• Korean Journal of Veterinary Research
• /
• v.26 no.1
• /
• pp.117-124
• /
• 1986

The present study was undertaken to evaluate rosette formation and NK activity of splenic lymphocytes in 3-methylcholanthrene (MCA) treated mice. Mice were sensitized iv with 0.1ml of 1% sheep red blood cell (SRBC) suspension were treated with a single ip injection of olive oil alone or with different doses of MCA in oil at various time before or after sensitization, and were challenged at 4 days after SRBC. Rosette formation and NK activity of splenic lymphocytes were measured at 24 hours after challenge. Erythrocyte(E) rosette formation of splenic lymphocytes was significantly depressed in mouse treated with large dose of MCA (5~50mg) regardless of injecting time. But, there was no difference in the response between the treated with small dose of MCA (0.5mg). Whereas erythrocyte-antibody(EA) rosette or erythrocyte-antibody-complement(EAC) rosette forming cells were significantly depressed by MCA. Under small dose of MCA (0.5mg), any difference of NK activity was not observed in all course of injecting time. But, under large dose of MCA, the activity was markedly inhibited to about half the values seen in control and this suppression was transient, resulting that the normal level was reached again 19 days after MCA. These results, which conform with the predictions of immunosuppression hypothesis, suggest that MCA inhibits immunological responses including NK activity and thereby allows the outgrowth of antigenic neoplastic cells.

• Fisheries and Aquatic Sciences
• /
• v.5 no.3
• /
• pp.145-149
• /
• 2002

The effects of pH, temperature and various inhibitors on the proteolytic activity of the cell lysate of Uronema marium were investigated using colorimetric and substrate gel electro­phoretic methods. The cell lysate of U. marinum showed proteolytic activity over a wide range of pH, and pH optima ranged from pH 5 to 7. The proteolytic activity was increased according to a rise of temperature but decreased at $40^{\circ}$. The proteolytic activity of the parasite lysate was significantly inhibited by protease inhibitors including trans-epoxysuccinyl -L-leucylamido-(4-guanidino) butane (E-64), pepstatin A, phenyl-methanesulfonyl fluoride(PMSF), and ethylenediamine-tetraacetic acid (EDTA). Preincubation of the lysate with E-64 showed the maximum inhibition of the caseionolytic activity. Four protease bands (152, 97, 67 and 40 kDa) were detected by gelatin SDS-PAGE. Significant inhibition of caseinolytic activity and complete abolition of a 152 kDa band in gelatin SDS-PAGE by EDTA indicated that the cell lysate of U. marinum had a metalloprotease Another three proteolytic bands were inhibited by E64, a cysteine protease inhibitor. Preincubation of the cell lysate with pepstatin or PMSF had no effects on the protease bands.

• Fisheries and Aquatic Sciences
• /
• v.6 no.4
• /
• pp.180-186
• /
• 2003

The full-length cDNA encoding the pre-protein growth hormone (sfGH) from spotted flounder (Verasper variegatus) was amplified by the rapid amplification of cDNA ends (RACE) using degenerated oligonucleotide primers derived from conserved growth hormone sequences. It consists of 901 nucleotides in length, including the coding region of 609 nucleotides, 111 nucleotides of a 5' untranslated region, and 181 nucleotides of a 3' untranslated region. The conserved polyadenylation signal (AATAAA) lies 12 bases upstream from the poly (A) tail. The deduced amino acid sequence shows an open reading frame encoding a pre-protein of 203 amino acids and a putative signal peptide of 17 amino acids, suggesting that the mature hormone consists of 186 amino acids. The analyses of sfGH reveal some unique structural features. The repetitive sequences are located in the 5' untranslated region of sfGH cDNA and consist of tandem arrays of imperfect direct repeat monomers. Moreover, sfGH contains six Cys residues, as opposed to four or five in other GHs, and it is clearly distinguishable from olive flounder (Paralichthys olivaceus) GH, which lacks a region corresponding to residues 175-188 in alignment positions. It has important implications from an evolutionary standpoint, suggesting possible divergence among flatfishes.

• Fisheries and Aquatic Sciences
• /
• v.7 no.2
• /
• pp.58-63
• /
• 2004

Five urease-positive Vibrio parahaemolyticus strains were isolated from Kamak Bay in Yeosu in 2002 and 2003. V. parahaemolyticus YKB4 and YKB14 were isolated from seawater, YFB20 from black rockfish (Sebastes schlegeli), and YFO2l and YFO22 from olive flounder (Paralichthys olivaceus). The five urease-positive strains (YKB4, YKB14, YFB20, YFO21, and YFO22) did not show hemolysin and protease activity, while they did alter in color (to red) as the bacteria grew in the urea broth medium. All samples showed identical biochemical characteristics as a reference strain, V. parahaemolyticus KCTC2471, except in urease production. The five urease-positive strains showed urease activities at a mid stationary phase, and their activity was maximal in the late stationary phase of their culture supernatant. The addition of urea to the Luria-Bertani (LB) broth medium significantly affected the initial production of urease of V. parahaemolyticus isolates. Mortality by urease-positive V. parahaemolyticus YKB4, YKB14, YFO2l, and YFO22 was significantly high, being$60-80\%$, while YFB20 only reflected a rate of $20\%$. Protease-positive V. parahaemolyticus FM39 and FM50 showed a $40\%$ and $60\%$ mortality rate, respectively. However, hemolysin-positive V. parahaemolyticus had no mortality, like the non-pathogenic V. parahaemolyticus KCTC2471, while V. vulnificus resulted in a $40\%$ mortality rate. Injection with urease-positive V. parahaemolyticus strains showed mortality within 12 hrs in mice, and the strains could be isolated from the dead mice.

• Korean Journal of Microbiology
• /
• v.41 no.2
• /
• pp.117-124
• /
• 2005

Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

• KSBB Journal
• /
• v.27 no.2
• /
• pp.91-96
• /
• 2012

In this study, a newly synthesized thiazolidinedione derivative, TD49 with a highly selective algicide to red tide, was examined in order to evaluate the bioconcentration on aquatic organisms of coast. BAF (accumulation of TD49 by aquatic food chain) and BCF (accumulation of TD49 by sea water) were examined employing the shrimp (Fenneropenaeus chinensis) as the feed organism, and the olive flounder Paralichthys olivaceus as a consumer in marine ecosystem. Bioconcentration degree in sea water showed that the order in P. olivaceus was viscera > gill > muscle. The average BCF values of TD49 were 67.70, 63.32 and 20.25 at viscera, gill and muscle, respectively. Bioaccumulation degree using feed showed that the order in the organs of P. olivaceus was viscera > gill > muscle. The average BAF values of TD49 were 175.89, 114.88 and 32.59 at viscera, gill and muscle, respectively. When compared with two results, the accumulation by the food and water was higher than that by water. After the elimination experiment in sea water, the TD49 concentration was 2.81 nmole/g in the viscera and were not found in the gill and the muscle. More than 50% of the accumulated TD49 were eliminated from viscera in 7 days and all the accumulated TD49 were eliminated from gill and muscle in 7 days. On the other hand, the octanol/water partition coefficient (log $K_{ow}$) was measured to be 3.66 and experimental BCF of this study was 67.7.

• Journal of Aquaculture
• /
• v.16 no.2
• /
• pp.118-123
• /
• 2003

This study was conducted to investigate the disinfection effects of chlorine dioxide ($ClO_2$) on 4 fish pathogenic bacteria (Vibrio anguillarum, Edwardsiella tarda, Streptococcus sp. and Staphylococcus sp.) isolated from infected olive flounders. The bacteria were exposed to different concentrations of ClO$_2$ (0.129, 0.246 and 0.455 ppm) and response times (0.5, 1, 3, 5 and 10 min), and then were incubated for 12 hr. The effective disinfection concentrations of $ClO_2$ against experimental bacteria by $ClO_2$ for 0.5 min were observed with 0.455 ppm for Staphylococcus sp., 0.246 ppm for V. anguillarum and E. tarda, and 0.129 ppm for Streptococcus sp., respectively. The duration of exposure at low concentration of $ClO_2$ increased for the disinfectant ability to experimental bacteria.

• Journal of Environmental Health Sciences
• /
• v.29 no.2
• /
• pp.80-86
• /
• 2003

To evaluate the effect of cyclohexane(CH) treatment on the serum levels of glutathion S-transferase(GST) activity in liver damaged animals, damaged liver was induced with pretreatment of 50% $CCl_4$ dissolved in olive oil (0.1 m1/100g body weight) intraperitoneally 17 times every other day. To $CCl_4$-treated rats, CH (1.56 g/kg body weight, i.p) was injected once and then the animals were sacrificed at 4 hours after injection of CH. The $CCl_4$-treated animals were identified as severe liver damage on the basis of liver functional findings, 1,e, increased serum levels of alanine aminotransferase(ALT), alkaline phosphate(ALP) and xanthine oxidase(XO) activities. On the other hand, $CCl_4$-treated animals injected with CH once($CCl_4$-pretreated animals) showed more decreased serum levels of ALT and XO, and more increased those of ALP rather than $CCl_4$-treated animals. In case of comparing the GST with ALT activity in liver, both $CCl_4$-treated and pretreated animals showed similar changing pattern of enzyme actvity. Especially $CCl_4$-pretreated animals showed significantly increased serum level of GST actvity compared with the $CCl_4$-treated those, whereas those of ALT showed reversed tendency. In aspects of GST enzyme kinetics, $CCl_4$-pretreated animals showed higher Vmax of liver GST enzyme than $CCl_4$-treated animals. In conclusion, injection of CH to the liver damaged rats led to enhanced liver damage and more increased activity of serum GST which may be chiefly caused by the enzyme induction.

• Biomedical Science Letters
• /
• v.12 no.4
• /
• pp.361-370
• /
• 2006

To evaluate an effect of pathological liver damage on the conjugation of cyclohexane metabolites, rats were pretreated with 50% $CCl_4$ dissolved in olive oil (0.1 ml/100 g body weight) 10 or 17 times intraperitoneally at intervals of every other day. On the basis of liver function, the animals pretreated with $CCl_4$ 10 times were identified as acutely liver damaged ones and the animals pretreated with $CCl_4$ 17 times were identified as severly liver damaged ones. To these liver damaged animals, cyclohexane (a single dose of 1.56 g/kg body weight, i.p.) was administered at 48 hr after the last injection of $CCl_4$. The rats were sacrificed at 4 or 8 hr after injection of cyclohexane. The cyclohexane metabolites, cyclohexanol (CH-ol), cyclohexane-1,2-diol (CH-1,2-diol), cyclohexane-1,4-diol (CH-1,4-diol), and their glucuronyl conjugates and cyclohexanone were detected in the urine of cyclohexane treated rats. The urinary concentration of cyclohexane metabolites was generally more increased in liver damaged animals than normal ones, and the increasing rate was higher in $CCl_4$ 17 times injected rats than 10 times injected ones. And liver damaged.ats, especially $CCl_4$ 17 times treated ones, had an enhanced ability of glucuronyl conjugation to CH-ol analogues compared with normal group. Futhermore, CH-1,2 and 1,4-diol were all conjugated with glucuronic acid in $CCl_4$ 17 times injected animals. On the other hand, the increasing rate of activities of hepatic cytochrome P450 dependent aniline hydroxylase, alcohol dehydrogenase and urine diphosphate glucuronyl transferase was higher in 17 times $CCl_4$-treated rats compared with normal and $CCl_4$ 10 times injected animals. Taken all together, it is assumed that an increased urinary excretion amount of cyclohexane metabolites in liver damaged rats might be caused by an increase in the activities of cyclohexane metabolizing enzymes. And enhanced conjugating ability of CH-ol in liver damaged animals and novel finding of conjugating form of CH-1,2 and 1,4-diol might be caused by increase in the activity of hepatic diphosphouridine glucuronyltransferase.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.39 no.6
• /
• pp.454-459
• /
• 2006

A simultaneous high-performance liquid chromatography assay method for amoxicillin and ampicillin in fish products was developed, evaluated, and validated by monitoring these antibiotics in fish samples obtained from aquaculture and distribution. The recovery rate of this method was higher than those of conventional methods and was 95.3-106.6% for amoxicillin and 81.4-92.4% for ampicillin. Our pretreatment procedure sufficiently removed or reduced materials affecting HPLC analysis, such as low-molecular-weight substances. The performance limit of this method was evaluated as 0.01 ppm of amoxicillin and ampicillin in fish muscle. Finally, 171 fish samples, including olive flounder (Paralichthys olivaceus), common sea bass (Lateolabrax japonicus), and black rock fish (Sebastes schlegeli) collected from fish farms in the coastal area between April and September 2005 were analyzed to evaluate the overall efficiency of the method and to monitor the actual of amoxicillin and ampicillin usage in fish farms. The results indicated that the developed method was suitable for analyzing amoxicillin and ampicillin in fish muscle, and determined that those antibiotics were being used for fish farming but were not detected in fish samples during the shipping and distribution stages.