Molecular identification of bacteria isolated from culture medium of rotifer fed on fishery waste diet
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Abstract. Wullur S, Napitupulu H, Wantania LL, Ginting EL, Warouw V, Tallei TE, Rumengan IFM. 2020. Molecular identification of bacteria isolated from culture medium of rotifer fed on fishery waste diet. Biodiversitas 21: 2735-2740. The aim of this study was 16S-rRNA sequences based molecular identification of bacteria isolated from culture medium of rotifer fed with fishery waste diet (FWD). We cultured rotifer Brachionus rotundiformis in sterilized seawater (salinity 25 ppt) using FWD, following the procedure in Patent No. P00201609066. Bacteria from the culture were collected, homogenized, diluted 10 to 1000 fold, spread on agar plates and incubated at 370C for 24 to 48 hours. Representative colonies of the bacteria according to their morphologies were isolated for further characterization. Genomic DNA of the isolates were extracted, and the 16S rRNA gene of the isolates were amplified. Polymerase Chain Reaction (PCR) product of each isolate was sequenced and queried against the NCBI GenBank database. Six different isolates based on size, color, elevation, margin, and colony were observed during 24-48 hours incubation at 370C. The 16S rRNA genes of the six isolates were successfully amplified and produced DNA band at 1300-1500 bp, with quality value equal to or greater than 20 (QV20+) of each entire sequence around 941-1253 bases. Basic Local Alignment Search Tool (BLAST) queries in the NCBI GenBank and EzBioCloud database using the 16S-rRNA gene sequences showed that the six isolates belong to four different genera, i.e: Bacillus, Staphylococcus, Vibrio, and Alteromonas.
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References
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Gallardo, W. G., Hagiwara, A., Tomita, Y., Soyano, K., and Snell, T. W. 1997. Effect of some vertebrate and invertebrate hormones on the population growth, mictic female production, and body size of the marine rotifer Brachionus plicatilis Müller. Hydrobiologia, 358(1-3), 113-120.
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Gomez, B., Thompson, F. L., Thompson, C. C., and Swings, J. 2003. Vibrio rotiferianus sp. nov., isolated from cultures of the rotifer Brachionus plicatilis. International Journal of Systematic and Evolutionary Microbiology, 53(1), 239-243.
Haché, R., and Plante, S. 2011. The relationship between enrichment, fatty acid profiles and bacterial load in cultured rotifers (Brachionus plicatilis L-strain) and Artemia (Artemia salina strain Franciscana). Aquaculture, 311(1-4), 201-208.
Hagiwara, A., Kim, H. J., and Marcial, H. 2017. Mass culture and preservation of Brachionus plicatilis sp. complex. In: Hagiwara A, Yoshonaga T. (eds) Rotifers aquaculture, ecology, gerontology, and ecotoxicology. Springer, Singapore.
Hagiwara, A., Kotani, T., Snell, T. W., Assava-Aree, M., and Hirayama, K. 1995. Morphology, reproduction, genetics, and mating behavior of small, tropical marine Brachionus strains (Rotifera). Journal of experimental marine biology and ecology, 194(1), 25-37.
Hagiwara, A., Snell, T. W., Lubzens, E., and Tamaru, C. S. 1997. Live food in aquaculture. Developments in Hydrobiology, 124, 328.
Hagiwara, A., Gallardo, W. G., Assavaaree, M., Kotani, T., and De Araujo, A. B. 2001. Live food production in Japan: recent progress and future aspects. Aquaculture, 200(1-2), 111-127.
Hagiwara, A., Suga, K., Akazawa, A., Kotani, T., and Sakakura, Y. 2007. Development of rotifer strains with useful traits for rearing fish larvae. Aquaculture, 268(1-4), 44-52.
Hagiwara, A., Wullur, S., Marcial, H. S., Hirai, N., and Sakakura, Y. 2014. Euryhaline rotifer Proales similis as initial live food for rearing fish with small mouth. Aquaculture, 432, 470-474.
Hagiwara, A., Hamada, K., Hori, S., and Hirayama, K. 1994. Increased sexual reproduction in Brachionus plicatilis (Rotifera) with the addition of bacteria and rotifer extracts. Journal of Experimental Marine Biology and Ecology, 181(1), 1-8.
Hagiwara, A., Balompapueng, M. D., Munuswamy, N., and Hirayama, K. 1997. Mass production and preservation of the resting eggs of the euryhaline rotifer Brachionus plicatilis and B. rotundiformis. Aquaculture, 155(1-4), 223-230.
Hansen, G. H., Strøm, E., & Olafsen, J. A. 1992. Effect of different holding regimens on the intestinal microflora of herring (Clupea harengus) larvae. Appl. Environ. Microbiol., 58(2), 461-470.
Haryanti, H., Sugama, K., Tsumura, S., & Nishijima, R. 2017. Enhance production of black tiger shrimp Penaeus monodon postlarvae by probiotic bacterium Alteromonas sp. Indonesian Fisheries Research Journal, 7(1), 1-6.
He, R. P., Feng, J., Tian, J. L., Dong, S. L., and Wen, B. 2017. Effects of dietary supplementation of probiotics on the growth, activities of digestive and non-specific immune enzymes 8 in hybrid grouper (Epinephelus lanceolatus and Epinephelus fuscoguttatus). Aquac. Res., 48: 5782–5790.
Hirata, H., and Mori, Y., 1967. Culture of rotifer Brachionus plicatilis fed on baker’s yeast. Saibai Gyogyo 5,36–40 (in Japanese)
Hirata, H., 1980. Culture methods of the marine rotifer, Brachionus plicatilis. Min. Rev. Data File Fish. Res. Kagoshima Univ. 1, 27-46
Hirata, H., Murata, O., Yamada, S., Ishitani, H., & Wachi, M. 1998. Probiotic culture of the rotifer Brachionus plicatilis. In Rotifera VIII: A Comparative Approach (pp. 495-498). Springer, Dordrecht.
Hirayama, K. 1987. A consideration of why mass culture of the rotifer Brachionus plicatilis with baker’s yeast is unstable. In Rotifer Symposium IV (pp. 269-270). Springer, Dordrecht.
Hirayama, K., Maruyama, I., and Maeda, T. 1989. Nutritional effect of freshwater Chlorella on growth of the rotifer Brachionus plicatilis. In Rotifer Symposium V (pp. 39-42). Springer, Dordrecht.
Ito T, 1960. On the culture of mixohaline rotifer Brachionus plicatilis O. F. Müller in the sea water. Rep Fac Fish Prefectural Univ Mie 3:708–740. (in Japanese).
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