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Profile of Microflora “Jambal Roti” (Traditional Fermented Fish) with Pediococcus Sp (Pediococcus Acidilactici F11 and Pe

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Profile of Microflora “Jambal Roti” (Traditional Fermented Fish) with Pediococcus Sp (Pediococcus Acidilactici F11 and Pe Profile of Microflora “Jambal Roti” (Traditional Fermented Fish) with Pediococcus sp (Pediococcus acidilactici F11 and Pediococcus halophillus FNCC-0032) Aplication and 25% NaCl Merkuria Karyantina1,2,, Sri Anggrahini3, Tyas Utami34 and Endang S Rahayu34 1 Faculty of Technology and Food Industry, Slamet Riyadi University, Sumpah Pemuda Street No 18, Joglo, Kadipiro, Surakarta 2 Doctoral Program in Agricultural Technology, Gadjah Mada University, Flora Street No 1, Bulaksumur, Caturtunggal, Yogyakarta 3Faculty of Agricultural Technology, Gadjah Mada University, Flora Street No 1, Bulaksumur, Caturtunggal, Yogyakarta 4Center for Food and Nutrition Studies, Gadjah Mada University, Yogyakarta, Indonesia 2Corresponding author: [email protected] and [email protected] ABSTRACT “Jambal roti” is a fermented fish product from manyung fish, which is quite famous in Java. The term “jambal roti” refers to the salting and drying of fish. Manyung fishes (Arius thalassinus) are easily damaged so that they need to be preserved by salting. Traditional production uses 30% salt, so the product is too salty. Decreased use of salt, allows the development of pathogenic bacteria. This study examined the effect of NaCl concentration (25%) on microflora profile during making of “jambal roti”. The results showed that the total lactic acid bacteria in de Mann Rogosa and Sharpe medium had an increase (2 log cycles) in all aplication. Total bacteria in Plate Count Agar medium and Total enterobacteriaceae (in VRBA medium) tends to be stable. Total Salmonella- Shigella pathogen in SSA media tends decrease (3 log cycles). The data shows that Pediococcus sp is able to grow up to 25% salinity and suppressed the growth of Salmonella-Shigella. Salmonella-Shigella growth can be suppressed by Pediococcus sp when NaCl levels are lowered during manufacture. Key words : “jambal roti”, microflora profile, Pediococcus sp INTRODUCTION Fish has been commonly used as a source of animal protein. Fish protein has a privilege that is easily digested and contains amino acids with a pattern similar to the pattern of amino acids in the human body. Advantages of utilizing fish meat as a protein 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), _______________________________ Received 29 June 2020 / Accepted 30 September 2020 in ICBiotech. Merkuria Karyantina: Participant of the 3rd UO, 2013-2014. 474 475 vitamins A and D in an amount sufficient for the human body. The fish meat can be accepted by all levels of society (Buckle, 1987). Weakness of a fish's body has a high water content (80%) and pH close to neutral so it is a medium growing mold, spoilage bacteria and other microorganisms. Meat fish is susceptible to autolysis so that the meat becomes soft and become a medium for growth of microorganisms and mold. The means used to preserve fish are often used temperature modification (canning, refrigeration and freezing), chemical means (use of salt and vinegar) and drying (natural and artificial). For salting techniques there are two types, namely dry and wet salting (Buckle, 1987). Salt plays a role in the breakdown of compounds in fish, especially from viscera, in addition to the role of microorganisms from fish in the fermentation process. Bacteria that play a role in the fermentation process of fish include the types of Micrococcus, Bacillus, and Sarcina which are resistant to high salt conditions. The use of high salt in fermented fish products can suppress the growth of Staphylococcus aureus. Salt causes the withdrawal of water from the food (fish) so water activity (Aw) of 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) Pediococcus sp is a group of bacteriocin-producing homofermentative lactic acid bacteria (Pediocin) was isolated from fermented products. These bacteria have ability to preserve food because they can produce lactic acid which can reduce the pH media and produce bacteriocin (Pediocin), so that its presence suppresses the growth of other bacteria. P. acidilactici F-11 bacteria have been used in the production of salted fish (inasua) and are able to suppress the growth of coliform bacteria and increase the content of lactic acid bacteria (Nendisa and Rahayu 2001). The optimum temperature for Pediococcus growth is 25-30℃ and the optimum pH is ± 6. Species and strains of this genus differ in their tolerance or resistance to oxygen, pH, temperature, antibiotic resistance, and NaCl. “Jambal Roti ”is a well-known fermented fish product in Java, especially the north coast. Jambal Roti made in the traditional way is easily contaminated by pathogenic bacteria, physical and chemical contaminants. The salting process can suppress the growth of pathogenic bacteria, but still allows the growth of high salt resistant pathogenic bacteria. Traditional production uses 30% salt, so the product is too salty Meat and processed meat products are the preferred habitat for some Pediococcus strains, for example sausages and ham. When growing on meat, Pediococcus can produce diacetyl which acts as an antimicrobial, but can also eliminate the taste of food even in small amounts. The genus Pediococcus is widely involved in fermentation of plant parts, including P. acidilactici, P. dextrinicus, P. inopinatus, P. parvulus, and P. pentosaceus. Thongthai et al (1992) stated that many bacteria are found in fermented fish, including Bacillus, Micrococcus, Staphylococcus, Streptococcus, Pediococcus, and extremely halophilic red archaea, halophilic lactic acid bacteria, namely Tetragenococcus. 474 475 This study examined the effect of NaCl concentrations (25%) and Pediococcus sp (Pediococcus acidilactici F11 and Pediococcus halophillus FNCC-0032) on microflora profile in the making of “jambal fish” (Arius thalassinus). METHODS The experiment with aplication of Pediococcus sp (Pediococcus halophillus FNCC-0032 and Pediococcus acidilactici F-11) and 25 % NaCl. Pediococcus halophillus FNCC-0032 and P. acidilactici F-11 were purchased from FNCC (Food Nutrition and Culture Colection) Gadjah Mada University. Lactic acid bacteria was inoculated in de Mann Rogosa and Sharpe for 24 hour. Manyung fishes (Arius thalassinus) was purchased from PT Holy Mina Jaya Rembang, Central Java and stored in freezer box until used. Three experiment groups were set 600 gram of fish, and added salt was 25% during three days. Salt was purchased from local market. After salted, fish was washed with water and inoculated LAB culture (soaked) for 5 minute. P. halophillus FNCC-0032 and P. acidilactici F-11 were inoculated at 106CFU/ml for each the experiment. Then fishes were dried with sun drying method for 3 days. Every day, 10-15 gram each experiment was picked up for analysis. Lactic acid bacteria was analyzed with de Mann Rogosa and Sharpe (MRS) agar media. Total bacteria were analyzed with Plate Count Agar (PCA). Enterobacteriaceae was analyzed with Violet Red Bile Agar (VRBA) media and Salmonella-Shigella analysis with Salmonella-Shigella Agar (SSA) media. RESULT AND DISCUSION 1. Lactic acid bacteria profile on Jambal Roti Lactic acid bacteria are bacteria that play an important role in the fermentation process. The results of observations on the content of lactic acid bacteria during the fermentation process were shown in Figure 1. Figure 1. Growth of Lactic acid bacteria on MRS-agar (log cfu/gram) during fermentation of jambal roti 476 477 The result showed that total lactic acid bacteria was increase 2 log cycle/gram during fermentation of “jambal roti”. In the control treatment, lactic acid bacteria was increased, showing that ”jambal roti” fermentation has indegenous lactic acid bacteria that play a role during fermentation. The number of bacteria on MRS media has increased, because gram positive bacteria (lactic acid bacteria) that are resistant to high salt can survive. According to Rahayu (2003), lactic acid bacteria found in the fermented fish is Lactobacillus plantarum, L. curvatus, L. murinus and Streptococcus thermophilus L. vitulinus and P. damnosus play a role in fermentation of “jambal roti” from Pangandaran (Indriati, 2006). Pediococcus sp in this study plays a role in degradation of the components in fish into simpler components, for example the breakdown of proteins into amino acids. Pediococcus sp is able to survive in conditions of high salt content (25%) during the fermentation process. Figure 1 showed that at the end of the treatment, P. acidilactici was able to survive more than the control treatment and P. halophillus treatment. pH (Figure 2) during the fermentation process ranges from 4-6 where in the pH of the pathogenic bacteria was dies, but Pediococcus sp can still survive. Pediocin produced by Pediococcus sp during the fermentation process can suppress the growth of pathogenic bacteria such as Enterobacteriaceae and Salmonella-Shigella. 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). Figure 2. pH during fermentation of jambal roti 2. Total bacteria profile from “jambal roti” Total bacteria in “jambal roti” analyzed with Plate Count Agar (PCA) media. Figure 3 shows that the total number of bacteria decreased (1 log cycle) at the end of the treatment. The decrease can be caused by the number of pathogenic bacteria (Enterobacteriaceae and Salmonella-Shigella) which are suppressed by Pediocin form Pediococcus sp. 476 477 Figure 3. Growth of Total bacteria on Plate Count Agar (PCA) (log cfu/gram) during fermentation of jambal roti The total number of bacteria decreased (1 log cycle) during the fermentation process.
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