Search

To identify potent plant extracts with strong antioxidant activity, we evaluated the free radical scavenging activity of 184 plant extracts obtained from the Freshwater Bioresources Culture Collection (FBCC) of Nakdonggang National Institute of Biological Resources (Republic of Korea), as various plant extracts have been used therapeutically to prevent chronic diseases associated with oxidative stress. From them, three plant extracts (FBCC-EP858 from Ammannia multiflora, FBCC-EP920 from Ammannia coccinea, and FBCC-EP1014 from Salix gracilistyla) were selected based on their abilities to scavenge the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical with more than 80% efficiency. We found that these extracts had in vitro half maximal inhibitory concentration (IC50) values ranging from 11.89 to 14.26 μg/mL and strong total antioxidant activity (corresponding to approximately 0.18, 0.22, and 0.23 mM Trolox, respectively). We also studied the effect of these extracts on RAW 264.7 macrophages and found that FBCC-EP920 significantly downregulated relative cell viability at a concentration of 100 μg/mL. However, the other two extracts, FBCC-EP858 and FBCC-EP1014, did not affect cell viability at the same concentration. Additionally, all three extracts inhibited hydrogen peroxide (H2O2)-induced reactive oxygen species (ROS) production and depolarization of mitochondrial membrane potential in RAW 264.7 macrophages. An additional experiment in zebrafish larvae showed that the three extracts reduced 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) fluorescent intensity induced by H2O2. The extracts also upregulated the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression, and an HO-1 inhibitor, zinc protoporphyrin (ZnPP), attenuated the extract-induced antioxidant activity both in vivo and in vitro. Taken together, these findings suggest that the extracts from A. multiflora, A. coccinea, and S. gracilistyla have potential free radical scavenging and antioxidant capacities both in vivo and in vitro by activating the Nrf2/HO-1 signaling pathway. These results could be useful for the prevention and treatment of various oxidative stress-mediated human diseases.

Sulfated polysaccharides extracted from brown algae are unique algal polysaccharides and potential ingredients in the cosmeceutical, functional food, and pharmaceutical industries. Therefore, the present study evaluated the cosmeceutical effects, including antioxidant, anti-wrinkle, anti-inflammation, and photoprotective activities, of

Airborne particulate matter (PM) originating from industrial processes is a major threat to the environment and health in East Asia. PM can cause asthma, collateral lung tissue damage, oxidative stress, allergic reactions, and inflammation. The present study was conducted to evaluate the protective effect of eckmaxol, a phlorotannin isolated from

Cracking and erosion are critical factors that reduce the mechanical properties and stability of concrete structures and soil, respectively. They are recognized worldwide as severe disasters causing the collapse of many structures including stone heritage and dams, and landslides. Therefore, it is essential to propose effective and environment-friendly management methods to prevent them. Carbonatogenesis has recently received considerable attention as a reliable biological process for remediating cracks in calcareous structures, stabilizing loose soils, and sequestering CO

A Gram-staining-negative, aerobic, light brown pigment bacterium, designated strain CE80T was isolated from marine sponge Callyspongia elegans in Jeju Island, Republic of Korea. Strain CE80T grew optimally at 25°C, in the range of pH 5.0-11.0 (optimum 7.0-8.0), and with 1.0-5.0% NaCl (optimum 1-3% (w/v)). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain CE80T belonged to the genus Labrenzia and was closely related to L. suaedae YC6927T (98.3%), L. alexandrii DFL-11T (96.6%), L. aggregata IAM 12614T (96.6%) L. marina mano18T (96.5%) and L. alba CECT 5094T (96.2%). The major fatty acids of strain CE80T were C18:1 ω7c, and summed feature. The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamin, one unidentified aminolipid, one phospholipid and four unidentified lipids. The DNA G+C content of strain CE80T was 55.9 mol%. The major respiratory quinone was Q-10. DNA-DNA relatedness between strain CE80T and L. suaedae YC6927T was 56.1±2.8%. On the basis of physiological and biochemical characterization and phylogenetic and chemotaxonomic analysis, strain CE80T represents a novel species of the Labrenzia, for which the name Labrenzia callyspongiae sp. nov., is proposed. The type strain is CE80T (=KCTC 42849T =JCM 31309T).

A bacterial strain, labeled UR11T, was isolated from green alga Ulva pertusa collected from Jeju Island, Korea. UR11T was identified as a gram-negative, rod-shaped, motile by gliding and aerobic bacterial strain with yellow colonies on R2A plates. The strain UR11T grew over at a temperature range of 10°C to 30°C (optimally at 25°C), a pH range of 6.0-11 (optimally at pH 7.0) and a Nacl range of 0.5-5% Nacl (w/v). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain UR11T was a member of the genus Flavobacterium. Strain UR11T shared close similarity with F. jejuensis EC11T (98.0%) F. jumunjinense HME7102T (96.1%), F. haoranii LQY-7T (95.3%), F. dongtanense LW30T (95.1%), and F. ahnfeltiae 10Alg 130T (94.9%). The major fatty acids (>5%) were iso-C15:0 (33.9%), iso-C15:1 G (12.4%), iso-C17:0 3-OH (9.0%), iso- C16:0 (7.0%) and iso-C15:0 3-OH (6.3%). The major polar lipids were phosphatidylethanolamine, seven unknown aminolipids, two unknown aminopolarlipids and two unknown lipids. DNADNA hybridization value was 58% at strain UR11T with F. jejuensis EC11T . Based on phenotypic, chemotaxonomic and phylogenetic evidence, strain UR11T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium jocheonensis sp. nov. is proposed. The type strain is Flavobacterium jocheonensis is UR11T (=KCTC 52377T =JCM 31512T.

This manuscript discusses about size and shape-selective preparation of Zinc oxide nanomaterials (ZnO NMs) using simple and handy soft jet plasma source. The soft jet was operated in an atmospheric pressure with low energy consumption and utilized ambient air as a feed gas. The plasma synthesized ZnO NMs exhibited variety of surface morphology such as nanorods, nanosheets and micro-size flowers as compared to the conventional low temperature synthesized powders which showed spherical shape. The precursor concentrations and kinds of precursors were strongly influenced into the NMs morphology and possible growth mechanism in the presence of plasma has been proposed. It is suggested here that the soft jet plasma is an effective tool to control size and shape of ZnO NMs especially when the preparations demanding low temperature, surfactant free and cost-effective synthesis. The prepared ZnO NMs were tested for its shape dependent anti-bacterial property against Gram-positive bacteria namely S. iniae and S. parauberis and two Gram-negative bacteria such as E. coli and V. anguillarum . The flower-like ZnO NMs showed more powerful anti-bacterial activity than rod or sheet-like ZnO structures.

Heavy metals such as gallium (Ga) cause serious physiological damage to exposed organisms, mostly of aquatic species. Ga one of the inter-metallic, transition elements increasingly being used in making high-speed semiconductors, such as Ga arsenide. The purposes of this study were to investigate the effects of Ga on acute toxicity, serum biochemical changes, and erythrocyte morphological changes in the blood stream of goldfish (Carassius auratus). Median lethal concentrations were determined in acute tests. The 96 hr LC 50 value was 9.15 mg/ml. Goldfish were exposed to different Ga concentrations (2.0, 4.0, and 8.0 mg/ml) for 30 days to assess its toxic effects. The results indicate that the measured serum biochemistry parameters (including glucose, blood urea nitrogen, creatinine, cholesterol, and triglyceride) of the Ga-exposed fish groups differed significantly from the untreated fish group. In addition, a change in the erythrocytes’ morphology at a high concentration (8.0 mg/ml) of Ga exposure shows respiratory problems. Our results suggest that 2.0 mg/ml is proposed as a biologically safe concentration that can be used for establishing tentative water quality criteria concerning the same-size goldfish.

Industrial advancements have resulted in food culture development, followed by increased seafood consumption and large-scale seafood farming, which has been accompanied by an increased preva-lence of fish disease. The antibiotic oxytetracycline (OTC) is commonly used to prevent and treat bac-terial diseases in fish. However, overuse of OTC had led to negative aspects. In view of this, we con-ducted a research with regard to aspects of remnants on olive flounder skin, liver, and muscle through dipping treatment and oral feeding of OTC and analyzed the results with bioassay and HPLC quantitative analyses. The dipping treatment was carried out once with 25 g/ton/hr of OTC, and the oral treatment with 62.5 mg/kg body weight/7 days. The results underwent a bioassay analysis. The dipping group reacted only on the skin right after dipping, while the oral feeding group responded on the skin for 77 days after feeding and on the muscle for 14 days. In the dipping group, the HPLC quantitative analysis revealed remnants in the skin on the 37th day and on the 13th day in the liver group. No remnants were found in the muscle, even immediately after dipping. In the oral feeding group, there was a high concentration (1.07 mg/kg) of remnant in the skin, even on the 77th day. 0.56 mg/kg in the liver, even a small amount, and no remnant in the muscle on the 42nd day. To sum up, the results suggest that it will not be harmful to our body to observe the OTC withdrawal period of 40 days with the muscle because OTC will hardly remain on it. When using olive flounder for sashimi, the skin and liver should not be used for broth, as the quantity of OTC residue is several times higher than that found in muscle. As previous studies reported that the concentration of rem-nants gradually decreased with heating, so it was likely to lessen, depending on the cooking temperature.

Streptococcus iniae is a causative agent of hemorrhagic septicemia in olive flounder, Paralichthys olivaceus, in Korea, resulting in serious economic losses. As a preventive measure, M VAC INIAE (Mastuken, Japan) was prepared from the S. iniae F2K strain and tested against the SI-36 strain prevalent on flounder fish farms on Jeju Island, Korea. F2K had a serotype of 38 (–) and SI-36 38 (+). The vaccine recognized both serotypes. It showed a very high effective immune response against S. iniae; the challenge test using the S. iniae SI-36 strain resulted in a relative percent survival (RPS) of 85.7-87.0% 2 weeks after vaccination and 71.0-80.0% 6 months after vaccination. Field vaccination and clinical challenge tests were performed at local Jeju aquafarms with S. iniae SI-36. These showed significantly reduced cumulative mortality when compared to the control group with RPS rates that ranged between 71-80%. Hence, the present study suggests that this vaccine showed a significant immune response against S. iniae and could be applied in commercial aquafarms as a therapeutic agent against â-hemolytic streptococcosis in cultured P. olivaceus. CORRESPONDING AUTHOR: Y.U. Jeong, D. Subramanian, D.H. Kim, S.H. Park, M.S. Heo [+] Marine Pathogenic Microbes Lab, Department of Aquatic Biomedical Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju 690-756, South Korea e-mail: msheo@jejunu.ac.kr Y.-H. Jang Jeju Special Self-Governing Province Ocean and Fisheries Research Institute, Pyoseon-myeon, Segwipo-si, Jeju 697-914, South Korea K. Park Department of Aquatic Life Medicine, Kunsan National University, Kunsan 573-701, South Korea Y.D. Lee Marine and Environmental Institute, Jeju National University, Jeju 690-814, South Korea Y.U. Jeong and D. Subramanian contributed equally to this work.

Eleven protease-producing bacteria were isolated from the organisms’ external shells and the inorganic materials collected from intertidal zone of Jeju Island, Republic of Korea. The samples were diluted serially, inoculated on Zobell agar plates with 1% skim milk and incubated at 20°C. Clear zone forming colonies were selected as protease-producing bacteria and each strain was identified based on the phylogenetic analysis with their 16S rDNA sequences. Strains JJM125, JJM129, YG47 and YG49 belong to the marine bacterial genus Pseudoalteromonas; strain JJM122 belong to the genus Microbulbifer; strains YG51, YG52, YG62 and YG63 belong to the genus Vibrio; and strain YG65 belong to genus Bacillus. Hence, the present study suggests that these protease producing bacteria could be further used to develop new varieties of protease with various biotechnological applications.

A bacterial strain, designated EC11(T) was isolated from brown alga Ecklonia cava collected from Jeju Island, Korea. EC11(T) was identified as a Gram-negative, rod-shaped and yellow-pigmented bacterial strain. The strain EC11(T) grew over a temperature range of 10 °C to 30 °C (optimally at 25 °C), and a pH range of 6.0-10.5 (optimally at pH 7.5). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EC11(T) belongs to the genus Flavobacterium. Strain EC11(T) shared close similarity with Flavobacterium jumunjinense HME7102(T) (96.4%), Flavobacterium dongtanense LW30(T) (95.8%), Flavobacterium haoranii LQY-7(T) (95.3%), and Flavobacterium urocaniciphilum (95.1%). The major fatty acids (> 5%) were iso-C17:0 3-OH (22.4%), iso-C15:0 3-OH (19.0%), C15:0 (12.4%), summed feature 3 (comprising C16:1 ω7c/ C16:1 ω6c; 9.78%), iso-C15:1 G (9.6%), and iso-C16:0 3-OH (9.0%). The DNA G+C content was 28.1 mol% and the strain contained MK-6 as the predominant menaquinone. The major polar lipids were phosphatidylethanolamine, two unknown aminolipids and three unknown polar lipids. Based on phenotypic, chemotaxonomic and phylogenetic analysis, strain EC11T represents a novel species of the Flavobacterium genus, for which the name Flavobacterium jejuensis sp. nov. is proposed. The type strain of F. jejuensis is EC11(T) (=KCTC 42149(T) = JCM 30735(T)).

The present study evaluates the growth performance of wild and cultured strains of giant freshwater prawn (GFP) in 2 × 2 diallel crosses as an initiation to improve its aquaculture production. A wild population was sampled from Vembanad Lake (Kerala) and from the farm-reared strain domesticated and cultured for several generations from grow-out ponds (Tamilnadu). From the four cross-combinations juveniles were produced using single pair mating. A total of 3,840 prawns (1,528 males and 2,312 females) were harvested after 15 weeks of grow-out hapas culture. The fixed effect included in the model was sex (male or female) and the age at harvest fitted as linear covariates. Total length, total weight, and carapace length of each prawn were recorded at the harvest. Additive genetic estimates of the crosses showed that the wild stock had higher length and weight (12.17% and 22.33%), while the cultured stock had lower length and weight (−18.26% and −22.98%). However, the average heterosis in the crossbred...

Present study deals with the preparation of differently shaped copper oxide nanomaterials (CuO NMs) using simple, reproducible wet chemical and hydrothermal approach. The influence of polyethylene glycol on the morphological control was also studied. The prepared nanomaterials exhibited new types of surface morphologies such as rice grain-like, needle-like and plate-like. The Fourier transform infrared spectra and the X-ray photoelectron spectra of all the samples showed identical chemical functional groups and did not exhibit the presence of any impurities. They exhibited good stability in the aqueous environment with the zeta potential values between −25 and −27 mV. The shape dependent anti-bacterial property of the prepared CuO NMs was evaluated against two Gram-positive bacteria namely Streptococcus iniae and Streptococcus parauberis and two Gram-negative bacteria such as Escherichia coli and Vibrio anguillarum . Among three CuO NMs, the plate-like CuO NMs displayed more powerful antibacterial activity than grain or needle shaped CuO NMs.

This study evaluated the pathophysiological, biochemical, and immunological status of olive flounder (Paralichthys olivaceus) infected with the myxosporean parasite Kudoa septempunctata. Flounder fish collected from Kudoa-infected and uninfected farms were confirmed by microscopic and TaqMan probe-based quantitative PCR screening. Morphological, biochemical, histological, and immune gene expression analyses were performed on uninfected and infected hosts to assess the effect of K. septempunctata. Histological studies confirmed the presence of Kudoa myxospores in the trunk muscles of infected flounder fish. Serum biochemical parameters, including the levels of myeloperoxidase activity, superoxide dismutase activity, alanine aminotransferase, alkaline phosphatase, amylase, bilirubin, total protein, cholesterol, calcium, potassium, sodium, phosphorus, glucose, and galactose, were found to exhibit no significant variations (p > 0.05) between uninfected and infected flounder fish. However, immune-related genes such as Mx, lysozyme, signal transducer and activator of transcription 1, interferon-γ, interferon regulatory factor, and tumour necrosis factor showed significantly elevated expression (p

Actinomycetes strain S-1 containing antioxidant components was isolated from the coast of Jeju Island in Korea and identified to the genus level using 16S ribosomal DNA sequencing and cellular fatty acids analysis as Nocardiopsis sp. (S-1). S-1 cultured supernatants showed high DPPH radical scavenging activities and hydroxyl radical scavenging abilities of up to 53%. Nutritional and culture conditions for antioxidant production were optimized. Galactose and yeast extracts were the best carbon and nitrogen sources for growth and antioxidant production. An initial medium acidity of pH 7.6, an incubation temperature of 25°C, a sodium chloride concentration of 2.5%, and an incubation time of 8 days were optimal. The DPPH scavenging activity of culture supernatants was 88% under optimum conditions. The S-1 strain can be useful for the functional fermented food and medicine industries.

In this study, we isolate and identify bacteria from seawater collected from Jeju coast, to evaluate the antimicrobial activity against the fish pathogenic bacteria. 14 bacterial strains were isolated and identified using physiological, biochemical and molecular tools. Antibacterial activity of all the 14 isolates were screened against four major fish pathogens namely, two Gram-positive: Streptococcus iniae, Streptococcus parauberis and two Gram-negative: Vibrio anguillarum, Edwardsiella tarda. Results revealed that among the 14 isolates, MK-11 was found to have antibacterial activity against S. iniae, S. parauberis, V. anguillarum Particularly, S. iniae was susceptibility with the MIC value of 250 μg/ml. The biochemical and physio-chemical results reveal that MK-11 had the sugar-alcohol disassemble ability of the D-sorbitol and D-mannitol. Also the utilization of the yeast extract, sorbitol and di-potassium phosphate were noted to be high. The optimum culture condition such as pH and temperature was recorded as pH 6.0, 25°C and along with 1% NaCl which differs from the previous reports particularly in nutrient resolutions. As results of the analysis of 16S rDNA sequences, MK-11 show the high similarity with Paenibacillus polymyxa, P. jamilae, P. brasilensis 99.78%, 99.43%, 99.39%, repectively. Hence, in the present study, the isolated Paemibacillus sp. MK-11 from Jeju seawater possesses the antibacterial activity against fish pathogens and it could be used as a new antibiotic agents against the gram positive fish pathogens.

Olive flounder (Paralichthys olivaceus) is an important aquaculture fish species in Jeju Island, South Korea. Due to the intensification of flounder fish farming, huge amounts of chemical antibiotics are used against several fish diseases. This has many harmful side effects on fish, as well as human consumers. Hence, an alternative to chemical antibiotic agents is needed for disease control. In this study, three strains of rhizobacteria (BRH433-2, TRH415-2, and THJ609-3) were isolated from the rhizosphere of plants. Assays of their antibacterial activity against fish pathogens, such as S. iniae, S. parauberis, V. anguillarum, and E. tarda, were performed with untreated broth culture (without cell separation), supernatant, and precipitated pellets separated by centrifugation. Among these, the cell suspension prepared from the precipitated pellet showed significant antimicrobial activity when compared with that of the untreated broth culture and centrifugal supernatant. These results indicate that the three isolated rhizobacterial strains exhibit antibacterial activity. Analysis of the 16S rDNA sequences of the BRH-433-2, THJ609-3, and TRH415-2 strains showed the highest similarity to Burkholderia gladioli (99.5%), Pseudomonas baetica (97.7%), and P. koreensis and P. baetica (98.4%), respectively. We suggest that the strains hold promise in disease management of fish.

The aim of this study was to investigate the bacterial community inhabited in Callyspongia elegans. Marine bacteria were isolated from the marine sponge C. elegans using marine agar. The resulting 112 isolated pure cultures were then used for further study. They were characterized by determining morphological characteristics through Gram`s staining and morphological observation. The colony pigments of bacterial isolates were characterized as yellow, brown, ivory, and white. Thirty-seven strains were found to be Gram-positive and 75 strains were Gram-negative. Seventy-nine strains were coccus-shaped, while 16 strains were rod-shaped. On the basis of the results of the comparative analyses of 16S rDNA gene sequences, the 112 isolated bacteria were divided into 5 major groups: Alphaproteobacteria (39%), Gammaproteobacteria (22%), Actinobacteria (14%), Fimicutes (9%), and Bacteroidetes (6%). It is strongly suggested that fifteen isolates are candidates for a new genera or species, based on the analyses of 16S rDNA gene sequences.

Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

The cell-derived extracellular matrix (ECM) membrane is a bio-absorbable membrane composed of aseptic ECM obtained from swine knee chondrocyte. The purpose of our study was to evaluate the inhibitory effect of the cell-derived ECM membrane in postoperative adhesion following strabismus surgery on rabbit eyes. Twenty four adult male New Zealand albino rabbits were used. Each rabbit has undergone 3 mm resection of the superior rectus muscle in both eyes. The cell-derived ECM membrane was applied between the conjunctiva and superior rectus muscle only to right eye in each rabbit. The eyes were divided into a surgery-ECM group and a surgery-no ECM group. The rabbits were sacrificed at 2, 4, and 6 weeks after the surgery. Each time, 16 eyes of 8 rabbits were enucleated to observe the adhesion in muscle and adjacent tissue grossly by blunt dissection. Then, histopathological sections were analyzed for inflammation and fibrosis by staining with hematoxylin-eosin and Masson trichrome, respectively. Inflammation and fibrosis were graded on a scale of 0 to 4. A researcher was blind to the experiments performed all the examinations. Comparing with the surgery-no ECM group, there was no significant difference with regard to inflammation at the area of superior rectus muscle resection at 2, 4, and 6 postoperative weeks (p = 0.52, p = 0.55, p = 0.82). However, a significant reduction in the degree of adhesion (p = 0.01, p = 0.03, p = 0.04) as well as decreased fibrosis (p = 0.04, p = 0.03, p = 0.02) between muscle and conjunctiva after strabismus surgery was observed in all surgery-ECM groups. The current results demonstrated that the cell-derived ECM membrane could inhibit the formation of postoperative adhesion and fibrosis between conjunctiva and muscle. The authors concluded that the use of ECM membrane could reduce postoperative adhesion around surgical area after strabismus surgery in rabbits.

The outbreak of haemorrhagic speticemia and dermal lesions in cultured Rock sea breem in Jeju Island are frequently associated with keratitis. Prolonged infection leads to the symptoms accompanied by necrosis of fin and skin and the fishes died. Present study aims to study the histopathological changes of diseased fish muscle tissue. The tissue at the initial stage of infection showed normal muscular texture, however in the late stages of infection, muscular lesion had been expanded and hence, necrosis had progressed deeply into muscle tissue. Mild tissue in caudal fin was found with more necrosis, and gill tissues were also collapsed. In Kidney, renal tubules were teared along with tissue destruction (Espada, J et al, 1993). Hence in the present study, we examined the histopathological variation of the infection fishes, and this basic data would be used for future research of in haemorrhagic specticemia and dermal diseases.

The strains were added to the feed in the concentration of , , and CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish`s immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

Spherical and sheet-like Ruthenium oxide (RuO2) nanomaterials were prepared by wet chemical approach. A new kind of sheet-like morphological features in RuO2 (with length and width >1 μm and 250 nm, respectively) was obtained by using polyethylene glycol (PEG) surfactant. The physical and chemical characterization for the synthesized RuO2 nanomaterials were carried out with the help of field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. The prepared nanomaterials were assessed for the shape dependent anti-microbial activities against fish pathogenic bacteria such as Vibrio anguillarum, Edwardsiella tarda, Streptococcus iniae and Streptococcus parauberis. Energy dispersive X-ray spectroscopy (EDS mapping) displayed the adsorption of nanomaterials on the bacterial surface. Both types of nanomaterials exhibited higher antibacterial activity against Gram-negative bacteria and in particular, sheet-like RuO2 nanomaterials showed more inhibitive effect than spherical types.

A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM 15406 T and Oceanisphera donghaensis KCTC 12522 T , respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to 37℃. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in C 16:1 ω7c and/or iso-C 15:0 2 -OH, C 16:0 , and C 18:1 ω7c fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

Two consecutive studies were conducted to evaluate the dietary supplementation of citrus by-products (CB) fermented with probiotic bacteria on growth performance, feed utilization, innate immune responses and disease resistance of juvenile olive flounder. In Experiment I, five diets were formulated to contain 0% (control) or 3% four different CB fermented with Bacillus subtilis (BS), Enterococcus faecium (EF), Lactobacillus rhamnosus (LR) and L. plantarum (LP) (designated as CON, CBF-BS, CBF-EF, CBF-LR and CBF-LP, respectively). During 10 weeks of a feeding trial, growth performance and feed efficiency were not significantly different among all the fish groups. However, fish fed CBF containing diets had significantly higher survivals than the CON group. Disease resistance of fish against Edwardsiella tarda was increased by the fermentation of CB. In Experiment II, we chose the BS as a promising probiotic and formulated five diets to contain 0%, 2%, 4%, 6% and 8% CBF-BS. Growth performance was not significantly affected by the CBF-BS supplementation during 6 weeks of a feeding trial. Innate immunity of fish was significantly enhanced by CBF-BS supplementation. Myeloperoxidase and lysozyme activities were increased in a dose-dependent manner by dietary CBF-BS inclusions. In a consecutive challenge test against E. tarda, an increased disease resistance was found by CBF-BS supplementation. These studies indicate that the fermentation process of CB with probiotic has beneficial effects on innate immunity and thereby increases disease resistance of olive flounder against E. tarda. Bacillus subtilis can be used as a promising probiotic microbe for by-product fermentation in fish feeds.

Respiratory burst, lysozyme, phagocytic activities, and immune gene expression levels were assessed in olive flounder (Paralichthys olivaceus, 10.52±2.5 g) fed with 0%, 0.025%, 0.05%, 0.1%, 0.2%, and 0.4% Lonicera japonica leaf powder (LjLP) for four weeks. All flounder fish fed LjLP showed significantly increased respiratory burst, lysozyme and phagocytic activity. Total immunoglobulin levels in LjLP diet fed fish groups were higher than in fish fed the basal diet (BD). In flounder fed LjLP, immune gene expression and antioxidant activity were significantly enhanced after 4 weeks of culture. One week following a challenge with Vibrio anguillarum, fish fed with different concentrations of LjLP showed decreased cumulative mortality and enhanced immunity all compared to the BD diet group. Our findings have shown that Lonicera japonica leaf powder enhanced immune response and resistance to V. anguillarum infection in olive flounder.

We investigate the efficacy of poly D,L-lactide-co-glycolic acid (PLGA)-encapsulated vaccine on innate and adaptive immune response in kelp grouper (Epinephelus bruneus) against Uronema marinum at weeks 1, 2, and 4. The respiratory burst (RB) activity, complement activity, and α2-macroglobulin were significantly enhanced in fish immunization with vaccine on week 4 whereas vaccine and PLGA-encapsulated vaccine from weeks 1 to 4. The serum lysozyme activity, antiprotease activity, and antibody level were significantly enhanced in fish immunized with vaccine and PLGA-encapsulated vaccine on weeks 2 and 4. The cumulative mortality was low in PLGA-encapsulated vaccine with 20% whereas high in PLGA and vaccine with 40% and 30%. The results from the present study suggest that PLGA-encapsulated vaccine is useful for further design of immunoprophylatic nano formulation against scuticociliatosis.

The efficacy of poly d , l -lactide-co-glycolic acid (PLGA)–liposome (L) encapsulated oligodeoxynucleotides with unmethylated deoxycytidyl-deoxyguanosine motifs (CpG-ODNs) on innate and adaptive immune response and disease resistance in kelp grouper (Epinephelus bruneus) against Vibrio alginolyticus at weeks 1, 2, and 4 is reported. The superoxide dismutase (SOD), respiratory burst, and lysozyme activities significantly increased in E. bruneus when immunized with ODN, PLGA + ODN, L + ODN, and PLGA + L + ODN on weeks 2 and 4. The serum complement activity was significantly enhanced with L + ODN and PLGA + L + ODN on week 1 while it increased with PLGA + ODN, L + ODN, and PLGA + L + ODN on weeks 2 and 4. The antibody titre consistently was increased with PLGA or L encapsulated with ODN (PLGA + ODN, L + ODN, and PLGA + L + ODN) from weeks 1 to 4. The cumulative mortality was 20% each in PLGA + ODN administered groups and 15% each in ODN, L + ODN, and PLGA + L + ODN groups during a period of 30 days. The present study suggests that PLGA–liposome encapsulated ODN has the potential to modulate the immune system and can serve as a useful tool for further design of immunoprophylatic nano drug formulations against bacterial diseases.

The aim of the present study was to determine the changes in the hematological and immunological response in kelp grouper Epinephelus bruneus against Vibrio harveyi infection after fed with 0%, 0.1%, 1.0%, and 2.0% Chaga mushroom, Inonotus obliquus extract enriched diet. All the doses significantly influenced the weight gain (WG) from weeks 1 to 4 when compared to the control. Other determined growth performance did not have any significant change at any time, except protein efficiency ratio in 2.0% diet as compared to control. The white blood cell and red blood cell levels, hemoglobin, haematocrite, lymphocytes, and monocytes significantly increased in infected fish fed with all diets. Total protein contents, albumin, and globulin level significantly increased with 1.0% and 2.0% than those of 0.1% diet from weeks 1 to 4. The infected fish fed with 1.0% and 2.0% diet significantly enhanced phagocytic activity from weeks 1 to 4. The respiratory burst and anti-protease activities significantly enhanced in infected fish fed with any doses of enriched diet on weeks 2 and 4. The lysozyme activity and serum bactericidal activity significantly increased in all doses of enriched diet from weeks 1 to 4. The cumulative mortality was low in 1.0% and 2.0% diets with 20% and 15% than those of 0.1% diet. This study suggest that 1.0% and 2.0% doses of diet significantly influence on growth performance and hematology, and could positively enhance innate immune system that effectively promote the health status of kelp grouper against V. harveyi infection.

The effect of different solvent-based (methanol, ethanol, and aqueous) extracts of Liriope platyphylla at 0, 0.1, 1.0, and 2.0 mg doses on hematology and innate immune response in olive flounder, Paralichthys olivaceus, against Flexibacter maritimus is reported. The red blood cells and white blood cells significantly increased with ethanol and aqueous extracts with any dose against F. maritimus while the hematocrit level significantly increased with methanol or ethanol extracts. However, the hemoglobin content significantly increased irrespective of the doses and type of extracts, whereas the mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration significantly decreased. The lymphocytes and monocytes significantly increased in all doses and extracts, whereas neutrophils increased in ethanol or aqueous extracts. However, the thrombocytes level neither the doses nor extracts did not significant changes to the control against pathogen. Total protein and globulin levels significantly increased with all the doses of ethanol extract while does not significant change in methanol extract. The phagocytic activity and complement activity were significantly enhanced with any dose of ethanol extract while with aqueous extract it was only at 1.0 and 2.0 mg doses. At all the doses of each extract significantly enhanced the leucocytes activity against F. maritimus. The results confirm that the ethanolic and aqueous extracts of L. platyphylla stimulate the immune response against F. maritimus in P. olivaceus.

The protective efficacy of liposomes-entrapped with whole cell bacterial vaccine (WCV) on immune response and protection against Vibrio harveyi infection in Epinephelus bruneus was investigated at weeks 1, 2, and 4. The cumulative mortality was 25% in WCV immunized fish whereas it was 35% and 40% with activated whole cell V. harveyi alone (WC) or liposome alone against pathogen; the mortality was 90% in non-immunization fish against pathogen. Total protein significantly increased in WCV and WC groups on weeks 2 and 4. The albumin, globulin, and albumin:globulin ratio significantly increased in WCV and WC groups on week 4. The antibody titres significantly increased in fish immunized with liposomes-entrapped WCV from weeks 1 to 4 where as in WC group only on week 4. The phagocytic activity significantly enhanced in liposomes-entrapped WCV group on weeks 1 and 2 and all other immunized groups on week 4. The complement activity was significantly enhanced in liposomes-entrapped WCV and WC immunized groups on weeks 2 and 4. However, the lysozyme activity significantly was enhanced in all immunization groups from weeks 1 to 4. These results suggest that fish immunized with liposomes-entrapped WCV and WC alone have the potential to modulate the immune system affording increased protection against V. harveyi infection in kelp grouper.

Philasterides dicentrarchi is a histophagous scuticociliate infecting Korean olive flounder farms in Jeju Island, South Korea, where it causes significant economic losses. However, the route of entry of these parasites in olive flounder is currently unknown. In the present study, we attempted experimental infection with different doses (concentrations) of P. dicentrarchi by intraperitoneal (IP) injection; maximum cumulative mortality of 95% was recorded on the administration of 2.1 × 108 ciliates/ml. In small size group (3 cm length, 24 ± 2 g wt) the ciliates were found in connective tissues of muscle, fins, and nervous tissue while in large size group (5 cm length, 47 ± 3 g wt), the ciliates were predominantly occurred with associated symptoms of liquefaction in the central nervous system. The major clinico-pathological manifestations were loss of scales, appearance of bleached spots that coalesced to form brownish patches, hemorrhagic, and severe dermal necrotic lesion. In addition there was degeneration of muscle fibers, necrosis of the hepatic parenchyma, and severe edema of the intestinal walls, extensive fouling, necrotic degeneration and hyperplasia in the gill bronchial epithelium. In this study, many ciliates were found in the blood vessel, brain, dorsal fins, muscle, kidney, and vertebra of infected flounder. The histopathological investigations are of significant importance in view of possible routes of entry into the host and doses of ciliates that trigger fast infection for potential utility of drugs as a strategy for the control of P. dicentrarchi in farmed olive flounder.