• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.71-73
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• 2000

Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.339-347
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• 2000

An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.

• Journal of Chest Surgery
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• v.36 no.8
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• pp.545-558
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• 2003

Adult respiratory distress syndrome (ARDS) is of particular interest because of its severity of the associated lung injury and its high mortality. However, the pathophysiologies of ARDS in infant and childhood groups are still not well clarified inspite of many previous investigations. To investigate the time course of pathophysiology of ARDS in infant and childhood groups, this study was designed with experimental endotoxin-induced ARDS model using young rabbits (8 week-old). Material and Method: Rabbits were divided into the control group (n=8) and the endotoxin-treated group (n=32). The endotoxin group was subdivided into 4 groups by the sampling times as 3, 6, 12 and 24 hr-groups (G- $E_{3,6,12,24,}$ each n=8). The experimental ARDS was made by a bolus injection of endotoxin (Escherichia coli serotype 055 : B5, 0.50 mg/kg) via rabbit ear vein. For evaluation of the hematologic and inflammatory markers, and superoxide dismutase (SOD) concentrations, the blood samples were taken from the heart. The bronchoalveolar lavage fluid (BALF) were obtained for analysis of the leukocytes and protein concentration. With biopsy of the lung, histopathologic changes of the lung were also evaluated. Result: In the endotoxin groups, significant leukopenia (owing to pancytopenia) occurred in 3 and 6-hr groups, which was followed by significant leukocytosis (owing to neutrophilia) in the 12 and 24-hr groups (p<0.05). Serum levels of tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1 $\beta$ (IL-1 $\beta$) in the endotoxin groups were higher than those of control group (p<0.05). Serum levels of superoxide dismutase (SOD) of G- $E_{3}$ and G- $E_{6}$ were higher than those of control group, whereas those of G- $E_{12}$ were lower than those of control groups (p<0.05). Total leukocyte counts and protein con-centrations in BALF were significantly elevated in the endotoxin groups compared to the control group (p < 0.05). The hemorrhagic pattern of BALF showed occurred in the endotoxin groups. The endotoxin groups (in G- $E_{6}$) had severe infiltration of inflammatory cells (lymphocyte and monocyte) in the pulmonary interstitium and parenchyma, migrations of neutrophil and eosinophil into alveolar spaces and interstitial widening, which are the evidences of acute lung injury. In the endotoxin groups, there were significant positive correlations between the BALF findings and the immunologic markers (TNF-$\alpha$, IL-1$\beta$, SOD) (p<0.05). Conclusion: Severe acute lung injury occurred in all the endotoxin-treated rabbits. The pathophysiologic findings were so progressive until 6-hr by time dependant pattern, and then recovered slowly, Variable hematologic, immuno-logic, and pathologic factors were well correlated in the development and progression of endoxin-induced lung injury. The pathophysiologic responses were sensitive and rapid in young rabbit Young rabbit seemed to be a useful experimental animal model for infant and childhood groups.roups.

• The Journal of Dong Guk Oriental Medicine
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• v.7 no.2
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• pp.121-140
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• 1999

The various oriental herbal medicines, which have usually been used for treatment of reducing fever, purging intense heat and detoxication, were screened to determine the antibacterial activity and the minimal inhibitory concentration against pulmonary disease-causing Klebsiella pneumoniae, Streptococcus pyogenes, and Streptococcus pneumoniae. The results obtained were as follows: 1. Among the 23 oriental medicines tested, the water-soluble extracts of Coptis japonica, Scutellaria baicalensis and Picrorrhiza kurrooa showed the antibacterial activity against K.pneumoniae and that of C. japonica against S. pyogenes. The antibacterial activities of C. japonica, Prunusmume, Schizandra chinesis, Scutellaria baicalensis were also found against S.pneumoniae. When C.japonica was used, the high antibacterial activity was shown against Bacillus subtilis and other extracts showed a little activity against B. subtilis and E. coli as a control. 2. The ethanol-soluble extracts of Patrinia scabriosaefolia, P. mume, S. baicalensis, S. chinesis showed the antibacterial activity against K. pneumoniae and those of S. baicalensis, C. japonica, S. chinesis, P. mume agaist S. pyogenes and S. pnuemoniae. However, those extract showed a little antibacterial activity against B. subtilis and E. coli except for that the extract of C. japonica showed comparatively high growth inhibition of B. subtilis. 3. Among the medicinal herbs tested, the water and ethanol extrats of C. japonica showed very extcellent antibacterial activity against the pathogenic bacteria and controls. 4. When the water-soluble extracts of C. japonica and S. baicalensis, minimal inhibitory concentrations (MICs) against K. pneumoniae were $10mg/m{\ell}$ and $22 mg/m{\ell}$, respectively. The MICs of the ethanol-soluble extracts of P. mume and P. scabriosaefolia were $5mg/m{\ell}$ and $20mg/m{\ell}$, respectively. 5. For the MICs against S.pyogenes, C. japonica showed $15mg/m{\ell}$ with the water-soluble extract and P. mume and C. japonica with the ethanol-souble extract did $5mg/m{\ell}$ and $10mg/m{\ell}$, respectively. 6. For the MICs against S. pneumoniae, C. japonica and P. mume with the water- and ethanol-souble extract showed $5mg/m{\ell}$ and $10mg/m{\ell}$, respectively. As a result, the highest antibacterial activity was found in the water- and ethanol-soluble extracts of C. japonica against pulmonary disease-causing bacteria, K. pneumoniae, S. pyogenes. and S. pnuemoniae. Also, the water- and ethanol-soluble extracts of S.chinesis. P.mume, S.baicalensis, and P.kurrooa showed hight antibacterial activities.

• Journal of Veterinary Clinics
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• v.10 no.2
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• pp.171-184
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• 1993

In Seoul area, there are so many kennel clubs, veterinary hospital, pet establishment and breeding confers that various problems have occurred. They are crowed pet houses, poor sanitation, stress to puppies, sudden environmental changes to puppies and unvaccination against parvovirus, canine distemper virus, parainfluenza virus, infectious hepatitis caused by Adenovirus type I and Leptospira. Several studies were made to survey the infectious agents involved in acute diarrhea of poppies in Seoul are, such as history taking, physical examination, complete blood count and serum chemistry, histopathological finding, bacterial isolation and identification, and hemagglutination test in feces and hemagglutination inhibition test in serum against parvovirus, respectively. The results obtained are summarized as followed. 1. The percent of PCV (30.5$\pm$5.6) and concentration of Total protein(5.0$\pm$0.8) resulted from statistical analysis are significantly lower than normal values (p<0.05), respectively. In addition, fibrinogen (505$\pm$326) was significantly higher an normal value (p<0.001), Band neutrophil (22.9$\pm$12.7) showed signifiant difference (P<0.01). decreased monocyte (3.2$\pm$2.1) and eosinophil (0.7$\pm$0.8) values appeared statistically significant (p<0.001), lymphocyte (16.7$\pm$9), as well. 2. The concentrations of calcium .(8.0$\pm$2.8), glucose (40.1$\pm$31.4), and albumin (2.0$\pm$0.39) were lower than normal values (p<0.01, p<0.01, p<0.001), respectively. Also Inorganic phosphate (7.1$\pm$2.4), pH (p<7.9f:0.2), and Blood Urea Nitrogen (40.41=37.1) were significantly higher than normal values (p<0.001, p(0.001, p<0.05), respectively. 3. Simple and mixed infections occupied 18% and 82% in the distribution of causes in puppies with acute diarrhea, respectively. 4. As puppies got older, incidence of acute diarrhea caused by Staphylococcus aureus was decreased to 13% and infection of canine distemper virus was increased to 53%, but E coli and canine parvovirus always showed high frequency of outbreak in the body weight ranged from 35g to 7.8Kg. 5. As showed in table 5, infections of E coli and Canine Parvovirus showed high outbreak regardless of the age which is classified into three stages, 35~50 day, 60 day and 75~day to 1 year, canine distemper virus appeared increased, but in case of Staphylococcus aureus, visa versa. 6. In comparison wi methods for the laboratory diagnosis diagnosis parvovirus, Hemagglutination test showed positive reaction in 25% and mean serum antibody titer measured by Hemagglutination inhibition test showed 2779 (n=20). In addition, positive reaction was 90% (18/20). 7. In histopathological studies, enteritic and pneumonic lesions were indicated in 53.7% and 39.5% of samples, respectively. 8. Etiologic diagnosis based on the history taking, clinical signs and histopathological findings in puppies with acute diarrhea and vomiting indicated that 50% of puppies were infected by canine parvovirus and distemper virus. 9. In the parasitological examination made by simple flotation with saturated zinc sulfate tour parasites found were Isospora canis, Toxocara canis, Ancylostoma spp and Toxocara spp. Isospora canis and Toxocara canis were more frequently found among those parasites of diarrhoeic causes in puppies ranged from 35 days to 1 year. Their infestation rates were 15% and 13% respectively.

• Korean Journal of Poultry Science
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• v.35 no.1
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• pp.71-78
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• 2008

This study was conducted to investigate the effects of dietary supplementation of herbs and plant extracts (PE) on the performance, small intestinal microflora and immune response in laying hens. A total of 1,440 Hy-Line Brown laying hens of 67 wks old were assigned to one of the following 9 dietary treatments : T1 : Control (C), T2 : C + Avilamycine 6 ppm, T3 : C + Herb $Mix^{(R)}$ 0.2%, T4 : C + Biostrong $510^{(R)}$ 0.02%, T5 : C + $APEX^{(R)}$ 0.02%, T6 : C + $Digestarom^{(R)}$ 0.02%, T7 : C + $Phellozyme^{(R)}$ 0.1%, T8 : C + $Galicin^{(R)}$ 0.05%, T9 : C + CRINA $Poultry^{(R)}$ 0.05%. Each treatment was replicated 8 times with twenty birds housed in 2 bird cages. Twenty bird units were arranged according to completely randomized block design. Feeding trial lasted 6 wks under 16 hours lighting regimen. Hen-day egg production was not significantly different among the treatments, but that of supplemented groups tended to be higher than the control. There were significant differences among treatments in feed intake and feed conversion ratio. Feed intake was higher in the supplemented groups than the control. Feed conversion ratio was higher in T8 than other treatments. Egg shell color index and egg yolk color index were significantly different among treatments. Egg shell color was affected by supplements and egg yolk color index of T9 (PE-CRINA) was significantly higher than the control. Haugh unit was not significantly different among treatments. There were significant differences in leukocytes and erythrocytes parameters. The level of serum WBC and stress index (heterophil/lymphocyte) were higher in supplemented groups than the control. The level of RBC tended to be lower in the herb or PE groups than the control. The concentration of serum IgG was not significantly different among the treatments, but all those of the supplemented groups were higher than the control. The number of Lactobacilli spp. tended to increase and that of Cl. perfrigens decrease in the supplemented groups. The number of E. coli significantly decreased in the supplemented groups. The results of this experiment showed that feeding herbs and plant extracts to laying hens tended to improve the egg production and affect positively on the level of blood parameters and small intestinal microflora.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.479-486
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• 2006

The current study was conducted to examine the effect of feeding and management practices on milk quality and dairy farm productivity in Korea. Fifty dairy farms in Gyunggi (11), Gangwon (22), Chungnam (17) provinces were surveyed to collect data on the herd size, housing style, feeding management, waste disposal, milking practices and milk yield. Milk tank samples from all farms under study were also collected to enumerate its composition and quality parameters. Large dairy herds are equiped with better housing, milking and waste control facilities than medium and small dairy herds. Higher concentrate feeding to lactating cows was noticed in small dairy herds (47.51 %) than in medium (32.59 %) and large dairy herds (31.82 %). The decrease in concentrate feeding to lactating cows with increase in number of cows per farm resulted in a simultaneous increase in the use of imported forages. Bacterial count in milk was affected by housing and milking facilities at dairy farms. Higher bacterial counts (Coliform and E. coli) in milk were observed in cows housed in stanchion than those under free stall with saw dust bedding. The bacterial counts were higher with bucket milking system than with pipe-line and parlour systems. The increase in the number of dairy cows per farm and thus better management and milking facilities resulted in a reduction in somatic cell score. Milk yield (per cow) was higher in herds with less somatic cell score. Average milk protein concentration was between 2.89 to 2.98 % and milk urea nitrogen was between 21.81 to 23.31mg/ml on surveyed dairy farms. This study concluded that large herd size with better dairy cow management facilities is crucial to produce quality milk with better dairy farm income.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.687-700
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• 2004

This experiment was carried out to examine the fennentation properties of yogurts with or without Lyeii fructus, Lyeii folium and Lyeii cortex extract as additives at concentrations of 1.0%. The effects on promoting the fermentation by Lycii fructus, Lycii folium and Lycii cortex additives were higher and pH was below 4.06 when Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus cosei, and Lactobacillus acidophilus, Bifidobacterium longum and Streptococcus salivarius ssp. thermophilus were used. The acid production was higher when S. salivarius ssp. thermophilus and L. delbrueckii ssp. bulgaricw were used. The average lactic acid bacteria counts was 2.62 ${\times}$ $10^9$ cfu/ml in the yogurt added with Lycii fructus extract and fermentation with S. salivarius ssp. thermophilus and L. delbrueckii ssp. bulgaricw. The lactose hydrolysis ratio was higher in the milk added with Lycii fructus extract(36.11%), Lycii folium extract(37.76%) and Lycii cortex extract(32.70%) when S. salivarius ssp. thermophilus and L. delbrueckii ssp. bulgaricus were used. The isobutylic acid concentration was(34.39 to 37.72 mM) with S. salivarius ssp. thermophilus and L. delbrueckii ssp. bulgaricus. The viscosity of yogurt was 1,615 to 2,030 cP in yogurts added with skim milk and L. acidophilus; B. longum and S. salivarius ssp. thermophilus were used. The sensory scores of colors, tastes and overall acceptability of yogurt with Lycii cortex extract were shown 3.34 to 3.77 when fermented by L. cosei, L. acidophilus, B. longum and S. salivarius ssp. thermophilus. The cholesterol reducing effects were 17.38${\sim}$32.08% in all the yogurts and especially, greater effect(25.75 to 32.08%) for yogurts fermented with L. acidophilus KCTC3150 and L. salivarius subsp. salivarius CNU27. The inhibitory effects on the pathogenic bacteria by lactic acid bacteria added with Lycii fructus, Lycii folium and Lyeii cortex lower on S. typhimurium M-15, but higher on E. coli KCTC1021 and L. monocytogenes.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.259-268
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• 2003

Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinoma cells and is known to prevent symptomatic rotavirus infections. In this study, we tried to transfer the human lactadherin gene to the Chinese Hamster Ovary (CHO) cells using retrovirus vector system and tested inducible expression of the gene under the tetracycline-controllable promoter. At first, tetracycline-mediated inducibility was tested using E.coli LacZ marker gene. NIH3T3 cells co-infected with RevTet-On and RevTRE-LacZ retrovirus vectors showed that the cells responded to doxycycline (a derivative of tetracycline) in a dose-dependent manner, and prominent induction of the lacZ gene expression was observed from 1 $\mu\textrm{g}$/ml of doxycycline concentration. Based on the results of the pilot experiment, inductional expression of the human lactadherin gene was conducted using RevTet-On and RevTRE-Ltd retrovirus vectors. Analysis with the RT-PCR demonstrated successful inductional expression of the lactadherin gene in the Chinese Hamster Ovary (CHO) cells. Considering that constitutive overexpression of the exogenous genes in the target cells causes serious physiological imbalance, the results obtained in this study will be very useful especially in the studies of gene therapy and transgenic animal production.

• Journal of the Korean Applied Science and Technology
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• v.30 no.3
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• pp.400-410
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• 2013

This study was carried out to investigate the effects of feeding the broilers that are exposed to extreme heat stress by control of inverse lighting times with night restricted feeding of extreme heat diet(EHD1, 2: extreme heat diet) containing different amount of soy oil, molasses, amino acids and vitamin C on short chain fatty acid and blood lipid profile. 300 broiler chickens(Abaica strain) were randomized into four dietary treatment groups according to a randomized block design on the day they were hatched. The four dietary treatment groups were: T1(EHD 1, 10:00~19:00 Dark, 19:00~10:00 Light), T2(EHD 2, 10:00~19:00 Dark, 19:00~10:00 Light), T3(EHD 1, 09:00~18:00 Dark, 18:00~09:00 Light), T4(EHD 2, 09:00~18:00 Dark, 18:00~09:00 Light). The body weight gain of the broilers was highest in T2, and high in order T1, T4, T3(p<0.05). Weights of the lymphoid organ, thymus and bursa of Fabricius were high in T1, T2 as compared to T3, T4 but spleen was lower in T4 than T1, T2, T3(p<0.05). Blood triglyceride, total cholesterol and glucose were higher in T1, T2 than T3, T4(p<0.05). LDL-C was high in orderT4, T3, T2, T1 but HDL-C showed the opposite trend(p<0.05). Blood concentrations of IgG, IgG and IgM were higher in T1, T2 than inT3, T4, but the corticosterone concentration decreased significantly in them. In T1 and T2, Lactobacillus in the feces increased, but total aerobic bacteria, E.coli, coliform bacteria was decreased rather significantly, compared with those in T3 and T4(p<0.05). Concentrations of acetic acid, propionic acid and total SCFA in cecum were high in order T2, T1, T3, T4, but butyric acid, isobutyric acid, valeric acid, isovaleric acid were lower in T1, T2 than in T3, T4 (p<0.05).