• Asian-Australasian Journal of Animal Sciences
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• v.14 no.2
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• pp.163-169
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• 2001

Despite the high potency of the retrovirus vector system in gene transfer, one of the main drawbacks of has been difficulty in preparing highly concentrated virus stock. Numerous efforts to boost the virus titer have ended in unsatisfactory results mainly due to fragile property of retrovirus envelope protein. In this study, to overcome this problem, we constructed our own retrovirus vector system producing vector viruses encapsulated with VSV-G (vesicular stomatitis virus G glycoprotein). Concentration process of the virus stock by ultracentrifuge did not sacrifice the virus infectivity, resulting in more than 108 to 109 CFU (colony forming unit) per ml on most of the target cell lines tested. Application of this high-titer retrovirus vector system was tested on chicken embryos. Injection of virus stock beneath the blastoderms of pre-incubated fertilized eggs resulted in chick embryos expressing E. coli LacZ gene with 100% efficiency. Therefore, our results suggest that it is possible to transfer the foreign gene into chicken embryo using our high-titer retrovirus vector.

• Journal of Microbiology
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• v.34 no.3
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• pp.263-263
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ in CHSE, gradual decrease in $[Ca^{2+}]_i$ in FHM, and no significant change in RTG cells. The degree of $[Ca^{2+}]_i$ increase or decrease was dependent on the amount of infectious virus, and these $[Ca^{2+}]_i$ variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in $[Ca^{2+}]_i$ was not observed. Thus, infectivity of IHNV appears to correlate with changes in $[Ca^{2+}]_i$ in virus-infected cells. These IHNV-induced $[Ca^{2+}]_i$ changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced $Ca^{2+}$ variations were more related with protein synthesis than RNA synthesis. Various $Ca^{2+}$ related drugs were used in search for the mechanisms of the $[Ca^{2+}]_i$, changes following IHNV infection of CHSE cells. Decreasing extracellular $Ca^{2+}$ concentration or blocking $Ca^{2+}$ influx from extracellular media inhibited the IHNV-induced increase in $[Ca^{2+}]_i$, in CHSE cells. Similar results were obtained with intracellular $Ca^{2+}$ blockers. Thus it is suggested that both the extracellular and the intracellular $Ca^{2+}$ sources are important in IHNV-induced $[Ca^{2+}]_i$ increase in CHSE cells.

• Preventive Nutrition and Food Science
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• v.7 no.2
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• pp.168-173
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• 2002

Gastrointestinal infections kill over two million people each year, and pathogen contamination of livestock causes many cases of food poisoning. Two candidate intestinal probiotic strains, L. rhamnosus GR-1 and L. fermentum RC-14 were found to inhibit the growth of Salmonella typhi, Shigella dysenteriae, E. coli O157:H7, Listeria monocytogenes, L. innocua, Enterococcus faecalis, and Bacteroides fragilis. In a series of mouse experiments, L. rhamosus GR-1 and L rhamnosus GG protected against S. typhimurium infection and translocation to the liver and spleen, reduced mortality and induced intestinal phagocytic and immunoglobulin responses. In a second series of experiments, the combination of L. rhamnosus GR-1 and L. fermentum RC-14 was superior to L. rhamnosus GG and placebo in protecting the mice from the lethal effect of salmonella. In summary, the use of combinations of probiotic lactobacilli as dietary supplements or foods could be considered for people at high risk of salmonella intestinal infection. Given the post-infection complications that can arise, such natural methods warrant further exploration especially given the increasing problem of antibiotic resistance and the lack of alternative measures available to many developing countries.

• The Plant Pathology Journal
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• v.18 no.3
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• pp.121-125
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• 2002

This study was conducted to determine the causal virus that naturally infected hollyhock (Althaea rosea) plant showing mild mosaic symptom in 1999. Flexuous virus particles were found in the cytoplasm of plant tissue from infected hollyhock under transmissible electron microscopy. A virus from the genus Potyvirus under the family Potyviridae was isolated and was maintained on Chenopodium quinoa for three passages. Chlorotic local legions were used to inoculate 20 species of indicator plants. The virus infected all the tested cucurbit plants, but failed to infect Nicotiana benthamiana. Based on the host range test and RT-PCR analysis, the potyvirus was identified as a strain of Zucchini yellow mosaic virus-A (ZYMV-A), one of the major pathogens of cucurbits. Infectivity analysis showed that ZYMV-A induced faster systemic symptom than ZYMV-Cu on squash and other cucurbit plants, suggesting that ZYMV-A was a more severe strain. To better characterize ZYMV-A, Western blot assay was carried rout to the coat protein (CP) of the virus using ZYMV-specific antiserum with ZYMV-Cu and other potyviruses. The CP of the virus reacted strongly with the antiserum against ZYMV, and other tested antisera did not react with the CP of ZYMV-A. Results strongly suggest that the potyvirus infecting hollyhock was a novel strain of ZYMV. This is the first report on ZYMV as the causal virus infecting hollyhock in Korea.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.751-759
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• 1999

Plaque characteristics of porcine reproductive and respiratory syndrome (PRRS) virus isolates were examined using MARC-145 line cells. The plaque morphology of PRRS virus isolates was variable in size and heterogenic in population. Upon serial passages of the PRRS virus isolates on MARC-145 tells, heterogeneity was maintained but numbers of the large plaque size virus were increased with certain isolates. A PRRS virus isolate with variable plaque sizes was subcloned into 2 populations : small plaque ($H_S$) and large plaque ($H_L$) viruses. Growth kinetics of the subclones were then determined in MARC-145 cells, and production of the structural polypeptides was analyzed by SDS-PAGE. In a comparison of the growth kinetics, the $H_S$ virus showed higher infectivity titers during the first 48 hours but slower to reach the peak titier than $H_L$ virus did. In a nucleotide sequence comparison, differences of 4 nucleotides in open reading frames 5-6 gene were found between $H_S$ and $H_L$ viruses. Both the $H_S$ and $H_L$ clones produced 5 polypeptide bands with molecular weights of 15, 19, 26, 36 and 42 kD. The 5 bands were detected at 48 hours postinoculation (PI) with antisera to $H_L$ and another large plaque virus ($W_L$) and at 72 hours PI with $H_S$ virus antiserum. The present results demonstrate differences of biologic and molecular characteristics between the two PRRS virus plaque clones.

• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.197-202
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• 1989

This study was performed to observe the susceptibility of dogs and cats as definitive hosts of Paragonimus ilektsuenensis. The metacercariae of this luke were obtained from Sesarma dehaani collected at a focus near the mouth of Sumjin river in November, 1986 and February, 1987. The larvae isolated from the crabs were introduced Per os into 7 albino rats, 2 dogs and 3 cats. The adults were recovered from the experimental animals, End they were morphologically observed and measured. The results were as follows: 1. The recovery rate of adult worms at 42 days after infection was 53.3% from three albino rats, 21.0% from a dog and 12.7% from two cats. Most of the worms were recovered from the worm capsules in the lungs. 2. The size of worms recovered from albino rats, a dog, and cats 42 days after infection aver-aged $6.3{\times}3.2mm,{\;}6.3{\times}3.0mm,{\;}or{\;}6.2{\times}3.5mm$, respectively. There were little differences in the morphology of worms by different experimental animals. 3. The size of eggs from a dog was $88.9{\times}49.3{\mu\textrm{m}}$, and that from cats was $84.3{\times}53.7{\mu\textrm{m}}$ on average. Dogs and cats were good definitive hosts of p. iloktsuenensis. This fact suggests that human infection by this luke may be possible if the metacercariae were ingested.

• Parasites, Hosts and Diseases
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• v.47 no.2
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• pp.131-138
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• 2009

The present study surveyed the prevalence of natural infection of the sheep esphagus muscle with sarcocysts of Sarcocystis ovicanis and examined induction of protective immunity using UV-attenuated sporocysts. The overall prevalence of natural infection of the sheep was 95%. Infectivity of the collected sarcocysts was confirmed by shedding of sporulated oocysts after feeding infected esophageal tissues to dogs. To induce protective immunity, lambs were immunized 3 times (once a week) with $1.5{\times}10^4$ sporocysts exposed to UV-light for 30min (UV-30 group) or 60 (UV-60 group) min and then challenged with $1.5{\times}10^4$ normal sporocysts at the 3rd week post the 1st vaccination. These lambs showed high survival and less clinical signs of sarcocystosis than normal infected lambs. The attenuated sporocysts produced abnormal cysts; small in size and detached from the muscle fiber. These abnormalities were more obvious in UV-60 group than UV-30 group. Also, the $IFN-{\gamma}$ level and lymphocyte percentage were increased while the total leukocyte count was decreased in the UV-60 group compared with other groups. The high level of $IFN-{\gamma}$ may be an evidence for the induction of Th1 responses which may have protective effect against a challenge infection.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.171-180
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• 2001

Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10$^{8}$ CFU/ $m\ell$ on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100$\times$) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.33 no.4
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• pp.380-385
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• 1988

The present study was conducted to evaluate agronomic characters of the soybean germplasm which was collected from the sites covering whole areas of southern Korean peninsula. A total of 2,748 Korean native soybeans was grouped for fruiting period and in this group frequency distribution of the important agronomic characters and relationships among the characters were determined. The results obtained were summarized as follows; The soybean germplasm was comprised of SeoulㆍKyonggi-do 7.5%, Kangwon-do 13.3%, Chungchongbuk-6.6%. Chungchongnam-do 12.3%, Chollabuk-do 10.7%, Chollanam-do 12.6%, Kyongsangbuk-do 20.8%, Kyongsangnam-do 15.3% and Cheju-do 0.6% in number of collections. Frequency distribution of the germplasm for fruiting period was 0.14% in Group I (below 55 days). 1.67% in Group II (56-60 days), 10.58% in (Group III(61-65 days), 23.18% in Group IV(66-70 days), 25.91% in Group V(71-75 days) 19.79% in Group Ⅵ(76-80 days), 15.1% in Group Ⅶ(81-85 days) and 3.6% in Group Ⅷ(over 85 days). The lines over 70 days in fruiting period were as much as 64.4% of germplasm. The fruiting period varied largely from 45 days to 91 days. Fruiting period was positively correlated to seed weight, days to maturity, days to flowering, plant height and number of branches per plant, respectively. While. it was negatively correlated to number of pods, lodging and virus infectivity, repectively, and in particular negative correlation was shown significantly between yield and fruiting period. Average yield was very high as much a 1.389kg/ha in fruiting period Group I, and decreased with increment of fruiting period, 1.400kg/ha in Group II. 1.384kg/ha in Group III. 1,299kg/ha in Group IV, 1,197kg/ha in Group V, 1,117kg/ha in Group Ⅵ, 967kg/ha in Group Ⅶ and 832kg/ha in Group Ⅷ.

• Journal of the Korean Home Economics Association
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• v.30 no.3
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• pp.91-99
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• 1992

Cell structure is based on proteins. Since the antibody is proteous substance, the continous low protein feeding decreases the resistance of host against pathogenic agents. The present study was designed to investigate the infectivity of protozoa to rats which were fed with variously prescribed diets. Experimental group was divided into 4 groups according to the level of casein in the diet, group I: casein 0%, group II: casein 5%, group III: casein 15%, group IV: casein 30%. Each animal was fed for 5 weeks followed by inoculation of protozoa in cecum and sacrified each 1 week later of the infection. Each diet group, non infected with protozoa was recognized as the control. Result are summerized as follows : 1. All the rats of group I died in 2∼4 weeks and 2 of 12 rats in group II were also died in the period. 2. The growth rate and FER were high in group III and IV compared with group II. Therefore low protein feeding decrease growth and feed efficaly ratio(FER). 3. The pH of caecal contents between the infected group and control showed no difference, but the values of group III and IV were higher than the group II. Low pH of the caecal contents provides a suitable condition for determining their susceptibility to Entameoeba histolytical trophozoite. 4. Amounts of serum total protein in group II, III and IV showed no significant difference with the control and infected group, but amounts in group III and IV were higher than the group II. Therefore, continuous low protein feeding decrease serum total protein. 5. Albumin, ${\alpha}$1, gloulin, ${\alpha}$2 globulin, ${\beta}$ globulin, ${\gamma}$ gloulin of group III and IV were all high to compare than the group II. Albumins of group III and IV of control was higher than infected group, but there was no difference in ${\gamma}$ globulin between the infected and control group.