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All Journal Jurnal Akuakultur Indonesia Indonesian Aquaculture Journal
Mira Mawardi
Balai Besar Pengembangan Budidaya Air Tawar, Sukabumi. Jl. Selabintana No. 37 Sukabumi 43114
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

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Lactococcus garvieae: CHARACTERIZATION AND ABILITY TO INHIBIT THE GROWTH OF AQUACULTURE PATHOGENIC BACTERIA Mawardi, Mira; Indrawati, Agustin; Lusiastuti, Angela Mariana; Wibawan, I Wayan Teguh
Indonesian Aquaculture Journal Vol 19, No 1 (2024): (June, 2024)
Publisher : Agency for Marine and Fisheries Extension and Human Resources

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/iaj.19.1.2024.87-98

Abstract

Lactococcus garvieae is a gram-positive ovoid cocci bacterium formerly classified as a member of the Lactococcus genus. This study aims to isolate L. garvieae from catfish rearing pond and characterize it as a potential probiotic candidate. L. garvieae was identified and characterized through phenotypic and genotypic observation, genomic % G~C content analysis, cell surface hydrophobicity assays, acidification test, in vitro antagonism, and a profile of antimicrobial activities. The MT597595.1 accession number corresponds to L. garvieae, as determined by a molecular identification test. Biochemical characterization was performed using API 50 CH kit. The genomics %G~C content of L. garvieae was 51.8. Findings from acidification ability tests, in vitro antagonism tests, and the ability of bacteria to grow in broth medium at pH 4 reveal that L. garvieae can inhibit the growth of Aeromonas hydrophila, Streptococcus agalactiae, Streptococcus iniae, and Edwardsiella ictaluri. However, it does not suppress the growth of L. garvieae  Edwardsiella tarda. Remarkably, L. garvieae has the ability to reduce the pH of neutral broth medium turning it acidic. Furthermore, L. garvieae’s hydrophobic cell surface exhibited an adhesive, hydrophobic, and protein surface cell content with a compact growth pattern consistent with postive SAT and MATH assay. Antimicrobial activity tests, encompassing 11 antibiotics, disclosed resistance to  Nalidixic acid while displaying intermediate sensitivity to Streptomycin and Trimethoprim. In conclusion, L. garvieae demonstrates an inhibitory effect on the growth of pathogenic bacteria, underlining its potential as a probiotic candidate.
CHARACTERIZATION OF SPORE-FORMING BACTERIA ISOLATED FROM TILAPIA (OREOCHROMIS NILOTICUS) AND THEIR POTENTIAL FOR A PROBIOTIC CANDIDATE Mawardi, Mira; Indrawati, Agustin; Lusiastuti, Angela Mariana; Wibawan, I Wayan Teguh
Indonesian Aquaculture Journal Vol 18, No 2 (2023): (December, 2023)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/iaj.18.2.2023.105-114

Abstract

Gram-positive spore bacteria are widely used as probiotics in general sectors. However, there are still limited bacterial isolates as probiotic candidates available from indigenous isolates, especially in aquaculture. This study aimed to obtain potential spore-forming isolates as probiotic candidate for tilapia. Tilapia fish samples were collected from Sukabumi, Ciamis, Serang, and Papua. Bacterial isolates were isolated from the digestive tract of tilapia. Bacteria were identified based on their morphological, molecular characteristics, complete genome composition, and cell surface identification based on hydrophobic properties. In this study, six bacteria were isolated and identified by molecular characteristics using 16S rRNA sequences. Based on the phylogenetic analysis, the 9 PP isolate was Priestia megaterium basonym: Bacillus megaterium, CMS 16N isolate was Brevibacillus halotolerans, PPN 10 isolate was Bacillus sp., 3.1 SKBM isolate was Bacillus mycoides, CMS 22 N and SRG32 isolate were Bacillus subtilis. Six bacteria had different phenotypicals, ATGC sequence compositions, and a higher proportion of total G~C sequence composition above 50%. The coherent cell surface hydrophobicity test was positive on the SAT, SA, AA, and compact growth patterns in soft-agar media for 9 PP, CMS 22 N, and SRG32 isolates. From our study, the indigenous spore-forming bacteria isolated from tilapia stomachs are enzymatic bacteria, which have a strong attachment to host tissue and high potential as a probiotic candidate for fish. Various hydrophobicity test results from each isolate indicate that the protein composition in the cell surface is different.
Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion Nuswantoro, Soko; Alimuddin, .; Yuhana, Munti; Santika, Ayi; Nuryati, Sri; Zainun, Zakki; Mawardi, Mira
Jurnal Akuakultur Indonesia Vol. 11 No. 1 (2012): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (991.677 KB) | DOI: 10.19027/jai.11.76-85

Abstract

Koi herpesvirus (KHV) is a herpesvirus that particularly infects and causes mass mortality to koi and common carp. Therefore, the protection of common carp from KHV infection is urgently needed. In this study, we developed an application of DNA vaccine encoding KHV glycoprotein-25 by immersion method to increase survival of common carp against KHV infection. A total of 400 common carp juveniles at 30-day-old were immersed in 1-L water containing 1.3×108CFU/mL of the killed Escherichia coli cells carrying DNA vaccine. Three frequencies and three duration of fish immersion were tested, namely: 1×30 minutes, 1×60 minutes, 1× 90 minutes, 2×90 minutes and 3×90 minutes by interval of 24 hours. Reversetranscription polymerase chain reaction analysis showed that DNA vaccine was successfully expressed in the vaccinated fish. Fish at twenty eight days post vaccination were challenged by injecting 10-4 mL of KHV per fish. The result showed that vaccination by 1×30 minutes immersion allowed 61% of fish survived, and this was significantly higher (p<0.05) compared to control (without vaccination), but it was similar among vaccination treatments (p>0.05). The relative percent survival of vaccinated fish were also similar among treatments (p>0.05). DNA vaccination has increased fish survival about two fold higher compared to unvaccinated fish control (26.67%). Thus, DNA vaccination was effectively delivered by immersion for 1×30 minutes, and this technique can be useful to level up the resistance of common carp juveniles against KHV infection. Keywords: DNA vaccine, KHV, glycoprotein, immersion, common carp
Pathogenicity test bacteria from Oreochromis niloticus and Clarias gariepinus aquaculture ponds Hardi, Esti Handayani; Halim, Aldi M.; Nugroho, Rudy Agung; Mawardi, Mira; Isnansetyo, Alim; Mariana Lusiastuti, Angela; Rahayu, Widianingsih; Niklani, Andi; Saptiani, Gina
Jurnal Akuakultur Indonesia Vol. 22 No. 1 (2023): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19027/jai.22.1.97-105

Abstract

Present research aimed to examine the pathogenicity opportunities of some bacteria which were isolated from freshwater aquaculture ponds to some fish Oreochromis niloticus, Clarias gariepinus, and Pangasius pangasius. The bacteria were isolated from the water and feces of O. niloticus and C. gariepinus aquaculture in Samarinda, which were identified as Escherichia coli, Enterobacter cloacae, and Enterobacter amnigenus. An 0.1 mL-1 of each bacteria was intramuscular injection and evaluated the clinical signs, eksternal pathology, and fish mortality. The freshwater fish using in this research were Oreochromis niloticus, Clarias gariepinus, and Pangasius pangasius with size 10-15 g from Loa Kulu, Kutai Kartanegara Regency, East Kalimantan, Indonesia. The result showed that each bacteria caused mortality in fish: E. coli bacteria caused fish mortality 23.33-66.67%; E. cloacae 10.00 to 90.00%; and E. amnigenus by 3.33-56.67%. The average death time of E. coli and E. amnigenus in all three fish were under 48 hours, while E. cloacae caused mortality less than 48 hours in C. gariepinus and P. pangasius. In conclusion, E. coli, E. cloacae, and E. amnigenus are putative pathogenic bacterium in O. niloticus, C. gariepinus, and P. pangasius.
Co-Authors Agustin Indrawati ALIM ISNANSETYO Alimuddin ANGELA MARIANA LUSIASTUTI Angela Mariana Lusiastuti Ayi Santika Esti Handayani Hardi Gina Saptiani Halim, Aldi M. I wayan Teguh Wibawan MUNTI YUHANA Niklani, Andi Rahayu, Widianingsih Rudy Agung Nugroho Soko Nuswantoro Sri Nuryati Zakki Zainun