The pathogenicity of free-living amoeba, Waegleria fcwleri, is influenced according to the strain, cultural condition and host (Culbertson et at., 1968; Carter, 1970; Wong et at., 1975), Phillips (1973) demonstrated that Entamoeba histolytica became avirulent after more than 2 year maintenance in axonic culture in vitro. This study was carried out to compare the difference in pathogenicity between two strains of N. fowleri, one of a prolonged maintenance in arsenic medium and the other one obtained by serial brain passage in mice. The 0 strain was that N. fowleri had cultivated axenically more than 7 years in CGVS medium. The 2-1 strain was obtained from the brain of mouse inoculated intranasally with a strain, which was from the mouse brain infected with 0 strain, and cultured for 15 weeks until the beginning of this experiment. White male mice weighing 18-22 g were used. Mice were anesthetized by an intraperitoneal injection of about 1 mg secobarbital, and inoculated intranasally with $10{\times}$10^4 live N. fowleri trophoBoites in a $5{\;}{\mu}l$ cell suspension. Sluggish behaviour, nervousness, rotation and leg paralysis were developed earlier and more frequently in the 2-1 experimental group than the control 0 group. Pathological changes such as inflammatory and necrotic lesion were observed in the olfactory and anterior portion of brain, and these changes were more extensive in the 2-1 group. The edematous and inflammatory changes in lung were demonstrated in mice died after 13th day post-inoculation. The experimental mice of 2-1 group began to die suddenly from 7th day post-inoculation, and the survival time in 2-1 group mice was shorter than 0 group mice. The typical primary amoebic meningoencephalitis was developed in the mice inoculated intranasally with N. fowleri. The prolonged maintenance of N. fowleri amoebae in axonic CGVS medium was observed to have lost their original pathogenicity for mice, but their pathogenicity was restored by serial brain passage in mice.
Orchids have been propagated vegetatively for a long time without adequate control measures against virus diseases in Korea. As a result, it is presumed that most of the orchid varieties in Korea may have been degenerated. Nevertheless there has been little work on the virus diseases of orchids in Korea. Therefore studies were initiated to isolate an4 characterize the orchid viruses occurring in Korea. The results obtained are summerized as follows. 1. Symptoms of virus diseases on orchid varieties can be grouped 1) mosaic, 2) necrotic streak with mosaic, 3) ring necrosis, 4) chlorotkc ring and 5) necrotic spot. 2. A total of 102 orchid plants representing 4 genera were investigated on the occurrence of Cymbidium mosaic virus and tobacco mosaic virus by serological agar-gel double diffusion test. The test revealed that approximately $45\%$ of the orchids were infected with Cymbidium mosaic virus. None of the plants were found to be infected with tobacco mosaic virus. 3. Local lesions appeared on the inoculated leaves of Chenopodium amaranticolor Cassia occidentalis and Datura stramonium 7-12 days after mechanical inoculation with Cymbidium mosaic virus. 4. Physical properties of the Cymbidium mosaic virus determined by inoculation on Chenopodium amaranticolor were as follows: Thermal inactivation Point; $75-80^{\circ}C$, dilution end Point; $10^{-5}-10^{-6}\%$ aging in vitro; 8 days. 5. Three different buffers at pH 7.0 and pH 9.0 were compared for the efficiency of agar-gel double diffusion test with Cymbidium mosaic virus. Phosphate, imidazol and tris buffer at pH 7.0 gave equally satisfactory results. 6. Electron microscopic examination of the Cymbidium mosaic virus revealed rod shaped particles measuring 460-580mu.
To develop an effective biopesticide to control pepper anthracnose disease, an isolate which showed strong inhibitory effect on the mycelial growth and conidial germination of Colletotrichum acutatum was selected among the antagonistic bacterial isolates collected from pepper grown soil. The bacterial isolate was identified as Bacillus sp. KBC1004 using 16S rRNA sequence analysis. The liquid culture of KBC1004 was freeze-dried and formulated as a wettable powder(WP). The wettable powder form of KBC1004 required at least 24 hours to activate and to inhibit the conidial germination of C. acutatum. In vitro bioassay using the detached green pepper fruits, biocontrol activity of the WP was not recognizable in simultaneous inoculation, but significant disease suppression was observed pre-treatment (24 hr) of the WP before pathogen inoculation. In field experiment, 4 times foliar applications of the 1/500 diluted wettable powder from the end of June showed great control efficacy similar to that of the chemical fungicide application. These results suggest that the formulated WP product could be an alternative mean to control of pepper anthracnose disease in environmentally friendly farming practices.
The prevalence of the infectious flacherie virus (FV) disease causes a severe damage to cocoon yield and various methods to control the disease have been studied. In this regard, guanidine hydrochloride (GH), one of the guanidine derivatives known as the most inhibitory agent against the replication of picorna virus, was applied to silkworms per os with mulberry leaves and the results were as follows. 1. The application of GH below 0.01% of the chemical concentration did not give any damage to silkworm larvae. 2. The transmission of the virus disease by introducing the FV infected larvae to the healthy larvae group was proportioned to the number of infected larvae. When l% of infected larvae was introduced to the rearing tray of healthy larvae, the pupation rate was 70.7%(79) and it was 38.4% (43) to 5% of infected larvae introduced, while the control of non-mixed with infected larvae gave 89.2% (100) of pupation rate. The cocoon yield from 10,000 larvae also showed the same tendency as the pupation rate. 3. The inhibitory effect of GH against the replication of FV showed ten times in treatment of 0,01% of the chemical agent compared to the non-treatment. 4. The successive application of GH after virus inoculation to silkworm larvae led to the most effective on the inhibition of the virus replication. 5. The immediate application of GH after the virus inoculation also gave the best effect on the inhibition of the virus replication in silkworm larvae. 6. The effect of GH on the inactivation of FV in vitro was not observed.
Ka, Kang-Hyeon;Park, Hyun;Hur, Tae-Chul;Bak, Won-Chull
The Korean Journal of Mycology
/
v.36
no.2
/
pp.148-152
/
2008
We inoculated hypal suspension of Tricholoma matsutake and T. magnivelare were examined on Pinus densiflora seedlings grown in a granite soil substrate with 1/2 PDMP (12 g/l potato dextrose broth, 1.5 g/l malt extract, and 0.5 g/l peptone) medium. Four months after inoculation, the pine seedlings were examined for infection rate, matsutake aroma, and Hartig-net formation. The roots of pine seedling formed ectomycorrhizal roots in the 9 isolates from 12 isolates of T. matsutake and T. magnivelare. However, the seedlings showed different ectomycorrhizae forming rates among the 9 isolates. While matsutake aroma was confirmed from the ectomycorrhizal seedlings, the pine seedling contaminated by bacteria or fungi did not form matsutake ectomycorrhizae with sickening smell. Thus, the aroma was chosen as a good way for the verification of mycorrhizal infection. At the early stage, the mycorrhizal roots showed unramified and branched types without root hair. They also showed thin mantle layers, Hartig-nets, and turned into black color at later stage. Among the examined strains, that of Yecheon isolated in 1995 showed the best infection rate, which indicated that we need to pay attention to the selection of isolates for better result.
Cha, Seung-Bin;Rayamajhi, Nabin;Lee, Won-Jung;Shin, Min-Kyoung;Roh, Yu-Mi;Jung, Myung-Hwan;Myoung, Kil-Sun;Ahn, Young-Tae;Huh, Chul-Sung;Yoo, Han Sang
Korean Journal of Veterinary Research
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v.50
no.3
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pp.213-220
/
2010
Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.
Journal of The Korean Society of Grassland and Forage Science
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v.36
no.1
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pp.7-14
/
2016
The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed.
Kim, Su Jun;Kim, Eun Su;Kim, Seung Heui;Yun, Hae Keun
Korean Journal of Agricultural Science
/
v.45
no.3
/
pp.365-378
/
2018
Anthracnose caused by Colletotrichum gloeosporioides, which is one of the major diseases of red dates, causes severe damages in jujube (Zizyphus jujuba Miller) production in Korea. This study was done to evaluate the inhibition of anthracnose occurrence and pathogen growth by the treatment of environment-friendly materials such as a Bordeaux mixture and loess-sulfur mixture and by defense-response signaling in jujube. The in vitro test of the environment-friendly materials and signaling molecules that were routinely applied did not exhibit any antifungal activities against the pathogen for jujube anthracnose. The Bordeaux mixture and loess-sulfur mixture at a two-fold concentration showed inhibition zones that were 16.0 and 20.3 mm in diameter, respectively. In the pathogen inoculation test with detached jujube tree leaves, while treatment with the environment-friendly materials diluted by half showed no inhibition of lesion development, they did show inhibition of lesion development when they were routinely applied to the leaves. In detached jujube fruits inoculated with the pathogen, better suppressive effects by the treatment of the environment-friendly materials were seen in the fruits at a young stage rather than in the ripening stage. The in vivo test with jujube trees in pots showed that the treatment of salicylic acid (1 mM) resulted in the best suppressive effects against lesion development. The results suggest that it is possible to manage the incidence of anthracnose by the treatment of environment-friendly materials such as the Bordeaux and loess-sulfur mixtures and signaling chemicals such as ethephon, hydrogen peroxide, methyl jasmonate, and salicylic acid in jujube trees and fruits. Consequently, these findings suggest that environment-friendly materials and defense response signaling molecules could be used as suitable candidates for sustainable agrochemicals to manage anthracnose in jujube production.
A rapid radicle assay for prescreening antagonistic bacteria to Phytophthora capsic4 causal agent of Phytophthora blight of pepper was developed. Sixty-four bacterial strains with in vitro antifungal activity selected out of 1,400 strains isolated from soils of Ansung, Chunan, Koyang, and Paju, Korea in 1998 were used for development of the bioassay. Uniformly germinated pepper seeds dipped in bacterial cells for 3 hours were placed near the edges of growing mycelia of P. capsici on water agar containing 0.02% glucose. Five-week-old pepper plants(cv. Nockwang) were inoculated to compare with results of the radicle assay developed in this study. For plant inoculation, pepper seeds were sown in potting mixtures incorporated with the bacterial strains, then transplanted into steam-sterilized soils 3 weeks later. Plants were hole-inoculated with zoospores of P. capsici 2 weeks after transplanting. Disease incidence and severity were determined in radicle and plant assessments, respectively. In radicle assay, six strains, GK-B15, GK-B25, OA-B26, OA-B36, PK-B09, and VK-B14 consistently showed the significant(P=0.05) disease reduction against radicle infection by the fungus, four of which also did in plant assessments. Strains OA-B36 and GK-B15 consistently reduced the fungal infection in both the radicle assay and the plant assessment. Therefore, prescreening strains using the radicle assay developed in this study followed by plant assay could reduce time and labor, and improved the possibility of selecting antagonistic bacteria for control of Phytophthora blight of peppers.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.283-283
/
2017
The objective of the present study is to analyze the lactic acid bacteria (LAB) effects on rye silage fermentation at different stages. Different stages (Booting, Heading, Flowering, and Late flowering stage) of rye were collected from the National livestock farm, National Institute of Animal Science, South Korea. Rye sample was inculcated with lactic acid bacteria and incubated at the anaerobic condition for three months. The nutrient profile such as crude protein (CP), Acid detergent fibre, Neutral detergent fibre and total digestibility nutrients were increased in both control and LAB inculcated samples at all the stages of rye forage. The pH of rye silage was reduced at both stages by LAB inoculation as compared with control. The lactate content was increased in all stages of rye sample by LAB. The acetate concentration and butyrate was reduced in LAB inoculated rye sample. However, acetate concentration was slightly high in LAB inculcated rye at heading and late flowering stage. The LAB population was greater in LAB inoculated rye sample as compared with control sample. However, the massive population was noted in booting stage of rye than the other stages. It indicates the inoculated LAB is the main reason for increasing fermentation quality in the sample through pH reduction by lactate production. Overall results suggest that the isolated lactic acid bacterium is the potent strain that could be suitable for rye forage fermentation at different stages.
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