• Food Science of Animal Resources
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• v.29 no.2
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• pp.151-156
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• 2009

The prevalence of food allergies was investigated using questionnaires with 300 subjects whose ages ranged from 19 to 24 years old and the causative food allergens was analyzed using immunological analysis with serum of the subjects who answered that they have/had food allergy. The questionnaire showed that 11.33% of subjects have/had experience of food hypersensitivity, where the main causative foods were fish, beef, chicken, milk, egg, and pork in order. The meat allergy shared 4.65% (2.33% for beef, 1.66% for chicken, 0.66% for pork) in the prevalence of food allergies. The causative beef allergens were investigated with the serum of 6 subjects who have had beef allergy. Western blots were carried out with the serum of P6 subject who showed a positive reaction to beef extract in ELISA. The two specific bands were detected in beef extract on the PVDF membrane, and no band was detected in extracts of pork and chicken. A calculation of the distance of migration by SDS-PAGE enabled the molecular masses of the two bands to be estimated as 67kDa and 31kDa, respectively. The 67kDa was revealed as bovine serum albumin (BSA) which is one of the important beef allergens as reported previously though an analysis of the N-terminal amino acid sequence. However we could not identify the sequence of 31kDa, probably because they comprised several subunits and were modified proteins such as glycoprotein that were unlikely to be easily degraded by the Edman method. The 31kDa band were dyed with the PAS (periodic acid-schiff reagent), suggesting that it might be a glycoprotein. These results suggested that the 31kDa might be considered as a novel potential beef allergen which is not reported previously, although further studies are needed.

• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.486-492
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• 2007

Seminal plasma(SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs(1-4 years old) of various breeds were pooled, centrifuged at $300{\times}g$ for 10 min at $25^{\circ}C$, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris(EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to $4^{\circ}C$ for 2h or held at $25^{\circ}C$ as controls. Sperm concentration was adjusted to $2{\times}10^8/ml$. [Exp II] Sperm samples, each of which contained $1{\times}10^8/ml$, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT+0.6M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at $25^{\circ}C/min$ of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at $38^{\circ}C$ for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at $200{\times}$ magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6M glycerol containing SP exhibited the higher progressive motility before being frozen(P<0.05). However, frozen-thawed spermatozoa that had suspended in EYT+0.6M glycerol containing SP showed the similar or lower viability(P<0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant(CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.67-83
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• 1983

The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.

• Parasites, Hosts and Diseases
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• v.27 no.4
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• pp.231-248
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• 1989

The growth and development of the metacercariae of F. seoulensis cultivated in vitro or on the chick chorioallantois were assessed by comparison with the optimum process of maturation in albino rats and new born chickens. The process of maturation was divided for convenience into six stages: Stage 1 ; cell multiplication, Stage 2; body shaping, Stage 3; separation of genital anlagen, Stage :1 organogeny, Stage 5; gametogony, and Stage 6: oviposition. In Hank's and Tyrode's .solutions, the metacercariae were alive up to 200 days or more at $4^{\circ}C$ without any development. The in vivo maturation process in rats or chicks was as follows: stage 1 from 6 hours; stage 2 from 24 hours; stage 3 from 48 to 72 hours; stage 4 from 3 to 4 days; stage 5 from 4 to 5 days; and stage 6 from 5 to 8 days. Despite unsuccessful infection of the metacercariae to 12 day old chicks, fully mature worms of stage 5 or 6 were recovered from new born chicks (1 to 2 days old), The metacercariae of F. seoulensis grown in vitro were up to stage 3 and no further maturation was observed. Of various media employed, the medium NCTC 109 (Gibco) or NCTC 135(Gibco) supplemented with 20% egg yolk or 20% whole egg macerate or 0.5% yeast was basically required for the earlier development of the fluke. It took 16.1 days(in average) to reach the stage 3 after cultivation. The metacercariae cultivated on the chorioallantoic membranes of 6∼13 day old chick embryo at 37∼38℃ showed their full development up to stage 5 or 6. However, the worms were in general remarkably retarded, compared with those grown in rats or chickens. In the experiments of worm transplant, although the transfer was failed from in vitro culture to in vivo of rats(Per os), the transplants from in vitro culture to the chorioallantois and from the choriollantois to in vivo of rat host were successful with or without development of the transferred worms. In the present study, it was observed that the metacercariae of F, seoulensis can be maintained in vitro media with poor development as well as fully matured in 1 to 2 day-old chicks or on the chorioallantois at a very low rate.

• Korean Journal of Veterinary Research
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• v.15 no.1
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• pp.57-67
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• 1975

The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.155-159
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• 2013

This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of $2{\times}10^8/ml$ by doubling in every 30 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm ($93.27{\pm}1.62%$) than frozen-thawed sperm ($73.34{\pm}3.27%$). However, there were no significant differences between fresh and frozen-thawed dead cell rate ($7.35{\pm}2.63$ vs, $13.71{\pm}2.85$). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different ($72.86{\pm}2.83$ vs, $81.47{\pm}2.48$), similarly, the dead cell rates was similar ($18.41{\pm}3.42%$ and $17.26{\pm}4.25$). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

• Korean Journal of Ichthyology
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• v.11 no.1
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• pp.94-101
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• 1999

This study was conducted to observe the early life history of sea bass, Lateolabrax japonicus. The fertilized eggs were spherical in shape and turned out to be separative and floated. Their membrane and yolk having 1~5 oil globule were transparent. Fertilized eggs were measured to be 1.33~1.46 mm in diameter. Hatching of eggs were started at 74 hrs 15 mins, 54 hrs 55 mins, 50 hrs 45 mins, after fertilization in water temperature $16.0^{\circ}C$, $18.0^{\circ}C$, $20.0^{\circ}C$ respectively. The newly hatching larvae were 3.79~3.97 mm in total length with 35~37 myomeres, and mouth and anus were closed. Melanophores were distributed on the up side of head, upper jaw and margins of the body. The 5 days after hatching larvae measured 4.78~5.24 mm in total length, yolk were completely absorbed, and transformed to postlarval stage. In this time, mouth of larvae was opened, and also melanophores were presented on the lower jaw. Head of larvae grew remarkably. The 21~22 days after hatching, total length of the larvae was 7.15~8.12 mm, the caudal fin rays began to differentiation. In 31 days after hatching, the larva were 8.46~9.16 mm in total length, and tip of the caudal notochord flexed $45^{\circ}$. The larvae reached to the juvenile stage with all the fins were developed at 61 days after hatching and attained 16.28~17.31 mm in total length.

• Korean Journal of Ichthyology
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• v.2 no.2
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• pp.169-181
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• 1990

The development of eggs and larvae, and minute scale-like tubercles on the skin surface of larval Acantlaorhodeus gracilis from Korea were observed. They spawned from late March through the middle of June. A fish spawned at least 5 times and the number of eggs averaged 304 per oviposition. Unfertilized eggs are nearly ovoid-shaped(mean$\pm$SD=$2.09{\pm}0.04\;mm$ in length with range of 2.05 to 2.13 mm and mean$\pm$SD=$1.26{\pm}0.02\;mm$ in breadth with range of 1.24 to 1.30 mm measured for the 50 eggs) and opaque yellow color. The tip of egg membrane at the animal pole side swelled and formed a few hilly projections. The shape of the eggs was just like a loquat. Most of embryos began to hatch out in thirty-eight hours after insemination at $22{\pm}1^{\circ}C$ in water temperature. As regards the morphological characters of the eggs and larval development, Acan. gracilis was similar to Acan. asmussi, Acheilognathus rhombeus, A. longipinnis and Pseudoperilampus typus. The larvae of this species is unique particularly in the following two characters, i.e., 1) scale-like tubercles ellipsoided in a diagonal cross section on the whole body and 2) incessant wiggly movement pattern as that of fly maggot, with the larvae of the above mentioned species. These characters seem to reflect the phylogenetic relationships among acheilognathine fishes. On the other hand, this species and Acan. asmussi are spring-summer spawning bitterlings. And also these species never retard the larval growth in such larval stage as the duration from Stage B to Stage D.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.79-92
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• 1992

This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

• Korean Journal of Soil Science and Fertilizer
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• v.10 no.3
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• pp.103-134
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• 1977

The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.