In this study, we investigated the effect of fermentation conditions on the amylolytic and proteolytic activities of Aspergillus luchuensis strain 74-5 and Aspergillus oryzae strain 75-2, which are used in the preparation of the starter culture, for Takju (Korean traditional rice wine). The starter culture was optimized using different conditions, such as inoculum size, inoculation temperature, and incubation time. The enzyme activities under each condition were measured. In the A. luchuensis strain 74-5 starter culture, the ${\alpha}-amylase$ and glucoamylase activities increased, however the activity of acidic protease decreased as the diluent to starter culture ratio increased. In the A. oryzae 75-2 starter culture, all enzyme activities were maintained at a higher level even at 5% inoculation ratio. Higher enzyme activities were observed in the middle range of inoculation temperature (35, $40^{\circ}C$), than in the lower range (20, $30^{\circ}C$). Enzyme activity in the starter culture varied with incubation time, however it was the highest at 144 and 120 hr, respectively, for A. luchuensis strain 74-5 and A. oryzae strain 75-2. The spore count of the starter culture was approximately $2{\times}10^7$ during fermentation, out of which contamination by aerobic bacteria was about $3{\times}10^3$. The results suggested that the starter culture of each strain could be used as an inoculum for fermentation. However, we needs to conduct further research for the selection of suitable diluting agents as well as drying methods to reduce the contamination by aerobic bacteria, while retaining the enzyme activity.
Park, Whan-Jun;Jwa, Mi-Kyung;Hyun, Sun-Hee;Lim, Sang-Bin;Song, Dae-Jin
Journal of the Korean Society of Food Science and Nutrition
/
v.35
no.10
/
pp.1449-1455
/
2006
Raw oysters were treated at $10^{\circ}C$ and $22^{\circ}C/350$ Mpa/15 min, and microbial counts and quality were measured during storage of 14 days at $10^{\circ}C$. Total viable cell count (TVCC) in untreated oyster increased greatly during storage from starting inoculum of $1.6\times10^2\;CFU/mL$, and reached to $5.6\times10^2\;CFU/mL$ after 4 days of storage. TVCC of the pressure-treated was about $10^1\;CFU/mL$ right after high hydrostatic pressure treatment and increased slowly during storage, and about $10^3\;CFU/mL$ even after 7 days of storage. Lactic acid bacteria count (LABC) in the untreated was increased greatly during storage from starting inoculum of $3.3\times10^3\;CFU/mL$ at 3 days of storage and $7.2\times10^4\;CFU/mL$ after 4 days of storage. LABC in the pressure-treated was detected only after 5 days of storage, and about $10^2\;CFU/mL$ after 8 days of storage. The pH of the untreated was 6.19 and decreased gradually during storage, and 5.83 after 4 days of storage. The pH of the pressure-treated showed little change during storage, and 6.07, 6.03 and 5.82 after storage of 4, 8 and 14 days, respectively. Volatile basic nitrogen (VBN) in the untreated was 16.8 mg%, and maintained almost constant until 1 day of storage, and then increased suddenly, and 30.1 mg% after 4 days of storage. VBN of the pressure-treated stayed unchanged during storage, and about 20 and 23 mg% even after 4 and 8 days of storage, respectively. Hunter $L^*,\;a^*\;and\;b^*$ values were increased until 2 days of storage and then showed no change during storage. Demerit score was the lowest in the thawed raw oyster, and then in the increasing order of the pressure-treated (4 day and 8 day storage) and the untreated (4 day storage).
To reveal the immunogenicity of ${\gamma}-irradiated$ E tenella and its progeny, a series of experiments on the effects of Cobalt-to ${\gamma}-irradiation$ was performed. The SPF chickens inoculated with diffenrt doses of inoculum were challenged with $1{\times}10^5$ oocysts of virulent E tenella. The levels of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and of inoculum with $1{\times}10^4$ oocysts were recognized as proper as immunogen by comparison of survival rates, body weight gains, blood in feces and lesion scores in the chickens. In these trials of challenge with virulent E tenella after inoculation with $1{\times}10^4$ oocysts of the ${\gamma}-irradiated$ E tenella and its progeny, the survival rates of the chickens challenged with the virulent E tenella after immunization with the 1st and the 3rd progeny groups of ${\gamma}-irradiated$ E tenella oocysts were higher(l00%) than that(87.0%) of the challenged control group. The signs of blood in feces and the lesion scores were seen markedly lower with the ourput of the smaller number of oocysts, i.e. OPG 103,900 and 25,800 in the groups of the 1st and the 3rd progeny, respectively, than those(OPG 1,658,900) of the challenged control group. The body weight gains of the 1st and the 3rd progeny groups, the 1st week and the 2nd week after challenge, were higher (2.6g and 155.4g, 11.6g and 168.9g respectively) than those(-85.8g and 63.6g, respectively) of the challenged control group, and the feed conversion ratios(FCR 3.28 and 2.96) of the 1st and the 3rd progeny groups were lower than that(FCR 5.60) of the groups challenged control group. The anticoccidial indices(70.5 and 93.9) of the groups challenged with the virulent oocysts of E tenella after immunization with the 1st and the 3rd progeny of the ${\gamma}-irradiated$ E tenella were significantly higher than that (ACI -81.9) of the challenged control group. It was thought that the immunogenicity of ${\gamma}-irradiated$ E tenella would be increase according to increase the number of generation passaged in chicken. That might be because of increasing the pathogenicity of ${\gamma}-irradiated$ E tenella according to increase the number of generation passaged in chicken.
A series of experiments on the effects of ${\gamma}-irradiation$ was performed to reveal the pathogenicity of ${\gamma}-irradiated$ oocysts of E tenella from Cobalt-60 and its progeny. The SPF chickens were inoculated with differnt doses of radiation and inoculum. The level of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and the level of inoculum with $1{\times}10^4$ oocysts were recognized more pathogenic than those of the other groups by comparison of body weight gains, blood in feces and lesion scores. The signs of blood in feces, lesion score and the number of excreted oocysts in the feces were revealed as the lowest in the group of the ${\gamma}-irradiated$ oocysts, the average in the group of the 1st and the 3rd progeny, and the highest in the group of non-irradiated oocysts of E $tenell\grave{a}$. The body weight gain of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was higher than those of the non-irradiated, the 1st and 3rd progeny groups. The body weight gain of the groups immunized with the 1st and the 3rd progeny of E tenella were higher than that of the non-irradiated group. The feed conversion ration of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was lower than those of the non-irradiated, the 1st and the 3rd progeny groups. The feed conversion ratios of the group immunized with the 1st and 3rd progeny of ${\gamma}-irradiated$ oocysts were lower than that of the group infected with non-irradiated E tenella. The anticoccidial index(ACI 190.6) in the chickens immunized with the ${\gamma}-irradiated$ oocysts of E tenella and those(ACI 142.8 and 107.4) of the 1st and the 3rd progeny groups were higher than that (ACT 87.4) of the group infected with non-irradiated E tenella. It was thought that the pathogenicity of ${\gamma}-irradiated$ E tenella would be recovered according to increase the number of generation passaged in chicken.
Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
Horticultural Science & Technology
/
v.33
no.1
/
pp.70-82
/
2015
This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.
Park, Whan-Jun;Jwa, Mi-Kyung;Hyun, Sun-Hee;Lim, Sang-Bin;Song, Dae-Jin
Journal of the Korean Society of Food Science and Nutrition
/
v.35
no.7
/
pp.935-939
/
2006
Raw oyster (Crassostrea gigas) was inoculated with Vibrio parahaemolyticus and Escherichia coli, treated with high hydrostatic pressure and evaluated for microbial counts. Cell death of V. parahaemolyticus (Vp) increased with the increase of applied pressure. Vp starting inoculum of $3.8{\times}10^5\;CFU/mL$ was totally eliminated after exposure to 200 MPa for 10 min at $22^{\circ}C$ Viable cell of Vp decreased with the increase in treatment time and dropped below the detection limit with treament of 25 min at $22^{\circ}C/150\;MPa$. The number of Vp by treatment of $0^{\circ}C$ and $10^{\circ}C$ for 20 and 25 mon at 100 MPa, respectively. For E. coli, there was an initial lag up to 250 MPa gollowed by a rapid decline. Treatment at 325 MPa/$22^{\circ}C$ for 15 min caused 5-log reduction, while that at 375 MPa resulted in total reduction of starting inoculum of $4.0{\times}10^7\;CFU/mL$. Lower treatment temperature showed higher killing effect of E. coli at the same treatment pressure and time. Viable cell of E. coli decreased with the increase in treatment time, and 4-log reduction was achieved with treatment of 5 min at $10^{\circ}C$/350 MPa and then total reduction was achieved after treatment of 15 mon. Higher pressure, lower temperature and longer time were more effective in sterilizing V. parahaemolyticus and E. coli.
This study was carried out to investigate the effect of ${NO_3}^-$ and ${NH_4}^+$ on the adventitious root growth and eleuthroside synthesis of Eleutherococcus koreanum in 5 L-bioreactor culture. The change in the medium components was also measured during culture. The fresh weignt of adventitious root reached to the highest level of 30.8 g FW/L in the presence of both 50 mM ${NO_3}^-$ and 10 mM $NH_4^+$, representing 3.6-fold increase compared to the 60 mM ${NH_4}^+$ alone. However, as the increase of the portion of ${NH_4}^+$, the root growth was decreased. However, the maximum eleutheroside B, E and E1 contents were $57.3{\mu}g/g$ DW, $188.4{\mu}g/g$ DW and $47.3{\mu}g/g$ DW, with 30 mM, 60 mM and 15 mM total nitrogen source, respectively. Fresh weight of adventitious root increased up to 6.8-fold of inoculum size within 9 weeks. The amounts of ${NH_4}^+$, $K^+$, ${NO_3}^-$ and ${PO_4}^-$ were decreased during culture periods. Based on these results, we suggest that various further studies are required to increase the biomass and the useful secondary metabilites.
The objective of the experiment was to determine the optimum cultural [moisture levels (55, 60 and 70%), days of fermentation (7, 14 and 21), temperature (25 and $35^{\circ}C$) of incubation)] and nutritional parameters (urea addition (0 and 2%) and variable levels of single super phosphate (0.25 and 0.50% SSP)) for bio-processing of the mustard (Brassica campestris) straw (MS) under solid-state fermentation (SSF) system. The performance of SSF was assessed in terms of favorable changes in cell wall constituents, protein content and in vitro DM digestibility of the MS. Sorghum based inoculum (seed culture) of Ganoderma lucidum to treat the MS was prepared. The 50 g DM of MS taken in autoclavable polypropylene bags was mixed with a pre-calculated amount of water and the particular nutrient in the straw to attained the desired levels of water and nutrient concentration in the substrate. A significant progressive increase in biodegradation of DM (p<0.001), NDF (p<0.01) and ADF (p<0.05) was observed with increasing levels of moisture. Among the cell wall constituents the loss of ADF fraction was greatest compared to that of NDF. The loss of DM increased progressively as the fermentation proceeded and maximum DM losses occurred at 28 days after incubation. The protein content of the treated MS samples increased linearly up to the day $21^{th}$ of the incubation and thereafter declined at day $28^{th}$, whereas the improvement in in vitro DM digestibility were apparent only up to the day $14^{th}$ of the incubation under SSF and there after it declined. The acid detergent lignin (ADL) degradation was slower during the first 7 days of SSF and thereafter increased progressively and maximum ADL losses were observed at the day $28^{th}$ of the SSF. The biodegradation of DM and ADL was not affected by the variation in incubation temperature. Addition of urea was found to have inhibitory effect on fungal growth. The effect of both the levels (0.25 and 0.50) of SSP addition in the substrate, on DM, NDF, ADF, cellulose and ADL biodegradation was similar. Similarly, the protein content and the in vitro DM digestibility remain unaffected affected due to variable levels of the SSP inclusion in the substrate. From the results it may be concluded that the incubation of MS with 60 percent moisture for 21 days at $35^{\circ}C$ with 0.25 percent SSP was most suitable for MS treatment with Ganoderma lucidum. Maximum delignification, enrichment in the protein content and improvement in in vitro DM digestibility were achieved by adopting this protocol of bioprocessing of MS.
Perchlorate ($ClO_4^-$) is a contaminant found in surface water and soil/ground water. Microbial removal of perchlorate is the method of choice since microorganisms can reduce perchlorate into harmless end-products. Such microorganisms require an electron donor to reduce perchlorate. Conventional perchlorate-removal techniques employ heterotrophic perchlorate-reducing bacteria that use organic compounds as electron donors to reduce perchlorate. Since continuous removal of perchlorate requires a continuous supply of organic compounds, heterotrophic perchlorate removal is an expensive process. Feasibility of autotrophic perchlorate-removal using elemental sulfur granules and activated sludge was examined in this study. Granular sulfur is relatively inexpensive and activated sludge is easily available from wastewater treatment plants. Batch tests showed that activated sludge microorganisms could successfully degrade perchlorate in the presence of granular sulfur as an electron donor. Perchlorate biodegradation was confirmed by molar yield of $Cl^-$ as the perchlorate was degraded. Scanning electron microscope revealed that rod-shaped microorganisms on the surface of sulfur particles were used for the autotrophic perchlorate-removal, suggesting that sulfur particles could serve as supporting media for the formation of biofilm as well. DGGE analyses revealed that microbial profile of the inoculum (activated sludge) was different from that of the biofilm sample obtained from enrichment culture that used sulfur particles for $ClO_4^-$-degradation.
Bacterial diseases of soybean has been recognized as a limiting factor of soybean production in Korea as it was estimated to cause around 10 percent of yield losses annually. The purpose of the study is to obtain information on the diseases through proving the kinds of pathogens and epidemiology, The wire brush method and multineedle appeared to be the best way of inoculation under all circumstances. Wire brush method, especially, was effective in shortening the incubation period and manifesting the lesion development by introducing more inoculum per unit of area. In case of spray inoculation it was necessary to apply a small amount(1 : 1,000) of wetting agent, twin-20, otherwise it was unabled to produce the diseases under field conditions. Two kinds of bacterial diseases caused by Pseudomonas glycinea and Xanthomonas phaseoli var. sljense were found from surveyed areas in Kore. Wild fire disease on soybean caused by Pseudomonas tabaci had not detectable during the experiment although there were several reports on the disease from other countries. when the pathogens were introduced into sterile soil, bacterial leaf blight pathogen could exsisted until 30 days while bacterial pustule pathogen survived only 4 days under the natural conditions of later June. Both bacteria, however, could produce the disease after more than 10 months period of storage in refrigerator when they were exsisted in infected plant tissues. In warehouse, non-temperature controlled, the bacteria lose their infectability within 6 months period from October to April even though they exsisted in infected tissues. Surface infested seeds with the pathogens could not produced the diseases on seedling stages of soybean plants when the seeds were planted in sterile soil after inoculation by dipping the seeds into bacterial suspensions, although germination was depressed by the pathogens when the seeds were planted on agar media.
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