Trad. Med. J., September-December 2019 Vol. 24(3), p 184-188 ISSN-p : 1410-5918 ISSN-e : 2406-9086 184 DOI: 10.22146/mot.45315 | Traditional Medicine Journal, 24(3), 2019 Submitted : 23-04-2019 Revised : 28-09-2019 Accepted : 20-11-2019 Antibacterial Activity of Cuttlefish Sepia sp. (Cephalopoda,) Ink Extract Against Aeromonas hydrophila R Adharyan Islamy * Departement of Aquaculture, Faculty of Fisheries and Marine Science, Brawijaya University Malang ABSTRACT Cephalopods ink has shown potential antiretroviral activity. The ink extracts of cuttlefish showed an antibacterial effect. This study aims to investigate the antibacterial activity of the methanolic extract of the ink of cuttlefish Sepia sp. against Aeromonas hydrophila. A. hydrophila are opportunists and associated with aquatic fish and shrimp disease. The shade-dried ink sample from approximately 30g ink sacs obtained from 15 animals was immersed separately in methanol (1:3 w/v) solvents for overnight. The experiment in this study used the dried extract of cuttlefish ink. The isolate of A. hydrophila was originated from Jepara Brackishwater Aquaculture Center. The average yield percentage of cuttlefish ink extract obtained was 4.86%. The results of the MIC test in Table V show that the highest average absorbance value obtained was at a concentration of 50 ppm, which was equal to 1.716 nm, and the lowest absorbance was obtained at a treatment dose of 300 ppm at 0.841 nm, while the Mc Farland tube was 0.933 nm. The results of antibacterial test on Table II showed antibacterial activity of cuttlefish ink extract at negative concentration control showed diameter zone of 5 ± 1.2 mm, at positive control showed diameter zone of 31 ± 1.2 mm, at 250 ppm result 19 ± 0.9 mm, at 300 ppm result 22 ± 1.4 mm, at 350 ppm result 31 ± 1.2 mm. Keywords: A. hydrophila; Antibacterial; Cuttlefish Ink; Extract; Sepia sp. INTRODUCTION A. hydrophila is a gram-negative opportunist bacterium associated with aquatic animal disease (Barker, 2001). A. hydrophila causes mass mortalities in several species, including Carps, Snakeheads, Gouramies and Catfishes and is considered as an etiological agent of several diseases such as emaciation, hemorrhagic septicemia, asymptomatic septicemia, ulcerative infection, tail rot and fin rot (Rahman et al., 2001). It also causes Motile Aeromonas Septicaemia (MAS) and Epizootic Ulcerative Syndrome (EUS) as a primary pathogen (Roberts et al., 1993). The use of antimicrobials that are not as recommended for disease control in human beings and animals has increased the natural emergence of bacterial resistance (Allsop, 1998). The emergence of multiple resistance has dramatically decreased the effectiveness of the antibiotics, mostly. Therefore, searching for novel antibacterial compounds with therapeutic potential for which the pathogens may not have resistance is necessary (Patil et al., 2001). The Cephalopods classes come under the phylum Mollusca, which involves squid, cuttlefish, octopuses, and nautilus (Voss, 1973). The cephalopods live in all marine habitats and are famous for their defenses, from their fast getting escape movements to changes in coloration that *Corresponding author: R Adharyan Islamy Email: r.adhariyan@gmail.com can be disruptive, cryptic, or startling, to arm autotomy, to toxin venom and inking (Hanlon and Messenger, 1996; Normal, 2000). Cephalopod ink has shown potential antiretroviral activity (Rajaganapathi et al., 2000). Squid and cuttlefish release ink from their ink sac to escape from their predators (Ortonne et al., 1981). The ink extracts of cuttlefish showed antibacterial effect (Nithya et al., 2011; Vennila et al., 2011; Zainab and Abas, 2010; Annaian et al., 2008; Shanmugam et al., 2008). The aim of this study was to analyze the antibacterial activity of the cuttlefish ink Sepia sp. extract against A. hydrophila. METHODOLOGY Extracts preparation Wash the cuttlefish Sepia sp. using sterile water. Then removing the ink sacs by dissections. Squeeze out the cuttlefish sac gently until the ink release. The shade-dried ink sample from approximately 30g ink sacs obtained from 15 animals was immersed separately in methanol (1:3 w/v) solvents for overnight. The extracts then filtered through the Whatman No.1 filter paper. The filtrate was poured in previously weighed Petri dishes, evaporated to dryness, and the dried extract was used for the experiments. The yield of Cuttlefish Ink Extract Sepia sp. The yield analysis of ink extract based on a comparison of bag weight (containing ink) samples to the weight of the extract of the ink container.