Physiological, biochemical and HSP70 and HSP90 gene expression profiles of tropical abalone Haliotis squamata in response to Vibrio alginolyticus infection

Vibrio spp. have been known responsible for fish diseases in marine and brackish‐water systems in the tropics regions. Heat shock proteins are a highly conserved protein group that is known for its rapid response to environmental stresses, including infection. This study aimed to investigate physiological and biochemical responses of tropical abalone Haliotis squamata to Vibrio alginolyticus infection. Abalones were infected with V. alginolyticus by intramuscular injection at a dose of 105,106,107 cfu/abalone. The expression of HSP70 and HSP90 genes, the activity of superoxide dismutase, phenol oxidase and catalase enzymes, histology, falling and mortality were observed at 12, 24, 48, 72, and 96 hours post‐infection (hpi). The different expression of HSPs was found in this study. While the expression of HSP70 was downregulated after infection, the expression of HSP90 was upregulated at 12 hpi and followed by downregulated after 24 hpi for 106 cfu infection, but expressed at a normal level for 105 infection treatment. The expression ofsuperoxide dismutase and catalase increased within 12 hpi, and the expression of phenol oxidase increased after 24 hpi. V. alginolyticus is virulent with LD50 of less than 105 cfu on H. squamata with an average weight of 5.13 g, and caused enlargement of hemolymph sinus and development intraepithelial and intramuscular abscesses

COMPARISON OF THREE DIFFERENT TECHNIQUES OF GENE TRANSFER IN HUMPBACK GROUPER ( ) CROMILEPTES ALTIVELIS

Humpback grouper is one of the most cultured fishes in Asia, including Indonesia. Themain problemfaced by humpback culture is its slow growth rate.One of themethods that willbe more effective and efficient to solve the problem is using transgenic technique. This studywas conducted to determine the effectiveness of transfection,microinjection and electroporationtechniques on gene transfer in humpback grouper. transfection was performed byincubating sperm to the foreign DNA (pktBP-ktGH gene construct)-transfectant complexsolution, while was by injecting those complex solution into testis of mature males.Microinjection was conducted in 2-4 cell stage embryos using 25 μg/ml of foreign DNAsolution, and duration of injection was 1, 2 and 3 seconds. Electroporation by 50 V, 30 ms ofpulse length, 5 of pulse number and 0.1 of pulse interval was performed to sperm using threeDNA concentration of 5, 10 and 20 μg/ml. The incorporation of foreign DNA in sperm andembryos were analyzed using PCR method. Based on PCR analysis, an optimum DNAconcentration for electroporation was 10 μg/ml. Limited number of embryos could bemicroinjected during 20-30 min to reach 2-4 cell stage. Microinjection for 1 second showedhigher survival rate of embryos, although none or very low number of larvae was hatched.Transfast was an effective DNA delivery reagent for humpback grouper sperm. Foreign DNAcould be detected in sperm from two out of ten transfected fish at least 36 hours posttransfection (hpt). By transfection, foreign DNA was detected in sperm at 48 hpt 25 Cincubation temperature. Our study revealed that transfection, microinjection as well aselectroporation could be used as transgenesis methods in humpback grouper. By means ofsimplicity and efficacy, however, electroporationwas an appropriate gene transfermethod.oIn vitroin vivoin vivoin vitr

Comparison of Three Different Techniques of Gene Transfer in Humpback Grouper ( ) Cromileptes Altivelis

Humpback grouper is one of the most cultured fishes in Asia, including Indonesia. Themain problemfaced by humpback culture is its slow growth rate.One of themethods that willbe more effective and efficient to solve the problem is using transgenic technique. This studywas conducted to determine the effectiveness of transfection,microinjection and electroporationtechniques on gene transfer in humpback grouper. transfection was performed byincubating sperm to the foreign DNA (pktBP-ktGH gene construct)-transfectant complexsolution, while was by injecting those complex solution into testis of mature males.Microinjection was conducted in 2-4 cell stage embryos using 25 μg/ml of foreign DNAsolution, and duration of injection was 1, 2 and 3 seconds. Electroporation by 50 V, 30 ms ofpulse length, 5 of pulse number and 0.1 of pulse interval was performed to sperm using threeDNA concentration of 5, 10 and 20 μg/ml. The incorporation of foreign DNA in sperm andembryos were analyzed using PCR method. Based on PCR analysis, an optimum DNAconcentration for electroporation was 10 μg/ml. Limited number of embryos could bemicroinjected during 20-30 min to reach 2-4 cell stage. Microinjection for 1 second showedhigher survival rate of embryos, although none or very low number of larvae was hatched.Transfast was an effective DNA delivery reagent for humpback grouper sperm. Foreign DNAcould be detected in sperm from two out of ten transfected fish at least 36 hours posttransfection (hpt). By transfection, foreign DNA was detected in sperm at 48 hpt 25 Cincubation temperature. Our study revealed that transfection, microinjection as well aselectroporation could be used as transgenesis methods in humpback grouper. By means ofsimplicity and efficacy, however, electroporationwas an appropriate gene transfermethod.oIn vitroin vivoin vivoin vitr

Survailen Aktif Infectious Myonecrosis Virus (Imnv) pada Udang Vannamei (Litopenaeus vannamei) yang Dibudidayakan di Jawa Timur Dan Bali

An active surveillance of infectious myonecrosis virus (IMNV) in pond-cultured white shrimp (Litopenaeus vannamei) was conducted to determine the status of this disease. Samples of white shrimp (L. vannamei) were taken from ponds in East Java and Bali using proportional sampling. The sample was subjected to IMNV detection with Polymerase Chain Reaction (PCR) method and confirmed with histopathology. Results of this study showed that IMNV was found in shrimp samples from Kapongan, Situbondo district with the prevalence of 11,1%. Meanwhile, samples from other districts in East Java and Bali had no IMNV disease during this study. There is no treatment for viral disease in shrimp culture. Therefore, control of movement of live shrimp and application of biosecurity-area based should be more implemented among the shrimp culture area