Yong-Jae Han

Innate immune response and disease resistance in Carassius auratus by triherbal solvent extracts

This study reports the effect of aqueous, ethanol and methanol triherbal solvent extract from Azadirachta indica, Ocimum sanctum and Curcuma longa on innate immune mechanisms such as phagocytosis activity, respiratory burst activity, alternative complement activity and lysozyme activity and disease resistance in goldfish (Carassius auratus) against Aeromonas hydrophila. Fish were intraperitoneally injected with different doses of 0, 5, 50 and 100 mg kg(-1) body weight of each triherbal solvent extracts. The functional immunity in terms of percentage mortality and Relative Percent Survival (RPS) and innate immune response was assessed on week 1, 2 and 4 by challenging with live A. hydrophila (1 x 10(7) cells ml(-1)). All the chosen innate immune parameters were enhanced in the ethanol and methanol triherbal solvent extract treatment after week 2. However, the aqueous triherbal extract was enhanced only after week 4. The ethanol and methanol triherbal solvent extracts administration preceding the challenge with live A. hydrophila decreased the percentage mortality in the experimental groups with the consequence increase in RPS values. The study indicates that all the doses of ethanol or methanol triberbal solvent extracts could be positively influence the immune response and protect the heath status of goldfish against A. hydrophila infection.

Effect of Punica granatum solvent extracts on immune system and disease resistance in Paralichthys olivaceus against lymphocystis disease virus (LDV).

We report the effect of aqueous, ethanol, and methanol solvent leaf extracts of Punica granatum on innate immune mechanisms, such as phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity and functional immunity in terms of percentage cumulative mortality and Relative Percent Survival (RPS) in olive flounder Paralichthys olivaceus naturally infected with lymphocystis disease virus (LDV) after 8 weeks. Infected fish were intraperitoneally administered with 0, 5, 50, and 100 mg kg(-1) body weight of solvent extracts. In groups treated with 50 and 100 mg kg(-1) body weight, the chosen innate immune parameters significantly increased after 8 weeks when compared to 0 mg kg(-1) dose, but not with 5 mg kg(-1). Administration of P. granatum solvent extracts for 8 weeks significantly reduced the percentage mortality with the consequent increase in RPS. The results suggest that intraperitoneal administration of the leaf extracts of P. granatum at 50 or 100 mg kg(-1) dose clearly enhance the innate immune responses and disease resistance after 8 weeks in P. olivaceus against natural LDV infection.

Effect of traditional Korean medicinal (TKM) triherbal extract on the innate immune system and disease resistance in Paralichthys olivaceus against Uronema marinum

We report the effect of aqueous-, ethanol- and methanol-solvent-derived extracts of three traditional Korean herbs, Punica granatum, Chrysanthemum cinerariaefolium and Zanthoxylum schinifolium, by monitoring the innate immune mechanisms, such as phagocytosis activity, respiratory burst activity, alternative complement activity and lysozyme activity and the functional immunity in terms of percentage mortality and relative percent survival (RPS) in olive flounder (Paralichthys olivaceus) against Uronema marinum (1 x 10(5)ciliates ml(-1)) for 30 days. Fish were intraperitoneally administered with 5, 50 and 100 mg kg(-1) body weight of each traditional Korean medicinal (TKM) solvent extract except the control and infected untreated groups. In all the treated groups at concentrations of 50 and 100 mg kg(-1) body weight, the chosen innate immune parameters were found significantly enhanced when compared to 0 mg kg(-1) dose. However, at 5 mg kg(-1) the tested immune parameters did not vary. Administration of TKM solvent extracts preceding the challenge with U. marinum for 30 days significantly reduced the percentage mortality with the consequent increase in RPS. Administration of 50 and 100 mg kg(-1) TKM solvent extracts clearly enhanced the innate immune responses and disease resistance in P. olivaceus against U. marinum.

Immunomodulatory effect of sodium alginate enriched diet in kelp grouper Epinephelus brneus against Streptococcus iniae.

The effect of diets containing sodium alginate at 0, 0.5, 1.0, and 2.0 g kg⁻¹ following challenge with Streptococcus iniae in kelp grouper Epinephelus bruneus were assessed with reference to survival rate and innate immune parameters such as alternative complement, lysozyme, natural haemagglutination, respiratory burst, superoxide dismutase, and phagocytic activities on week 1, 2, and 4. Fish fed with sodium alginate containing diet at 1.0 and 2.0 g kg⁻¹ after being challenged with S. iniae had higher survival rates of 75% and 60%, respectively than those fed with control diet (0 g kg⁻¹). With any enriched diet the percentage of macrophages significantly decreased from week 1-4, while the percentage of neutrophils and lymphocytes significantly increased. The alternate complement activity, natural haemagglutination, and phagocytic activities of infected fish fed with sodium alginate containing diet at 1.0 g kg⁻¹ on week 2 and 1.0 and 2.0 g kg⁻¹ diets on week 4 were significantly higher when compared to the control. The lysozyme, respiratory bursts, and superoxide dismutase activities of fish fed with enriched diets at 1.0 and 2.0 g kg⁻¹ were significantly increased on week 2 and 4. We therefore recommend that at 1.0 or 2.0 g kg⁻¹ dietary administration of sodium alginate can enhance innate immunity and disease resistance in kelp grouper against S. iniae.

Effect of a mixed herb-enriched diet on the innate immune response and disease resistance of Paralichthys olivaceus against Philasterides dicentrarchi infection.

We investigated the effect of a mixed herb-enriched diet obtained from pomegranate Punica granatum, Dalmatian chrysanthemum Chrysanthemum cinerariaefolium, and mastic-leaved prickly-ash Zanthoxylum schinifolium on innate immune mechanisms (e.g., phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity, and disease resistance) of olive flounder Paralichthys olivaceus against the scuticociliate Philasterides dicentrarchi. All experimental groups were challenged with P. dicentrarchi (1 x 10(5) ciliates/mL) through intraperitoneal administration of the pathogen (50 microL) on day 1. On day 7, the infected groups were fed 0, 5, 50, and 100 mg/kg of the enriched diets. The innate immune parameters, cumulative mortality, and relative percent survival (RPS) were assessed at weeks 1, 2, 3, and 4. Administration of 50 or 100 mg/kg of the herbal-enriched diet enhanced immunity throughout the experimental period. However, at the 5-mg/kg dosage, the enriched diet did not enhance the innate immune estimates at any time. At doses of 50 and 100 mg/kg, administration of the diet preceding the challenge with P. dicentrarchi decreased the percentage cumulative mortality in the experimental groups and thereby increased RPS values. This study reports that administration of 50 or 100 mg/kg mixed herbal-enriched diet could positively influence the innate immune response to P. dicentrarchi and enhance the health status of olive flounder with respect to this microbe.

Physico-Chemical Characteristics of Aquacultural Discharging Water in Jeju Island

Physical and chemical analyses of water discharged from 4 crowded farms (Sungsan, Pyosun, Wimi and Daejung) in Jeju island were performed from July, 2006 to Dec, 2006, and the result of the analyses showed that hydrogen ion concentrations (pH) for water discharged from Sungsan farm was 7.74, Pyosun was 7.68, Wimi was 7.68 and Daejung was 7.7. Salinity levels for Sungsan, Pyosun and Wimi had an average of 3133 / indicating characteristics of far distance areas, whereas that of Daejung was 28.81 %, which was far lower compared to regular sea water salinity. As the result of measuring dissolved oxygen (DO) for each area, each area showed first graded DO for each discharged water based on water quality level for each sea district. The result of measuring the temperature for discharged water showed that water temperatures for summer were 2325C, and those for winter were 16 18C. Nitrogen concentrations for discharged water exceeded each sea area`s water quality level in all farms. In the case of phosphate, its average value was 0.48 mg/l for Sungsan, 0.55 mg/I for Pyosun, 0.66 mg/I for Wimi, and 0.44 mg/l for Daejung, and chemical oxygen demand (COD) was shown to be 1.5 mg/l 1.8 mg/I, 1.6 mg/I and 2.3 mg/I for Sungsan, Pyosun, Wimi and Daejung respectively. For suspended solids (SS), the average concentration was 19.3 mg/I, 21.2 mg/I, 21.3 mg/I and 18.5 mg/I for Sungsan, Pyosun, Wimi and Daejung respectively. The results of physical and chemical analyses for discharged water in farms based on time showed that almost all items were shown to increase in the forenoon and decrease, overall, in the afternoon.

Microbiological Pollution Investigation of Lowland Spring Water at the Jeju City Waterfront, Jeju Island

Bacteriological examination of spring water in Jeju City was conducted. A total of 21 spring water samples were collected from January to April, 2010. During the study period, the range of temperature was 0.6 to 15.4℃, and the results of the analyses showed that hydrogen ion concentrations (pH) for spring water were 0.43 to 7.9. Salinity levels for the samples averaged from 3.0 to 1.64%, and levels of water-dissolved oxygen were 1.85 to 6.06 mg/l. The range of total coliforms in spring water samples at 21 stations located in the designated spring water were 1,600 MPN/100 ml. Furthermore, the range of geometric means of total coliforms was 9.9-151.6 MPN/100 ml, while the range of fecal coliforms in spring water samples at 21 stations located in the designated spring water area was 1,600 MPN/100 ml. Finally, the range of geometric mean of fecal coliforms was 3.1-151.6 MPN/100 ml. The level of microbial contamination was examined in 21 samples for indications of bacterial contamination such as heterotrophic bacteria, Escherichia coli O157:H7, Salmonella spp., Vibrio parahaemolyticus, Staphylococcus aureus, and Shigella spp. were frequently detected in the spring water. Salmonella spp., Shigella spp., Vibrio parahaemolyticus, and S. aureus were detected in the range of 0-0.5×10 1 , 0-0.1×10 1 , 0-0.1×10 1 , and 0-0.3×10 1 CFU/ml, respectively, while E. coli O157:H7 was not detected in the examined spring water samples.

Identification of Microorganisms Isolated from Jeju Coastal Sedimentary Layer and Characterization of Their Proteases.

In this study, protease-producing bacteria were isolated from the marine sedimentary layer in coastal Jeju. We isolated 2 protease producing strains (SK-2 and SK-125) and tested their protesase producing activities. Gram staining and BIOLOG of isolated strains revealed that strains SK-2 and SK-125 belong to Bacillus and Pseudoalteromonas families, respectively. The 16S rDNA nucleotide sequences analyses of the isolated strains showed 99% sequence homology with those of Bacillus sp. and Pseudoalteromonas sp.; therefore, the isolated strains SK-2 and SK-125 were named Bacillus sp. SK-2 and Pseudoalteromonas sp. SK-125, respectively. The optimum conditions for the cell growth of protease activities were obtained when the both isolates were cultured at 30℃, 96 hrs and pH 7～8.