC) and a negative colony (KBM) were identified as B. cereus and Bacillus mycoides, respectively, by biochemical test and 16S rDNA sequencing. $D_{100^{\circ}C}$-Values of KBC and KBM strains was 8.37 and 7.08 min, respectively. When spores of KBC strain were inoculated to kochujang at the level of 4-5 log spores/g, the number of spores was no significant difference (p<0.05) for each sample from 1 up to 60 day of aging. When kochujang was inoculated with 4 log spores/g and heated at $85^{\circ}C$ for 15 min, the number of spores was similar to that of unheated kochujang. Therefore, we estimated that B. cereus isolated from kochujang resistant on the heat treatment ($85^{\circ}C$, 15 min) and its heat resistance characteristics could be used to count the number in kochtjang.