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Shao-Yang Hu
Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan

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Journal article
Published: 15 May 2020 in Sustainability
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Hepcidin antimicrobial peptides are difficult to produce in prokaryotic expression systems due to their complex structure and antimicrobial activity. Although synthetic hepcidin provides an alternative to solve this issue, its high cost limits its practical application in various industries. The present study used zebrafish eggs as bioreactors to produce convict cichlid (Amatitlania nigrofasciata) hepcidin (AN-hepc) using the oocyte-specific zona pellucida (zp3) promoter. The expression plasmid pT2-ZP3-AN-hepc-ZP3-EGFP, using EGFP as a reporter of AN-hepc expression, was designed to establish the transgenic line Tg(ZP3:AN-hepc:ZP3:EGFP) for the expression of AN-hepc. The AN-hepc peptide was produced successfully in fertilized eggs, as evidenced by RT-PCR and Western blotting. The AN-hepc-expressing eggs exhibited antimicrobial activity against a variety of aquatic pathogens and antibiotic-resistant pathogens, suggesting that the AN-hepc expressed in fish eggs was bioactive. The immunomodulatory effects of AN-hepc-expressing fertilized eggs on zebrafish innate immunity were evaluated by determining the expression of indicator genes after feeding with AN-hepc-expressing fertilized eggs for two months. Zebrafish supplementation with AN-hepc-expressing fertilized eggs significantly increased the expression of innate immunity-related genes, including IL-1β, IL-6, IL-15, TNF-α, NF-κb, complement C3b, lysozyme and TLR-4a. The zebrafish administered AN-hepc-expressing eggs exhibited higher cumulative survival than fish supplemented with wild-type and control eggs after infection with Aeromonas hydrophila and Streptococcus iniae. In conclusion, the present results showed that supplementation with AN-hepc-expressing fish eggs enhanced zebrafish innate immunity against pathogenic infections, suggesting that fertilized eggs containing AN-hepc have the potential to be developed as a food supplement for improving health status in aquaculture.

ACS Style

Chung-Chih Tseng; Tah-Wei Chu; Ridha Danata; Yenny Risjani; Hui-Tsu Shih; Shao-Yang Hu. Hepcidin-Expressing Fish Eggs as A Novel Food Supplement to Modulate Immunity against Pathogenic Infection in Zebrafish (Danio rerio). Sustainability 2020, 12, 1 .

AMA Style

Chung-Chih Tseng, Tah-Wei Chu, Ridha Danata, Yenny Risjani, Hui-Tsu Shih, Shao-Yang Hu. Hepcidin-Expressing Fish Eggs as A Novel Food Supplement to Modulate Immunity against Pathogenic Infection in Zebrafish (Danio rerio). Sustainability. 2020; 12 (10):1.

Chicago/Turabian Style

Chung-Chih Tseng; Tah-Wei Chu; Ridha Danata; Yenny Risjani; Hui-Tsu Shih; Shao-Yang Hu. 2020. "Hepcidin-Expressing Fish Eggs as A Novel Food Supplement to Modulate Immunity against Pathogenic Infection in Zebrafish (Danio rerio)." Sustainability 12, no. 10: 1.

Journal article
Published: 19 November 2019 in Sustainability
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Rice husk (RH) contains abundant silica such that RH silica (RHS) may be useful for possible industrial exploitation. Here, amorphous silica nanoparticles with multiple pore structures were acquired from RH by simple thermochemical processes. RHS antimicrobial activity and effects on zebrafish innate immunity against pathogen infections were evaluated. A toxicity assay showed that zebrafish exposed to an RHS dose lower than 200 μg/mL did not exhibit damage to zebrafish embryonic development or juvenile survival. RHS showed a wide spectrum of bacteriostatic activity against a variety of pathogens including antibiotic-resistant pathogens, implying its potential application as an antimicrobial agent in diverse industries. Fish exposed to 20 or 200 μg/mL RHS exhibited significantly increased mRNA expression of immune-related genes, including IL-1β, IL-6, IL-15, TNF-α, COX-2a, TLR-4a, lysozyme, and complement C3b. RHS-treated zebrafish exhibited a higher cumulative survival compared to that in control fish after infecting with Aeromonas hydrophila and Streptococcus iniae. The present results showed that a safe RHS dose enhanced innate immunity against infections without toxic effects in healthy fish, suggesting that RHS may be developed as an immunostimulant for improving health status in aquaculture.

ACS Style

Yong-Han Hong; Chung-Chih Tseng; Desy Setyoningrum; Zu-Po Yang; Maftuch; Shao-Yang Hu. Rice Husk Silica Enhances Innate Immune in Zebrafish (Danio rerio) and Improves Resistance to Aeromonas hydrophila and Streptococcus iniae Infection. Sustainability 2019, 11, 6504 .

AMA Style

Yong-Han Hong, Chung-Chih Tseng, Desy Setyoningrum, Zu-Po Yang, Maftuch, Shao-Yang Hu. Rice Husk Silica Enhances Innate Immune in Zebrafish (Danio rerio) and Improves Resistance to Aeromonas hydrophila and Streptococcus iniae Infection. Sustainability. 2019; 11 (22):6504.

Chicago/Turabian Style

Yong-Han Hong; Chung-Chih Tseng; Desy Setyoningrum; Zu-Po Yang; Maftuch; Shao-Yang Hu. 2019. "Rice Husk Silica Enhances Innate Immune in Zebrafish (Danio rerio) and Improves Resistance to Aeromonas hydrophila and Streptococcus iniae Infection." Sustainability 11, no. 22: 6504.

Journal article
Published: 01 October 2019 in Fish & Shellfish Immunology
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The use of probiotics as alternatives to antibiotics for disease control is a relatively eco-friendly approach in aquaculture; hence, studies isolating and assessing the benefit of potential probiotics to fish farming are common. The zebrafish is an excellent model system for validating beneficial functions of potential probiotics before their practical application in aquaculture. Here, a potentially probiotic Chromobacterium aquaticum was isolated from lake water samples and characterized by biochemical analysis and 16S rDNA sequencing. The probiotic produced extracellular enzymes (protease and xylanase) and a bacteriocin-like substance, which exhibited tolerance to extreme pH and high-temperature conditions and broad-spectrum bactericidal activity against diverse pathogens, including aquatic, foodborne, clinical and plant pathogens. The effects of C. aquaticum on zebrafish nutrient metabolism, growth performance and innate immunity were evaluated by measuring the expression of indicator genes after C. aquaticum feeding for 8 weeks. Fish administered the probiotic exhibited significantly increased hepatic mRNA expression of carbohydrate metabolism-related genes, including glucokinase (GK), hexokinase (HK), glucose-6-phosphatase (G6Pase), and pyruvate kinase (PK-L), and growth-related genes, including the growth hormone receptor (GHR) and insulin-like growth factor-1 (IGF-1). Innate immune-related genes (IL-1β, IL-6, TNF-α, IL-10, IL-21, NF-κb, lysozyme and complement C3b) were induced in fish with probiotic supplementation. Probiotic-treated fish exhibited a higher survival rate than control fish after challenge with Aeromonas hydrophila and Streptococcus iniae. Together, these data suggest that C. aquaticum, as a probiotic feed supplement, could enhance nutrient metabolism and growth performance and could modulate innate immunity against A. hydrophila and S. iniae in zebrafish.

ACS Style

Che-Chun Yi; Chun-Hung Liu; Kuo-Pin Chuang; Yi-Ting Chang; Shao-Yang Hu. A potential probiotic Chromobacterium aquaticum with bacteriocin-like activity enhances the expression of indicator genes associated with nutrient metabolism, growth performance and innate immunity against pathogen infections in zebrafish (Danio rerio). Fish & Shellfish Immunology 2019, 93, 124 -134.

AMA Style

Che-Chun Yi, Chun-Hung Liu, Kuo-Pin Chuang, Yi-Ting Chang, Shao-Yang Hu. A potential probiotic Chromobacterium aquaticum with bacteriocin-like activity enhances the expression of indicator genes associated with nutrient metabolism, growth performance and innate immunity against pathogen infections in zebrafish (Danio rerio). Fish & Shellfish Immunology. 2019; 93 ():124-134.

Chicago/Turabian Style

Che-Chun Yi; Chun-Hung Liu; Kuo-Pin Chuang; Yi-Ting Chang; Shao-Yang Hu. 2019. "A potential probiotic Chromobacterium aquaticum with bacteriocin-like activity enhances the expression of indicator genes associated with nutrient metabolism, growth performance and innate immunity against pathogen infections in zebrafish (Danio rerio)." Fish & Shellfish Immunology 93, no. : 124-134.

Journal article
Published: 01 September 2019 in Fish & Shellfish Immunology
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The application of probiotics as an eco-friendly alternative to antibiotics is an emerging strategy for sustainable aquaculture. In the present study, Rummeliibacillus stabekisii was isolated from the gut of Nile tilapia, and the effects of R. stabekisii on the growth, innate immunity, disease resistance, and gut microbiota of Nile tilapia (Oreochromis niloticus) were investigated. The results showed significantly increased weight gain (WG), feed conversion ratio (FCR), and feed efficiency (FE) in Nile tilapia fed R. stabekisii for 8 weeks compared to those in fish fed a control diet. Intestinal digestive enzymes such as protease, cellulase, and xylanase were also significantly increased in the R. stabekisii-administered groups. Enhanced cumulative survival was exhibited in fish fed R. stabekisii after challenge with Aeromonas hydrophila and Streptococcus iniae. Immune parameters such as the phagocytic activity, respiratory bursts, and superoxide dismutase of head kidney leukocytes; serum lysozyme activity; and expression of the cytokine genes interleukin-1β, tumor necrosis factor-α, transforming growth factor-β, and heat shock protein 70 were significantly elevated in fish fed R. stabekisii. Administration of R. stabekisii considerably increased the abundance of potential probiotics (Bacillus and Lactobacillus spp.) and reduced abundances of potential pathogenic bacteria (Streptococcus and Staphylococcus spp.) in fish intestines. The present study indicated that dietary supplementation with R. stabekisii improved the growth, immunity, disease resistance, and gut microflora of Nile tilapia. This research is the first study reporting that the genus Rummeliibacillus is a potential probiotic in animals, suggesting that R. stabekisii can be used as a feed additive to enhance the growth and health status in tilapia.

ACS Style

Herng Yih Tan; Sai-Wei Chen; Shao-Yang Hu. Improvements in the growth performance, immunity, disease resistance, and gut microbiota by the probiotic Rummeliibacillus stabekisii in Nile tilapia (Oreochromis niloticus). Fish & Shellfish Immunology 2019, 92, 265 -275.

AMA Style

Herng Yih Tan, Sai-Wei Chen, Shao-Yang Hu. Improvements in the growth performance, immunity, disease resistance, and gut microbiota by the probiotic Rummeliibacillus stabekisii in Nile tilapia (Oreochromis niloticus). Fish & Shellfish Immunology. 2019; 92 ():265-275.

Chicago/Turabian Style

Herng Yih Tan; Sai-Wei Chen; Shao-Yang Hu. 2019. "Improvements in the growth performance, immunity, disease resistance, and gut microbiota by the probiotic Rummeliibacillus stabekisii in Nile tilapia (Oreochromis niloticus)." Fish & Shellfish Immunology 92, no. : 265-275.

Journal article
Published: 09 July 2019 in Molecules
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The production of a bacteriocin-like substance with antimicrobial activity, named peocin, by the probiotic Paenibacillus ehimensis NPUST1 was previously reported by our laboratory. The present study aimed to identify peocin and increase the peocin yield by heterologous expression in Escherichia coli BL21(DE3). Peocin was identified as a DNA starvation/stationary phase protection protein, also called DNA-binding protein from starved cells (Dps), by gel overlay and LC-MS/MS analysis. For mass production of peocin, fed-batch cultivation of E. coli was performed using a pH-stat control system. Purification by simple nickel affinity chromatography and dialysis yielded 45.3 mg of purified peocin from a 20-mL fed-batch culture (49.3% recovery). The biological activity of the purified peocin was confirmed by determination of the MIC and MBC against diverse pathogens. Purified peocin exhibited antimicrobial activity against aquatic, food spoilage, clinical and antibiotic-resistant pathogens. In an in vivo challenge test, zebrafish treated with purified peocin exhibited significantly increased survival rates after A. hydrophila challenge. The present study is the first to show the antimicrobial activity of Dps and provides an efficient strategy for production of bioactive peocin, which will aid the development of peocin as a novel antimicrobial agent with potential applications in diverse industries.

ACS Style

Chung-Chih Tseng; Lini Murni; Tai-Wei Han; Diana Arfiati; Hui-Tsu Shih; Shao-Yang Hu; Han; Shih; Hu. Molecular Characterization and Heterologous Production of the Bacteriocin Peocin, a DNA Starvation/Stationary Phase Protection Protein, from Paenibacillus ehimensis NPUST1. Molecules 2019, 24, 2516 .

AMA Style

Chung-Chih Tseng, Lini Murni, Tai-Wei Han, Diana Arfiati, Hui-Tsu Shih, Shao-Yang Hu, Han, Shih, Hu. Molecular Characterization and Heterologous Production of the Bacteriocin Peocin, a DNA Starvation/Stationary Phase Protection Protein, from Paenibacillus ehimensis NPUST1. Molecules. 2019; 24 (13):2516.

Chicago/Turabian Style

Chung-Chih Tseng; Lini Murni; Tai-Wei Han; Diana Arfiati; Hui-Tsu Shih; Shao-Yang Hu; Han; Shih; Hu. 2019. "Molecular Characterization and Heterologous Production of the Bacteriocin Peocin, a DNA Starvation/Stationary Phase Protection Protein, from Paenibacillus ehimensis NPUST1." Molecules 24, no. 13: 2516.

Original article
Published: 04 January 2019 in Aquaculture Research
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The aims of this study were to unravel the intestinal microbiota of Litopenaeus vannamei after being fed a diet without (control) or with the synbiotic (SYN) for 60 days using next‐generation sequencing technology to see if changes in the intestinal microbiota were involved in the improved growth performance and health status of the shrimp. Next‐generation sequencing data showed that six phyla, 11 classes, 19 orders, 30 families, 58 genera and 73 species with taxonomic names assigned were detected. The majority of the operational taxonomic units (OTUs) was shared between the SYN and control shrimp and comprised 37 OTUs. However, intestinal biodiversity analyses revealed that SYN‐fed shrimp had a higher species richness, evenness and Shannon–Weaver index than did shrimp fed the control diet, but without reaching statistical significance. Interestingly, shrimp fed the SYN diet exhibited improved colonization of Lactobacillus plantarum and reduced prevalences of Vibrio harveyi and Photobacterium damselae in the intestines. These findings indicate that the SYN was able to modulate the intestinal bacterial community of shrimp and could be used to control vibriosis in shrimp.

ACS Style

Truong-Giang Huynh; Shao-Yang Hu; Chiu-Shia Chiu; Quoc-Phu Truong; Chun-Hung Liu. Bacterial population in intestines of white shrimp, Litopenaeus vannamei fed a synbiotic containing Lactobacillus plantarum and galactooligosaccharide. Aquaculture Research 2019, 50, 807 -817.

AMA Style

Truong-Giang Huynh, Shao-Yang Hu, Chiu-Shia Chiu, Quoc-Phu Truong, Chun-Hung Liu. Bacterial population in intestines of white shrimp, Litopenaeus vannamei fed a synbiotic containing Lactobacillus plantarum and galactooligosaccharide. Aquaculture Research. 2019; 50 (3):807-817.

Chicago/Turabian Style

Truong-Giang Huynh; Shao-Yang Hu; Chiu-Shia Chiu; Quoc-Phu Truong; Chun-Hung Liu. 2019. "Bacterial population in intestines of white shrimp, Litopenaeus vannamei fed a synbiotic containing Lactobacillus plantarum and galactooligosaccharide." Aquaculture Research 50, no. 3: 807-817.

Journal article
Published: 22 November 2018 in Fish & Shellfish Immunology
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Zebrafish (Danio rerio) are an excellent model for assessing the beneficial effects of probiotics before applying them in aquaculture. This study evaluated the effects on zebrafish of dietary supplementation with the probiotic Bacillus amyloliquefaciens R8, which heterologously expresses xylanase from rumen fungi. Nutrient metabolism, hepatic oxidative stress, and innate immunity against pathogen infections were investigated. Treated zebrafish received feed supplemented with B. amyloliquefaciens R8 for 30 days and then were compared to zebrafish that were fed a control diet. The treated fish showed significant increases in xylanase activity in the intestines. The livers of the treated fish showed increased mRNA expressions of glycolysis-related genes of hexokinase, glucokinase, glucose-6-phosphatase, and pyruvate kinase; and higher enzyme activities of 3-hydroxyacyl-coenzyme A dehydrogenase and citrate synthase which are associated with fatty acid β-oxidation and mitochondrial integrity. The livers of treated fish also showed decreased mRNA expressions of oxidative stress-related genes (SOD, Gpx, NOS2, and Hsp70) and an apoptotic gene (tp53), as well as increased expression of an anti-apoptotic gene (bcl-2). The probiotics-treated fish had increased expression of innate immune-related genes (IL-1β, IL-6, IL-21, TNF-α, and TLR-1, -3, and -4). Following challenge with Aeromonas hydrophila and Streptococcus agalactiae, treated fish showed increased a higher survival rate than control fish. Overall, results showed that the administration of xylanase-expressing B. amyloliquefaciens R8 can potentially improve nutrient metabolism and hepatic stress tolerance, and enhance immunity and disease resistance against A. hydrophila and S. agalactiae in zebrafish.

ACS Style

Yu-Sheng Lin; Febriyansyah Saputra; Yo-Chia Chen; Shao-Yang Hu. Dietary administration of Bacillus amyloliquefaciens R8 reduces hepatic oxidative stress and enhances nutrient metabolism and immunity against Aeromonas hydrophila and Streptococcus agalactiae in zebrafish (Danio rerio). Fish & Shellfish Immunology 2018, 86, 410 -419.

AMA Style

Yu-Sheng Lin, Febriyansyah Saputra, Yo-Chia Chen, Shao-Yang Hu. Dietary administration of Bacillus amyloliquefaciens R8 reduces hepatic oxidative stress and enhances nutrient metabolism and immunity against Aeromonas hydrophila and Streptococcus agalactiae in zebrafish (Danio rerio). Fish & Shellfish Immunology. 2018; 86 ():410-419.

Chicago/Turabian Style

Yu-Sheng Lin; Febriyansyah Saputra; Yo-Chia Chen; Shao-Yang Hu. 2018. "Dietary administration of Bacillus amyloliquefaciens R8 reduces hepatic oxidative stress and enhances nutrient metabolism and immunity against Aeromonas hydrophila and Streptococcus agalactiae in zebrafish (Danio rerio)." Fish & Shellfish Immunology 86, no. : 410-419.

Journal article
Published: 09 November 2018 in Fish & Shellfish Immunology
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The probiotic efficiencies of the mixed probiotics containing Lactobacillus pentosus BD6, Lac. fermentum LW2, Bacillus subtilis E20, and Saccharomyces cerevisiae P13 for shrimp growth and health status improvement were better than those when using single probiotics. The probiotic mixture at a level of 108 colony-forming units (cfu) (kg diet)−1 and the diets containing BD6 and E20 at 109 cfu (kg diet)−1 significantly improved the growth and health status of shrimp, whereas the diets containing P13 or LW2 did not significantly affect the growth of shrimp. No significant difference in the carcass composition was recorded among the control and treatments. After 56 days of feeding, shrimp fed the diet containing the probiotic mixture (107∼109 cfu (kg diet)−1) had higher survival after injection with the V. alginolyticus, but 109 cfu (kg diet)−1 of single probiotics (except for S. cerevisiae P13) had to be administered to improve shrimp survival. The better disease resistance of shrimp in groups fed the probiotic mixture might have been due to increased phenoloxidase activity, respiratory bursts, and lysozyme activity of hemocytes. Therefore, we considered that the probiotic mixture could adequately provide probiotic efficiency for white shrimp, and a diet containing 108 cfu (kg diet)−1 probiotic mixture is recommended.

ACS Style

Yu-Chu Wang; Shao-Yang Hu; Chiu-Shia Chiu; Chun-Hung Liu. Multiple-strain probiotics appear to be more effective in improving the growth performance and health status of white shrimp, Litopenaeus vannamei, than single probiotic strains. Fish & Shellfish Immunology 2018, 84, 1050 -1058.

AMA Style

Yu-Chu Wang, Shao-Yang Hu, Chiu-Shia Chiu, Chun-Hung Liu. Multiple-strain probiotics appear to be more effective in improving the growth performance and health status of white shrimp, Litopenaeus vannamei, than single probiotic strains. Fish & Shellfish Immunology. 2018; 84 ():1050-1058.

Chicago/Turabian Style

Yu-Chu Wang; Shao-Yang Hu; Chiu-Shia Chiu; Chun-Hung Liu. 2018. "Multiple-strain probiotics appear to be more effective in improving the growth performance and health status of white shrimp, Litopenaeus vannamei, than single probiotic strains." Fish & Shellfish Immunology 84, no. : 1050-1058.

Journal article
Published: 24 October 2018 in Fish & Shellfish Immunology
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Bacteria-induced diseases are a major cause of mortality in aquaculture. Probiotics have commonly been used to replace antibiotics for prophylactic biocontrol in aquaculture. In the present study, Paenibacillus ehimensis NPUST1 was isolated from a tilapia culture pond. This probiotic has bacteriocin-like activities against Aeromonas hydrophila and was characterized by biochemical analysis and 16S rDNA sequencing. The physiochemical properties of a crude extract of the bacteriocin-like substance revealed low pH and high thermal tolerance. The substance exhibited broad-spectrum antimicrobial activity against diverse aquatic pathogens, food spoilage, clinical pathogens, and plant pathogens. The effect of dietary supplementation with P. ehimensis NPUST1 was evaluated in regard to the growth of Nile tilapia (Oreochromis niloticus) and immunity against pathogenic infection. The results showed significantly increased weight gain (WG), feed conversion ratio (FCR), and feed efficiency (FE) in Nile tilapia fed P. ehimensis NPUST1 for 2 months compared with fish fed a control diet. When challenged with A. hydrophila and S. iniae, the fish fed P. ehimensis NPUST1 also exhibited a higher survival rate than fish fed the control diet. The immune parameters revealed that the P. ehimensis NPUST1-fed fish had significantly higher phagocytic activity, respiratory burst, and superoxide dismutase (SOD) of the head kidney leukocytes, as well as higher serum lysozyme activity and expression of cytokines TNF-α and IL-1β than the fish fed the control diet. These results indicate that dietary supplementation with P. ehimensis NPUST1 improved the growth performance, immunity, and disease resistance in Nile tilapia.

ACS Style

Sai-Wei Chen; Chun-Hung Liu; Shao-Yang Hu. Dietary administration of probiotic Paenibacillus ehimensis NPUST1 with bacteriocin-like activity improves growth performance and immunity against Aeromonas hydrophila and Streptococcus iniae in Nile tilapia (Oreochromis niloticus). Fish & Shellfish Immunology 2018, 84, 695 -703.

AMA Style

Sai-Wei Chen, Chun-Hung Liu, Shao-Yang Hu. Dietary administration of probiotic Paenibacillus ehimensis NPUST1 with bacteriocin-like activity improves growth performance and immunity against Aeromonas hydrophila and Streptococcus iniae in Nile tilapia (Oreochromis niloticus). Fish & Shellfish Immunology. 2018; 84 ():695-703.

Chicago/Turabian Style

Sai-Wei Chen; Chun-Hung Liu; Shao-Yang Hu. 2018. "Dietary administration of probiotic Paenibacillus ehimensis NPUST1 with bacteriocin-like activity improves growth performance and immunity against Aeromonas hydrophila and Streptococcus iniae in Nile tilapia (Oreochromis niloticus)." Fish & Shellfish Immunology 84, no. : 695-703.

Journal article
Published: 01 November 2016 in Fish & Shellfish Immunology
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Bacillus amyloliquefaciens has attracted attention as a probiotic in aquaculture due to its immunostimulatory activity against pathogenic infection. Xylanases are extensively used in animal feed to degrade plant ingredients, enhancing nutrient utilization and increasing the growth rate of various animals. In the present study, the effects of dietary supplementation with B. amyloliquefaciens and xylanase-expressing B. amyloliquefaciens R8 on the growth of Nile tilapia (Oreochromis niloticus) and immunity against Aeromonas hydrophila were evaluated. The results showed that the xylanase activity in the intestine, weight gain (WG), feed efficiency (FE) and condition factor (CF) of Nile tilapia fed B. amyloliquefaciens R8 for 2 months were significantly increased compared with those of the fish fed the control diet and B. amyloliquefaciens. Moreover, the mRNA expression of growth- and metabolism-related genes, such as insulin-like growth factor-1 (igf-1), glucokinase (GK), glucose-6-phosphate 1-dehydrogenase (G6PD), and glucose-6-phosphatase (G6Pase), was significantly induced in Nile tilapia fed administered B. amyloliquefaciens R8, and this group also exhibited a higher survival rate than the control fish following a challenge with A. hydrophila. The phagocytic activity and respiratory burst activity of head kidney leukocytes as well as the serum lysozyme activity of B. amyloliquefaciens R8-fed Nile tilapia were significantly higher than those of fish fed the control diet for 2 months. Superoxide dismutase (SOD) levels in the head kidney leukocytes of Nile tilapia fed B. amyloliquefaciens R8 differed from those of fish fed the control diet, but this was not significant. These results indicate that dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity and disease resistance against A. hydrophila in Nile tilapia.

ACS Style

Febriyansyah Saputra; Ya-Li Shiu; Yo-Chia Chen; Asthervina Widyastami Puspitasari; Ridha Handriany Danata; Chun-Hung Liu; Shao-Yang Hu. Dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity against Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus ). Fish & Shellfish Immunology 2016, 58, 397 -405.

AMA Style

Febriyansyah Saputra, Ya-Li Shiu, Yo-Chia Chen, Asthervina Widyastami Puspitasari, Ridha Handriany Danata, Chun-Hung Liu, Shao-Yang Hu. Dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity against Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus ). Fish & Shellfish Immunology. 2016; 58 ():397-405.

Chicago/Turabian Style

Febriyansyah Saputra; Ya-Li Shiu; Yo-Chia Chen; Asthervina Widyastami Puspitasari; Ridha Handriany Danata; Chun-Hung Liu; Shao-Yang Hu. 2016. "Dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity against Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus )." Fish & Shellfish Immunology 58, no. : 397-405.

Journal article
Published: 17 November 2015 in Journal of Biomedical Science
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Highly desaturated n-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are synthesized by desaturases and elongase. They exert hepatoprotective effects to prevent alcoholic fatty liver syndrome or cholestatic liver injury. However, it is unclear how n-3 PUFAs improve immune function in liver. Vibrio vulnificus, a gram-negative bacterial pathogen, causes high mortality of aquaculture fishes upon infection. Humans can become infected with V. vulnificus through open wounds or by eating raw seafood, and such infections may result in systemic septicemia. Moreover, patients with liver diseases are vulnerable to infection, and are more likely than healthy persons to present with liver inflammation following infection. This study quantified n-3 PUFAs and their anti-bacterial effects in Fadsd6 and Elvol5a transgenic zebrafish. Two transgenic zebrafish strains with strong liver specific expression of Fadsd6 and Elvol5a (driven by the zebrafish Fabp10 promoter) were established using the Tol2 system. Synthesis of n-3 PUFAs in these strains were increased by 2.5-fold as compared to wild type (Wt) fish. The survival rate in 24 h following challenge with V. vulnificus was 20 % in Wt, but 70 % in the transgenic strains. In addition, the bacteria counts in transgenic fish strains were significantly decreased. The expression levels of pro-inflammatory genes, such as TNF-α, IL-1β, and NF-κB, were suppressed between 9 and 12 h after challenge. This study confirms the anti-bacterial function of n-3 PUFAs in a transgenic zebrafish model. Fadsd6 and Elvol5a transgenic zebrafish are more resistant to V. vulnificus infection, and enhance survival by diminishing the attendant inflammatory response.

ACS Style

Chih-Lun Cheng; Shin-Jie Huang; Chih-Lu Wu; Hong-Yi Gong; Chuian-Fu Ken; Shao-Yang Hu; Jen-Leih Wu. Transgenic expression of omega-3 PUFA synthesis genes improves zebrafish survival during Vibrio vulnificus infection. Journal of Biomedical Science 2015, 22, 1 -13.

AMA Style

Chih-Lun Cheng, Shin-Jie Huang, Chih-Lu Wu, Hong-Yi Gong, Chuian-Fu Ken, Shao-Yang Hu, Jen-Leih Wu. Transgenic expression of omega-3 PUFA synthesis genes improves zebrafish survival during Vibrio vulnificus infection. Journal of Biomedical Science. 2015; 22 (1):1-13.

Chicago/Turabian Style

Chih-Lun Cheng; Shin-Jie Huang; Chih-Lu Wu; Hong-Yi Gong; Chuian-Fu Ken; Shao-Yang Hu; Jen-Leih Wu. 2015. "Transgenic expression of omega-3 PUFA synthesis genes improves zebrafish survival during Vibrio vulnificus infection." Journal of Biomedical Science 22, no. 1: 1-13.

Journal article
Published: 01 June 2015 in Protein Expression and Purification
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Growth hormone (GH) performs important roles in regulating somatic growth, reproduction, osmoregulation, metabolism and immunity in teleosts, and thus, it has attracted substantial attention in the field of aquaculture application. Herein, giant grouper GH (ggGH) cDNA was cloned into the pET28a vector and expressed in Shuffle® T7 Competent Escherichia coli. Recombinant N-terminal 6× His-tagged ggGH was produced mainly in insoluble inclusion bodies; the recombinant ggGH content reached 20% of total protein. For large-scale ggGH production, high-cell density E. coli culture was achieved via fed-batch culture with pH-stat. After 30h of cultivation, a cell concentration of 41.1g/l dry cell weight with over 95% plasmid stability was reached. Maximal ggGH production (4.0g/l; 22% total protein) was achieved via mid-log phase induction. Various centrifugal forces, buffer pHs and urea concentrations were optimized for isolation and solubilization of ggGH from inclusion bodies. Hydrophobic interactions and ionic interactions were the major forces in ggGH inclusion body formation. Complete ggGH inclusion body solubilization was obtained in PBS buffer at pH 12 containing 3M urea. Through a simple purification process including Ni-NTA affinity chromatography and refolding, 5.7mg of ggGH was obtained from 10ml of fed-batch culture (45% recovery). The sequence and secondary structure of the purified ggGH were confirmed by LC-MS/MS mass spectrometry and circular dichroism analysis. The cell proliferation-promoting activity was confirmed in HepG2, ZFL and GF-1 cells with the WST-1 colorimetric bioassay.

ACS Style

Wen-Jen Chung; Chi-Lung Huang; Hong-Yi Gong; Tsung-Yin Ou; Jue-Liang Hsu; Shao-Yang Hu. Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture. Protein Expression and Purification 2015, 110, 79 -88.

AMA Style

Wen-Jen Chung, Chi-Lung Huang, Hong-Yi Gong, Tsung-Yin Ou, Jue-Liang Hsu, Shao-Yang Hu. Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture. Protein Expression and Purification. 2015; 110 ():79-88.

Chicago/Turabian Style

Wen-Jen Chung; Chi-Lung Huang; Hong-Yi Gong; Tsung-Yin Ou; Jue-Liang Hsu; Shao-Yang Hu. 2015. "Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture." Protein Expression and Purification 110, no. : 79-88.

Journal article
Published: 01 January 2015 in Journal of Marine Science: Research & Development
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4-tert-octylphenol (4-t-OP), an environmental exogenous estrogen is produced by microbial degradation of alkylphenol polyethoxylates (APEOs). Although it is well known that 4-t-OP can cause the feminization of male, sterility and deficiency of gonad development of aquatic animals by disrupting the endocrine reproductive signaling, less is known about the effects of 4-t-OP on embryonic development. Moreover, the presence of 4-t-OP were detected in umbilical cord blood samples of newborns suggesting infants during development may expose to the risk of 4-t-OP contaminant, hence to investigate the effect of 4-t-OP on physiological function during embryonic development is necessary. In the present study, zebrafish embryos exposed to 4-t-OP were used to evaluate the toxicity of 4-t-OP. The 50% lethal dose (LD50) for wild type zebrafish embryos exposure to 4-t-OP for 3 days is approximately 1.0 μM, and a high ratio of cardiovascular defects were showed in survival embryos. To observe the cardiovascular defects more efficiently, Tg (fil-1: EGFP) zebrafish embryos was used in 4-t-OP exposure treatment. Following exposure Tg (fil-1: EGFP) zebrafish embryos to 4-t-OP at 1.0 μM for 4 days, a highly proportion of defects revealed in cardiovascular system, including pericardical edema, irregular shape or incomplete looping of ventricle and atrium, the absence of intersegmental vessel in the tail of notochord, unformed parachordal vessel and kinks in the caudal vein. The phenotype of cardiovascular defects was accompanied by reduced heart rate and impaired blood circulation. The mRNA expression levels of transcription factors, which are critical for zebrafish heart chamber formation and blood vessel development, were analyzed by RT-PCR. The results showed that the presence of 4-t-OP significantly induce expression level of ERα and ERβ2, and caused cardiovascular defects by suppressing transcription factor Nkx2.7, Hand2, Tbx2a, Tbx2b, Tbx5a, FGF1a, GATA-4, -5 and -6 in zebrafish. The present study suggests that 4-t-OP affects the cardiovascular development in zebrafish and elucidated that early life exposure to 4-t-OP potentially may take a risk of impaired cardiovascular function.

ACS Style

Saputra F; Chia-Hung Yen; Chi-Ying Hsieh; Tsung-Yin Ou; Risjani Y; Wee-Keat Cheah; Shao-Yang Hu. Toxicity Effects of the Environmental Hormone 4-Tert-Octylphenol in Zebrafish (Danio Rerio). Journal of Marine Science: Research & Development 2015, 6, 1 .

AMA Style

Saputra F, Chia-Hung Yen, Chi-Ying Hsieh, Tsung-Yin Ou, Risjani Y, Wee-Keat Cheah, Shao-Yang Hu. Toxicity Effects of the Environmental Hormone 4-Tert-Octylphenol in Zebrafish (Danio Rerio). Journal of Marine Science: Research & Development. 2015; 6 (1):1.

Chicago/Turabian Style

Saputra F; Chia-Hung Yen; Chi-Ying Hsieh; Tsung-Yin Ou; Risjani Y; Wee-Keat Cheah; Shao-Yang Hu. 2015. "Toxicity Effects of the Environmental Hormone 4-Tert-Octylphenol in Zebrafish (Danio Rerio)." Journal of Marine Science: Research & Development 6, no. 1: 1.

Journal article
Published: 04 October 2014 in Fish Physiology and Biochemistry
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The hepcidin gene is widely expressed in many fish species and functions as an antimicrobial peptide, suggesting that it plays an important role in the innate immune system of fish. In the present study, the hepcidin gene (AN-hepc) was cloned from the liver and its expression during an immune response was characterized. The results of quantitative PCR and RT-PCR showed that the AN-hepc transcript was most abundant in the liver. The expression of AN-hepc mRNA was significantly increased in the liver, stomach, heart, intestine, gill and muscle but was not significantly altered in the spleen, kidney, brain or skin after lipopolysaccharide challenge. The synthetic AN-hepc peptide showed a wide spectrum of antimicrobial activity in vitro toward gram-positive and gram-negative bacteria. In particular, this peptide demonstrated potent antimicrobial activity against the aquatic pathogens , , , and . The in vivo bacterial challenge results demonstrated that the synthetic AN-hepc peptide significantly improved the survival rate of - and -infected zebrafish. Taken together, these data indicate an important role for AN-hepc in the innate immunity of and suggest its potential application in aquaculture for increasing resistance to disease.

ACS Style

Jing-Ruei Chi; Long-Si Liao; Rong-Guang Wang; Chu-Sian Jhu; Jen-Leih Wu; Shao-Yang Hu. Molecular cloning and functional characterization of the hepcidin gene from the convict cichlid (Amatitlania nigrofasciata) and its expression pattern in response to lipopolysaccharide challenge. Fish Physiology and Biochemistry 2014, 41, 449 -461.

AMA Style

Jing-Ruei Chi, Long-Si Liao, Rong-Guang Wang, Chu-Sian Jhu, Jen-Leih Wu, Shao-Yang Hu. Molecular cloning and functional characterization of the hepcidin gene from the convict cichlid (Amatitlania nigrofasciata) and its expression pattern in response to lipopolysaccharide challenge. Fish Physiology and Biochemistry. 2014; 41 (2):449-461.

Chicago/Turabian Style

Jing-Ruei Chi; Long-Si Liao; Rong-Guang Wang; Chu-Sian Jhu; Jen-Leih Wu; Shao-Yang Hu. 2014. "Molecular cloning and functional characterization of the hepcidin gene from the convict cichlid (Amatitlania nigrofasciata) and its expression pattern in response to lipopolysaccharide challenge." Fish Physiology and Biochemistry 41, no. 2: 449-461.

Journal article
Published: 01 May 2014 in Fish & Shellfish Immunology
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Cinnamaldehyde, which is extracted from cinnamon, is a natural compound with activity against bacteria and a modulatory immune function. However, the antibacterial activity and immunostimulation of cinnamaldehyde in fish has not been well investigated due to the compound's poor water solubility. Thus, liposome-encapsulated cinnamaldehyde (LEC) was used to evaluate the effects of cinnamaldehyde on in vitro antibacterial activity against aquatic pathogens and in vivo immunity and protection parameters against Vibrio vulnificus and Streptococcus agalactiae. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as well as bactericidal agar plate assay results demonstrated the effective bacteriostatic and bactericidal potency of LEC against Aeromonas hydrophila, V. vulnificus, and S. agalactiae, as well as the antibiotic-resistant Vibrio parahaemolyticus and Vibrio alginolyticus. Bacteria challenge test results demonstrated that LEC significantly enhances the survival rate and inhibits bacterial growth in zebrafish infected with A. hydrophila, V. vulnificus, and S. agalactiae. A gene expression study using a real-time PCR showed that LEC immersion-treated zebrafish had increased endogenous interleukin (IL)-1β, IL-6, IL-15, IL-21, tumor necrosis factor (TNF)-α, and interferon (INF)-γ expression in vivo. After the zebrafish were infected with V. vulnificus or S. agalactiae, the LEC immersion treatment suppressed the expression of the inflammatory cytokines IL-1β, IL-6, IL-15, NF-κb, and TNF-α and induced IL-10 and C3b expression. These findings demonstrate that cinnamaldehyde exhibits antimicrobial activity against aquatic pathogens, even antibiotic-resistant bacterial strains and immune-stimulating effects to protect the host's defenses against pathogen infection in bacteria-infected zebrafish. These results suggest that LEC could be used as an antimicrobial agent and immunostimulant to protect bacteria-infected fish in aquaculture.

ACS Style

Elok Ning Faikoh; Yong-Han Hong; Shao-Yang Hu. Liposome-encapsulated cinnamaldehyde enhances zebrafish (Danio rerio) immunity and survival when challenged with Vibrio vulnificus and Streptococcus agalactiae. Fish & Shellfish Immunology 2014, 38, 15 -24.

AMA Style

Elok Ning Faikoh, Yong-Han Hong, Shao-Yang Hu. Liposome-encapsulated cinnamaldehyde enhances zebrafish (Danio rerio) immunity and survival when challenged with Vibrio vulnificus and Streptococcus agalactiae. Fish & Shellfish Immunology. 2014; 38 (1):15-24.

Chicago/Turabian Style

Elok Ning Faikoh; Yong-Han Hong; Shao-Yang Hu. 2014. "Liposome-encapsulated cinnamaldehyde enhances zebrafish (Danio rerio) immunity and survival when challenged with Vibrio vulnificus and Streptococcus agalactiae." Fish & Shellfish Immunology 38, no. 1: 15-24.

Journal article
Published: 10 August 2012 in FEBS Letters
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It is well known that growth hormone (GH)-induced IGF-1 signaling plays a dominant role in postnatal muscle growth. Our previous studies have identified a growth factor, progranulin (PGRN), that is co-induced with IGF-1 upon GH administration. This result prompted us to explore the function of PGRN and its association with IGF-1. In the present study, we demonstrated that, similar to IGF-1, PGRN can promote C2C12 myotube hypertrophy via the PI(3)K/Akt/mTOR pathway. Moreover, PGRN can rescue the muscle atrophy phenotypes in C2C12 myotube when IGF-1 signaling is blocked. This result shows that PGRN can substitute for IGF-1 signaling in the regulation of muscle growth. Our findings provide new insights into IGF-1-modulated complicated networks that regulate muscle growth

ACS Style

Shao-Yang Hu; Chen-Chen Tai; Yen-Hsing Li; Jen-Leih Wu. Progranulin compensates for blocked IGF-1 signaling to promote myotube hypertrophy in C2C12 myoblasts via the PI3K/Akt/mTOR pathway. FEBS Letters 2012, 586, 3485 -3492.

AMA Style

Shao-Yang Hu, Chen-Chen Tai, Yen-Hsing Li, Jen-Leih Wu. Progranulin compensates for blocked IGF-1 signaling to promote myotube hypertrophy in C2C12 myoblasts via the PI3K/Akt/mTOR pathway. FEBS Letters. 2012; 586 (19):3485-3492.

Chicago/Turabian Style

Shao-Yang Hu; Chen-Chen Tai; Yen-Hsing Li; Jen-Leih Wu. 2012. "Progranulin compensates for blocked IGF-1 signaling to promote myotube hypertrophy in C2C12 myoblasts via the PI3K/Akt/mTOR pathway." FEBS Letters 586, no. 19: 3485-3492.

Evaluation study
Published: 07 April 2010 in Transgenic Research
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Multiple advantages-including the short generation time, large numbers of fertilized eggs, low cost of cultivation and easy maintenance favor the use of fish as bioreactors for the production of pharmaceutical proteins. In the present study, zebrafish eggs were used as bioreactors to produce mature tilapia insulin-like growth factors (IGFs) proteins using the oocyte-specific zona pellucida (zp3) promoter. The chimeric expression plasmids, pT2-ZP-tIGFs-IRES-hrGFP, in which hrGFP was used as reporter of tilapia IGFs expression, were designed to established Tg (ZP:tIGFs:hrGFP) transgenic lines for the expression of tilapia IGF-1 and IGF-2. Recombinant tilapia IGF-1 and IGF-2 were expressed as soluble forms in cytoplasm of fertilized eggs. The content level of tilapia IGF-1 and IGF-2 were 6.5 and 5.0% of the soluble protein, respectively. Using a simple Ni–NTA affinity chromatography purification process, 0.58 and 0.49 mg of purified tilapia IGF-1 and IGF-2 were obtained, respectively, from 650 fertilized eggs. The biological activity of the purified tilapia IGF-1 and IGF-2 was confirmed via a colorimetric bioassay to monitor the growth stimulation of zebrafish embryonic cells (ZF4), tilapia ovary cells (TO-2) and human osteosarcoma epithelial cells (U2OS). These results demonstrate that the use of zebrafish eggs as bioreactors is a promising approach for the production of biological recombinant proteins.

ACS Style

Shao-Yang Hu; Chia-Hsuan Liao; Yi-Pei Lin; Yen-Hsing Li; Hong-Yi Gong; Gen-Hwa Lin; Koichi Kawakami; Tzu-Hsuan Yang; Jen-Leih Wu. Zebrafish eggs used as bioreactors for the production of bioactive tilapia insulin-like growth factors. Transgenic Research 2010, 20, 73 -83.

AMA Style

Shao-Yang Hu, Chia-Hsuan Liao, Yi-Pei Lin, Yen-Hsing Li, Hong-Yi Gong, Gen-Hwa Lin, Koichi Kawakami, Tzu-Hsuan Yang, Jen-Leih Wu. Zebrafish eggs used as bioreactors for the production of bioactive tilapia insulin-like growth factors. Transgenic Research. 2010; 20 (1):73-83.

Chicago/Turabian Style

Shao-Yang Hu; Chia-Hsuan Liao; Yi-Pei Lin; Yen-Hsing Li; Hong-Yi Gong; Gen-Hwa Lin; Koichi Kawakami; Tzu-Hsuan Yang; Jen-Leih Wu. 2010. "Zebrafish eggs used as bioreactors for the production of bioactive tilapia insulin-like growth factors." Transgenic Research 20, no. 1: 73-83.

Journal article
Published: 27 November 2009 in Marine Biotechnology
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Genetically modified (GM) fish with desirable features such as rapid growth, disease resistance, and cold tolerance, among other traits, have been established in aquaculture. However, commercially available GM fish are restricted because of global concerns over the incomplete assessments of food safety and ecological impact. The ecological impact concerns include gene flow and escape of the GM fish, which may cause extinction of wild natural fish stocks. Infertility control is a core technology for overcoming this obstacle. Although polyploidy technology, GnRH-specific antisense RNA, and RNAi against GnRH gene expression have been used to cause infertility in fish, these approaches are not 100% reliable and are not heritable. In the present study, zebrafish was used as a model to establish an inducible platform of infertility control in GM fish. Nitroreductase, which converts metronidazole substrate into cytotoxin, was fused with EGFP and expressed specifically by oocytes in the Tg(ZP:NTR-EGFP) by a zona pellucida promoter. Through consecutive immersion of metronidazole from 28 to 42 days posthatching, oocyte-specific EGFP expression was eliminated, and atrophy of the gonads was detected by anatomical analysis. These findings reveal that oocyte-specific nitroreductase-mediated catalysis of metronidazole blocks oogenesis and leads to an undeveloped oocyte. Furthermore, oocyte cell death via apoptosis was detected by a TUNEL assay. We found that the gonadal dysgenesis induced by metronidazole resulted in activation of the ovarian killer gene bok, which is a proapoptotic gene member of the Bcl-2 family and led to infertility. These results show that oocyte-specific nitroreductase-mediated catalysis of metronidazole can cause reliable infertility in zebrafish and could potentially be used as a model for other aquaculture fish species.

ACS Style

Shao-Yang Hu; Pei-Yu Lin; Chia-Hsuan Liao; Hong-Yi Gong; Gen-Hwa Lin; Koichi Kawakami; Jen-Leih Wu. Nitroreductase-mediated Gonadal Dysgenesis for Infertility Control of Genetically Modified Zebrafish. Marine Biotechnology 2009, 12, 569 -578.

AMA Style

Shao-Yang Hu, Pei-Yu Lin, Chia-Hsuan Liao, Hong-Yi Gong, Gen-Hwa Lin, Koichi Kawakami, Jen-Leih Wu. Nitroreductase-mediated Gonadal Dysgenesis for Infertility Control of Genetically Modified Zebrafish. Marine Biotechnology. 2009; 12 (5):569-578.

Chicago/Turabian Style

Shao-Yang Hu; Pei-Yu Lin; Chia-Hsuan Liao; Hong-Yi Gong; Gen-Hwa Lin; Koichi Kawakami; Jen-Leih Wu. 2009. "Nitroreductase-mediated Gonadal Dysgenesis for Infertility Control of Genetically Modified Zebrafish." Marine Biotechnology 12, no. 5: 569-578.

Conference abstract
Published: 31 August 2009 in Mechanisms of Development
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Shao-Yang Hu; Hong-Yi Gong; Jim-Ray Chen; Gen-Hwa Lin; Jen-Leih Wu. 01-P024 MicroRNA-206 regulates sonic hedgehog to control myogenesis. Mechanisms of Development 2009, 126, S58 -58.

AMA Style

Shao-Yang Hu, Hong-Yi Gong, Jim-Ray Chen, Gen-Hwa Lin, Jen-Leih Wu. 01-P024 MicroRNA-206 regulates sonic hedgehog to control myogenesis. Mechanisms of Development. 2009; 126 ():S58-58.

Chicago/Turabian Style

Shao-Yang Hu; Hong-Yi Gong; Jim-Ray Chen; Gen-Hwa Lin; Jen-Leih Wu. 2009. "01-P024 MicroRNA-206 regulates sonic hedgehog to control myogenesis." Mechanisms of Development 126, no. : S58-58.

Journal article
Published: 31 December 2008 in Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology
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Gonadotropin-releasing hormone (GnRH) is a neuropeptide that plays a major role in releasing pituitary gonadotropin and controlling vertebrate reproduction. In this study, three GnRH cDNAs, GnRH-I (sbGnRH; 348 bp), GnRH-II (cGnRH-II; 557 bp), and GnRH-III (sGnRH; 483 bp), were cloned from the brain of the silver sea bream (Sparus sarba). In order to understand how the expression of the GnRH isoforms was regulated in the brain, the promoter of each gene was cloned and analyzed. We found regulatory motifs in the promoters that were conserved in the GnRH promoters of tilapia and zebrafish, suggesting that these motifs play a critical role in GnRH regulation. We performed functional analyses and examined tissue-specific expression for each GnRH promoter using EGFP reporter fusions in zebrafish. The GnRH-I promoter was active in the forebrain area, including the olfactory bulb-terminal nerve area and peripheral preoptic areas; the GnRH-II promoter was active in the midbrain; and the GnRH-III promoter was active in the olfactory bulb. These results show that the GnRH promoters of the silver sea bream GnRH genes exhibit tissue-specific activity.

ACS Style

Shao-Yang Hu; Mark Hung-Chih Chen; Yi-Chia Lin; Gen-Hwa Lin; Hong-Yi Gong; Tzu-Hsuan Yang; Jen-Leih Wu. Cloning and functional analysis of the proximal promoter region of the three GnRH genes from the silver sea bream (Sparus sarba). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2008, 151, 373 -380.

AMA Style

Shao-Yang Hu, Mark Hung-Chih Chen, Yi-Chia Lin, Gen-Hwa Lin, Hong-Yi Gong, Tzu-Hsuan Yang, Jen-Leih Wu. Cloning and functional analysis of the proximal promoter region of the three GnRH genes from the silver sea bream (Sparus sarba). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology. 2008; 151 (4):373-380.

Chicago/Turabian Style

Shao-Yang Hu; Mark Hung-Chih Chen; Yi-Chia Lin; Gen-Hwa Lin; Hong-Yi Gong; Tzu-Hsuan Yang; Jen-Leih Wu. 2008. "Cloning and functional analysis of the proximal promoter region of the three GnRH genes from the silver sea bream (Sparus sarba)." Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 151, no. 4: 373-380.