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Characteristics of Salted Catfish (Pangasius hypophthalmus) with Salt Concentration Variations and Time of Fermentation INFLUENCE OF ISOTROPIC GRAVITY CULTURE Merkuria Karyantina 1, Yustina Wuri Wulandari 1, and Fitri Kharismawati 1 ON CYTOSKELETON STRUCTURE AND FORMATION OF FOCAL ADHESIONS IN HUMAN 1Faculty of Technology and Food Industry, Slamet Riyadi University, Surakarta MESENCHYMAL STEM CELLS E-mail: [email protected] C. Koaykul1, M. Kim1, Y. Kawahara2, L. Yuge2, 3, M. Kino-oka1 Fish protein has a privilege that besides more digestible and also contains amino 1Department of Biotechnology, Osaka University, Osaka, Japan acids which had a pattern similar to the pattern of amino acids in the human body. 2Space Bio-Laboratories Co., Ltd., Hiroshima, Japan One of the many types of fish consumed by people is catfish (Pangasius 3Division of Bio-Environmental Adaptation Sciences, Hiroshima University, Hiroshima, Japan hypophthalmus). Catfish is easily damaged so must be preserved using NaCl. The process of decay in fish can be caused mainly by the activity of enzymes found in A three-dimensional clinostat is a device for generating isotropic gravity culture with an the body of the fish itself, the activity of microorganisms, or the oxidation process average of 10−3 G over time. This present study aimed to understand effects of disperse in the fat body by oxygen from the air, in addition the fungus also causes damage gravity vector on actin cytoskeleton structure and formation of focal adhesions in to the fish. The study evaluated the effect of salt concentration and time of human mesenchymal stem cells (hMSCs). By comparing the cultures between fermentation on the characteristics of salted fish. The research method with a unidirectional gravity condition and isotropic gravity condition, we found that hMSCs completely randomized design with 2 factors, the concentration of salt (20, 30, and in unidirectional gravity condition showed actin stress fibers appeared with longitudinal 40%) and time of fermentation (24, 36 and 48 h). Analysis is conducted on direction along the cells, which located both of apical and basal sides of the nucleus. On moisture, ash, protein content, total plate count, total mold and sensory evaluation the other hand, hMSCs in isotropic gravity condition presented broader lamellipodia test. Research showed that the best treatment in this study was in treatment salt and the stress fibers located mainly at basal side but not apical side of the nucleus. content of 30% and fermentation time 24 h. Characteristics of these treatments are Furthermore, paxillin, focal adhesion protein, was detected in the peripheral and central 14.16% moisture content, ash content of 2.87%, 5.73% protein content and regions of the cells in both culture conditions, especially at the ends of the stress fibers organoleptic test results saltiness (3.00/saltiness), crispness (1.60), color (2.50 with intensive staining in the longitudinal direction in a scattered manner. Interestingly, brown) and the overall favorite (2.90). Total plate count 1.1 x 105 CFU/g and total phosphorylated-paxillin spots were discovered in the periphery of the cells cultured in mold 7.2 x 102 CFU/g. isotropic gravity condition. However, phosphorylated-paxillin was not observed in Keywords: salted; catfish; fermentation; microorganism hMSCs cultured in unidirectional gravity condition. In conclusion, this study revealed that isotropic gravity culture effects on actin cytoskeleton structure and turnover of focal adhesions in hMSCs relating to migration activity and mechanotransduction in the cells. INTRODUCTION _________________________________ Fish has been commonly used as a source of animal protein. Fish protein has a Abstract of ISCT 2018 Montreal Annual Meeting in Canada organized by International Soc. for privilege that is easy to digest and contains amino acids with a pattern similar to the Cell & Gene Therapy, Poster No.113. pattern of amino acids in the human body. Advantages of utilizing fish meat as a protein Chaiyong Koaykul: Participant of the 3rd UO, 2013-2014. source is high protein (20%), containing slightly woven binder so easily digested, although has high of fatty acid (0.1 - 2.2%) but 25% of this amount is an acid- unsaturated fatty acids that humans need, cholesterol levels are very low, contains a number of minerals (K, Cl, P, S, Mg, Ca, Fe, Zn, F, Ar, Cu and Y), vitamins A and D in an amount sufficient for the human body. The fish meat can be accepted by all levels of society. Weakness of a fish's body has a high water content (80%) and pH close to neutral so it is a medium for growing mold, spoilage bacteria and other microorganisms. Meat fish is susceptible to autolysis so that the meat becomes soft. The means often used to preserve fish are temperature modification (canning, refrigeration and freezing), chemical means (use of salt and vinegar) and drying (natural Received August 30, 2018 / Accepted October 22, 2018. Merkuria Karyantina: Participant of the 3rd UO, 2013-2014. 382 383 and artificial). Salting techniques there are two types, namely dry and wet salting (Buckle, 1987). One of the many fishes consumed by people is catfish (Pangasius hypophthalmus). Catfish is quite easy to find on the market, so it can become an alternative source of animal protein. As with other fish products, catfish is easily damaged so should need the effort to preserve. In fermentation need salt, which plays a role in the decomposition of the compound is the enzyme of the fish, especially from the contents of the stomach and microorganisms from fish. Bacteria that develop during fermentation with salt are mainly from the type of Micrococcus, Bacillus, and Sarcina which have halotoleran nature. High salt levels in the product, capable of inhibiting Staphylococcus aureus. Salt causes the withdrawal of water from the food (fish) so Aw fish will be declined and the growth of some microorganisms will be inhibited (Rahayu, et al. 1992). Combination of pH and salt in many fermentation products tends to inhibit the growth of microorganisms. During fermentation, pH is reduced below 5 and safe processing depends on proper salt content, temperature control, and pH (Fernandes, 2009). This study will assess the effect of salt concentration and fermentation time on the quality of salted fish (jambal) catfish, to avoid microorganism and mold growth. The concentration of salt used is 20, 30, and 40%. Factors used fermentation time is 24, 36 and 48 h. Salted fish products will be analyzed levels of protein, moisture content, ash content and microbiology analysis (Total plate count and total mold). METHODS The design used was completely randomized design (CRD) with 2 factors. Factor 1: the concentration of salt (G1: 20%, G2: 30%, G3: 40%). Factor 2: time of fermentation (F1: 24 h, F2: 36 h, F3: 48 h). The data obtained by analysis of variance (ANOVA) with a significant level of 5%. Then, the calculation result is no real difference followed by Tukey's test. Materials research is catfish, salt crystals, chemicals for analysis is distilled water and chemicals for ash, water content (AOAC, 1992) and protein analysis (Mikro Kejhldal method), microbiology analysis (Total Plate Count using Plate Count Agar + 5% NaCl and Total mold using Potatoes Dextrose Agar + 5% NaCl) and sensory evaluation (with hedonic test). Preparation of salted fish: fish cleaned and cut of visceral fish, cut with a thickness of 3cm and added crystal salt. Then fermented for 24, 36, 48 h and washed with fresh water. Fish dried for 3-4 d, with the cabinet dryer. For sensory evaluation, fermented fish fried with hot oil for 3 min. RESULTS AND DISCUSION a. Chemical Analysis The water content of salted fish catfish showed a tendency to increase along with the high salt concentration and fermentation time. According (Fernandes, 2009) that the salting process will stop after a proper balance between the solution in the fish meat with a salt solution on the outside during certain salting. Rochima (2005) stated that during the process of salting will happen salt penetration into the body of the fish 384 385 and artificial). Salting techniques there are two types, namely dry and wet salting depends on the purity of the salt used. Some important factors that influence the (Buckle, 1987). effectiveness of salting are salt concentration, temperature salting, the thickness of the One of the many fishes consumed by people is catfish (Pangasius fish meat and the level of freshness of the fish. The length of time determined by the hypophthalmus). Catfish is quite easy to find on the market, so it can become an speed of salting salt dissolves to form 'brine', the speed of the salt into the flesh of fish alternative source of animal protein. As with other fish products, catfish is easily and attract water, the amount of salt or brine density, temperature salting, size of the fish. damaged so should need the effort to preserve. In fermentation need salt, which plays a role in the decomposition of the Table 1. Chemical analysis compound is the enzyme of the fish, especially from the contents of the stomach and Salt concentration Time of fermentation Water Ash (%) Protein (%) microorganisms from fish. Bacteria that develop during fermentation with salt are (%) (h) content (%) 20 24 7.35 3.14 5.89 mainly from the type of Micrococcus, Bacillus, and Sarcina which have halotoleran nature. High salt levels in the product, capable of inhibiting Staphylococcus aureus. Salt 36 8.66 2.73 5.83 48 12.54 2.36 5.51 causes the withdrawal of water from the food (fish) so Aw fish will be declined and the 30 24 14.16 2.87 5.73 growth of some microorganisms will be inhibited (Rahayu, et al. 1992). 36 10.50 2.82 5.34 Combination of pH and salt in many fermentation products tends to inhibit the 48 15.63 2.20 5.41 growth of microorganisms.
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