Addition Of Olive Oil Research Articles

Effect of olive oil on the preparation of nanoemulsions and its effect on aroma release.

The present study focused on the effect of olive oil on Ostwald ripening of flavor nanoemulsions. The release of the aroma compounds from the nanoemulsion system was also investigated. The results showed that the droplets size of the nanoemulsions decreased sharply first and then kept stable with the increase of Tween 80. The optimum surfactant/cosurfactant (Km) ratio was determined at 7:1. The average particle size of nanoemulsion was 39.22nm. The polydispersity index (PDI) was 0.242nm, and the particle size distribution was in the range of 20-150nm at the optimum Km. The stability of the nanoemulsions was improved after the addition of olive oil, and it increased noticeably with the increase of olive oil. The addition of olive oil could help to stabilize the emulsions and hamper Ostwald ripening. All the 11 aroma compounds in the nanoemulsions were detected after 24-h storage. While only 5 aroma compounds were found after 48-h storage, and α-pinene and β-myrcene were the only two aroma compounds detected after 72-h storage with low contents of 1.41 and 0.5mg/L. The addition of olive oil inhibited the release of the aroma compounds from the nanoemulsion system. The released ethyl acetate was reduced by 48% after the addition of 10% olive oil. Significant decrease on the release of α-pinene and nonanal was observed after the addition of 3% olive oil. And the decrease was also observed on the release of β-myrcene, D-limonene, α-terpineol, decanal and eugenol when the olive oil content was ≥ 5%. However, benzyl alcohol, β-ionone and 1-octanol showed no significant changes with the increase of olive oil. This indicated that the addition of olive oil could provide greater retention of the aroma compounds in the nanoemulsions.

Manchette-acrosome disorders and testicular efficiency decline observed in hypercholesterolemic rabbits are recovered with olive oil enriched diet

High-fat diet is associated with hypercholesterolemia and seminal alterations in White New Zealand rabbits. We have previously reported disorders in the development of the manchette-acrosome complex during spermiogenesis and decreased testicular efficiency in hypercholesterolemic rabbits. On the other hand, olive oil incorporated into the diet improves cholesterolemia and semen parameters affected in hypercholesterolemic rabbits. In this paper, we report the recovery—with the addition of olive oil to diet—from the sub-cellular mechanisms involved in the shaping of the sperm cell and testicular efficiency altered in hypercholesterolemic rabbits. Using morphological (structural, ultra-structural and immuno-fluorescence techniques) and cell biology techniques, a reorganization of the manchette and related structures was observed when olive oil was added to the high-fat diet. Specifically, actin filaments, microtubules and lipid rafts—abnormally distributed in hypercholesterolemic rabbits—were recovered with dietary olive oil supplementation. The causes of the decline in sperm count were studied in the previous report and here in more detail. These were attributed to the decrease in the efficiency index and also to the increase in the apoptotic percentage in testis from animals under the high-fat diet. Surprisingly, the addition of olive oil to the diet avoided the sub-cellular, efficiency and apoptosis changes observed in hypercholesterolemic rabbits. This paper reports the positive effects of the olive oil addition to the diet in the recovery of testicular efficiency and normal sperm shaping, mechanisms altered by hypercholesterolemia.

The role of phenolic compounds on olive oil aroma release

In this study, atmospheric pressure chemical ionisation-mass spectrometry (APCI-MS) was successfully applied to understand the effect of phenolic compounds on the release of olive oil aroma compounds. Eight aroma compounds were monitored under in-vivo and in-vitro dynamic conditions in olive oil with and without the addition of virgin olive oil (VOO) biophenols. Three model olive oils (MOOs) were set up with identical volatile compounds concentrations using a refined olive oil (ROO). Phenolics were extracted from VOOs and were added to two MOOs in order to obtain two different concentrations of phenolic compounds (P+ = 354 mg kg−1; P++ = 593 mg kg−1). Another MOO was without VOO biophenols (P−). Phenolic compounds impacted both the intensity and time of aroma release. In the in-vivo study, 1-penten-3-one, trans-2-hexenal and esters had lower release in the presence of higher levels of biophenols after swallowing. In contrast, linalool and 1-hexanol had a greater release. The more hydrophobic compounds had a longer persistence in the breath than the hydrophilic compounds. VOO phenolics-proline-rich proteins complexes could explain the binding of aroma compounds and consequently their decrease during analysis and during organoleptic assessment of olive oil.

Enhancement of carotenoids and lipids production by oleaginous red yeast Sporidiobolus pararoseus KM281507

ABSTRACTBioconversion of biodiesel-derived crude glycerol into carotenoids and lipids was investigated by a microbial conversion of an oleaginous red yeast Sporidiobolus pararoseus KM281507. The methanol content in crude glycerol (0.5%, w/v) did not show a significant effect on biomass production by strain KM281507. However, demethanolized crude glycerol significantly supported the production of biomass (8.64 ± 0.13 g/L), lipids (2.92 ± 0.03 g/L), β-carotene (15.76 ± 0.85 mg/L), and total carotenoids (33.67 ± 1.28 mg/L). The optimal conditions suggested by central composite design were crude glycerol concentration (55.04 g/L), initial pH of medium (pH 5.63) and cultivation temperature (24.01°C). Under these conditions, the production of biomass, lipids, β-carotene, and total carotenoids were elevated up to 8.83 ± 0.05, 4.00 ± 0.06 g/L, 27.41 ± 0.20, and 53.70 ± 0.48 mg/L, respectively. Moreover, an addition of olive oil (0.5 − 2.0%) dramatically increased the production of biomass (14.47 ± 0.15 g/L), lipids (6.40 ± 0.09 g/L), β-carotene (54.43 ± 0.95 mg/L), and total carotenoids (70.92 ± 0.51 mg/L). The oleic acid content in lipids was also increased to 75.1% (w/w) of total fatty acids, indicating a good potential to be an alternative biodiesel feedstock. Meanwhile, the β-carotene content in total carotenoids was increased to 76.7% (w/w). Hence, strain KM281507 could be a good potential source of renewable biodiesel feedstock and natural carotenoids.

Efficient Production of Acid-Form Sophorolipids from Waste Glycerol and Fatty Acid Methyl Esters by <i>Candida floricola</i>

We discovered that Candida floricola ZM1502 is capable of selectively producing the promising hydrophilic biosurfactants, acid-form sophorolipids (SLs), from glycerol. However, productivity was very low (approximately 3.5 g L-1) under the initial culture conditions. Here, we describe the design of culture medium for abundant production of acid-form SLs by C. floricola ZM1502 using waste glycerol and hydrophobic substrates in order to develop a method for SL production and disposal of waste glycerol produced by oleo-chemical industries. Urea provided the best nitrogen source for acid-form SL production from glycerol among four nitrogen sources tested [urea, NaNO3, NH4NO3, and (NH4)2SO4]. Among carbon sources we compared, hydrophobic substrates (soybean oil and oleic acid) led to productivities of approximately 20 g L-1, indicating that hydrophobic substrates provided fatty acid moieties for SL production. Addition of olive oil and oleic acid to waste glycerol enhanced acid-form SL production to 42.1 ± 0.9 and 37.5 ± 3.4 g L-1, respectively. To develop a potential industrial process, we explored other suitable hydrophobic substrates for SL production, which were obtained on site from oleo-chemical industries. Alkyl C18 esters (Pastell M-182), along with waste glycerol, increased acid-form SL production to 48.0 ± 3.4 g L-1 over a 7-d period. Furthermore, we demonstrated abundant production of acidic SLs at the mini-jar fermenter scale, obtaining 169 g L-1 over 180 h using a fed-batch cultivation technique. Efficient acid-form SL production by C. floricola could have a great impact on the development of bio-industrial processes using waste glycerol as a substrate.

Effects of pomegranate seed oil and fermented juice polyphenols fraction in different solvents on copper-induced LDL oxidation

ABSTRACTEffects of pomegranate seed oils extracted with supercritical fluid or cold press methods (SFE-PSO and CP-PSO), unsaponifiable (N), and fermented juice polyphenols (W) fractions of pomegranate as well as olive oil and alpha-tocopherol, as pure or in different solvents were evaluated on preventing copper-induced LDL oxidation. The results showed addition of pure SFE-PSO, CP-PSO, N fractions, and olive oil had no significant effect on LDL oxidation. Also using various solvents did not help in the term of extending the oxidation lag phase. However, adding 0.5, 1, 1.5, and 3 µL of the W solution in EtOH could delay the lag phase 103.5, 145.9, 200, and 199.1 min, respectively. There was no synergistic effect between W and N on inhibiting the oxidation. It can be concluded that polyphenolic fraction of pomegranate juice can act as antiatherogenic supplementation and natural preservative for meat and fatty foods through direct inhibition of LDL oxidation.

Proximate Composition and Sensory Characteristics of Traditional and Oven-Drying Smoked Tilapia Fillets Enriched With Olive Oil

The study aimed to investigate the chemical and organoleptic characteristics of smoked tilapia fish fillets which were produced using different drying process and enriched with olive oil. Smoked fish were made using the following method: the fish were fasted overnight, shocked with cold water (4-5 oC) and then slaughtered and filleted. The fillets were soaked for 30 minutes in a brine solution containing spices and 5% (v/v) of liquid smoke. 5% (v/v) of olive oil was added into the solution and compared with control without olive oil addition. After 30 minutes, fish were cleaned from the spices using water and smoked at a temperature of 80 oC for 4-5 hours. The drying process was conducted in two different methods, i.e using oven-drying and traditional smoking. The smoked tilapia products were analyzed for proximate composition and sensory profiles. The result showed that addition of olive oil and methods of drying affected the proximate composition as well as sensory characteristics of the tilapia fillets. The fillets with addition of 5% olive oil and processed with oven-drying were considered as the best treatment due to low moisture content but high ash, fat and protein contents with 45.6±0.4%; 6.8±0.1%; 7.9±0.1% and 82±1.6%, respectively. Based on panelist evaluation, the fillets with 5% olive oil and processed with oven-drying also had high performances in appearance and texture.

Protection by extra virgin olive oil against oxidative stress in vitro and in vivo. Chemical and biological studies on the health benefits due to a major component of the Mediterranean diet.

We report the results of in vivo studies in Caenorhabditis elegans nematodes in which addition of extra virgin olive oil (EVOO) to their diet significantly increased their life span with respect to the control group. Furthermore, when nematodes were exposed to the pesticide paraquat, they started to die after two days, but after the addition of EVOO to their diet, both survival percentage and lifespans of paraquat-exposed nematodes increased. Since paraquat is associated with superoxide radical production, a test for scavenging this radical was performed using cyclovoltammetry and the EVOO efficiently scavenged the superoxide. Thus, a linear correlation (y = -0.0838x +19.73, regression factor = 0.99348) was observed for superoxide presence (y) in the voltaic cell as a function of aliquot (x) additions of EVOO, 10 μL each. The originally generated supoeroxide was approximately halved after 10 aliquots (100 μL total). The superoxide scavenging ability was analyzed, theoretically, using Density Functional Theory for tyrosol and hydroxytyrosol, two components of EVOO and was also confirmed experimentally for the galvinoxyl radical, using Electron Paramagnetic Resonance (EPR) spectroscopy. The galvinoxyl signal disappeared after adding 1 μL of EVOO to the EPR cell in 10 minutes. In addition, EVOO significantly decreased the proliferation of human leukemic THP-1 cells, while it kept the proliferation at about normal levels in rat L6 myoblasts, a non-tumoral skeletal muscle cell line. The protection due to EVOO was also assessed in L6 cells and THP-1 exposed to the radical generator cumene hydroperoxide, in which cell viability was reduced. Also in this case the oxidative stress was ameliorated by EVOO, in line with results obtained with tetrazolium dye reduction assays, cell cycle analysis and reactive oxygen species measurements. We ascribe these beneficial effects to EVOO antioxidant properties and our results are in agreement with a clear health benefit of EVOO use in the Mediterranean diet.

Low pressure supercritical CO2 extraction of astaxanthin from Haematococcus pluvialis demonstrated on a microfluidic chip

This study demonstrates the efficacy of low pressure supercritical CO2 extraction of astaxanthin from disrupted Haematococcus pluvialis. A microfluidic reactor was employed that enabled excellent control and allowed direct monitoring of the whole process at the single cell level, in real time. Astaxanthin extraction using ScCO2 achieved 92% recovery at 55 °C and 8 MPa applied over 15 h. With the addition of co-solvents, ethanol and olive oil, the extraction rates in both experiments were significantly improved reaching full recovery within a few minutes. Notably, for the ethanol case, the timescales of extraction process are reduced 1800-fold from 15 h to 30 s at 55 °C and 8 MPa, representing the fastest complete astaxanthin extraction at such low pressures.

Thermal degradation assessment of canola and olive oil using ultra-fast gas chromatography coupled with chemometrics

Oil blending is often used to enhance the properties of vegetable oils. The admixture of a more thermally stable oil makes the resulting blend more suitable for use in frying. A new method of quality assessment of vegetable oils used in frying is presented in this paper. In this method, ultra-fast gas chromatography coupled with flame ionization detector and chemometrics is employed. Principal component analysis was used for data processing. The results obtained with this method were compared with the results of the Rancimat test and sensory evaluation. It is demonstrated that the addition of olive oil improves the stability of rapeseed oil, and also changes its flavour and aroma profile. In addition, it was found that ultra-fast GC coupled with chemometrics is an effective tool for the assessment of the quality of edible oils. The proposed method does not require sample preparation, and the total time of analysis is less than 2 min.Graphical abstract

Purification and Characterization of Lipase from <i>Aspergillus flavus</i> PW2961 using Magnetic Nanoparticles.

Lipase from Aspergillus flavus was purified in a single step purification using MnFeO4 magnetic nano particles to achieve a 20.53- fold purification with specific activity of 11.29 U/mg and a 59% recovery yield. SDS-PAGE of lipase showed a single pure band with corresponding molecular weight of 35 kDa. The optimal temperature and pH for the enzyme activity were 45°C and 7.0 respectively. Addition of olive oil (1 %w/v) enhanced pH stability of the lipase with 86% residual activity at pH 7.0 after 6 h of incubation while the lipase was thermostable with 79% residual activity after 4 h of incubation. The enzyme activity was enhanced by Ca 2+ , Mg 2+ and Mn2+ while the presence of Cu 2+ , Co 2+ and Zn 2+ exhibited inhibitory effect on the enzyme. The study therefore presented lipase from Aspergillus flavus PW2961 with potential in industrial and biotechnological application. Key words : Lipase, purification, Aspergillus flavus PW2961, magnetic nanoparticles

The E/Z isomer ratio of lycopene in foods and effect of heating with edible oils and fats on isomerization of (all‐E)‐lycopene

Since Z‐isomers of lycopene are more bioavailable and show a higher antioxidant capacity than the (all‐E)‐isomer, it is important to investigate foods containing high amount of the Z‐isomer and develop practically feasible method for Z‐isomerization of (all‐E)‐lycopene. First, we investigated the E/Z isomer ratio of lycopene in raw and commercially available processed tomato products using an improved normal‐phase HPLC method. The tomato products contained 4.6–33.4% of Z‐isomers to the total lycopene, (5Z)‐lycopene being the most abundant Z‐isomer. The oil‐containing products like tomato sauce and tomato soup suffered heat processing contained a higher percentage of Z‐isomers of lycopene (27.4–33.4%). Subsequently, the impact of the amount and types of oils added on thermal Z‐isomerization of (all‐E)‐lycopene contained in tomato puree was investigated. Increased addition of olive oil to tomato puree increased the production of lycopene Z‐isomers upon heating at 120°C. (all‐E)‐Lycopene contained in tomato puree was converted to Z‐isomers in the range of 39.2–50.7%, when 5% of vegetable oil (linseed, soybean, corn, sesame, rapeseed, rice bran, safflower seed, olive, sunflower seed, or coconut oils) or animal fat (beef tallow and pork lard) was added before heating at 120°C for 30 min. When sesame oil was employed, the total Z‐isomerization ratio and (5Z)‐lycopene content were significantly increased.Practical applications: The dietary intake of lycopene offers many health benefits such as decreased risk of cancer and arteriosclerosis. Lycopene has a large number of geometric isomers caused by E/Z isomerization at arbitrary sites within the eleven conjugated double bonds, and the functionalities such as antioxidant capacity and bioavailability of the Z‐isomers are higher than those of the all‐E‐isomer. This study clarified the foods richly containing Z‐isomers of lycopene and demonstrated thermal Z‐isomerization of (all‐E)‐lycopene contained in tomato puree with edible oils and fats. These findings will contribute to effective intake of lycopene and the development of facile isomerization of (all‐E)‐lycopene to Z‐isomers in the fields of food, drink, and dietary supplement manufacturing, as well as for daily cooking at home.The Z‐isomers of lycopene contents in processed tomato products and thermal isomerization of (all‐E)‐lycopene to the Z‐isomers with edible oils and fats were investigated. The tomato products cooked with oils contained higher levels of the Z‐isomers, and sesame oil had excellent promoting effect of thermal Z‐isomerization of (all‐E)‐lycopene.

Effect of olive oil and freeze dried beetroot powder on quality characteristics of chicken meat patties by response surface methodology study

A two factor Central Composite Design (CCD) was applied to study the major effects and interactions of processing variables such as olive oil (5-15%) and beetroot powder (1-5%) on cooking properties of chicken patties. The addition of olive oil increased reduction in diameter, shrinkage and emulsion stability values however, reduction in thickness, fat and moisture retention, cooking yield values decreased as the amount of olive oil increased. On the other hand, the incorporation of beetroot powder decreased reduction in thickness and diameter, shrinkage and emulsion stability. Cooking yield and fat and moisture retention values were found to increase by the addition of beetroot powder.

Phytochemical composition and in vitro anti-tumour activities of selected tomato varieties.

Previous studies indicated that tomato is a rich source of phytochemicals that act on different tumours. In this research, the phytochemical composition of selected tomato varieties was assessed by GLC and UHPLC/HPLC-MS, as well as their anti-tumour activities on HT-29 colorectal cancer cells. Significant differences were found among tomato varieties; lycopene was high in Racimo, phenolics in Pera, sterols in Cherry, and linoleic acid predominated in all varieties. The MTT and LDH assays showed significant time- and concentration-dependent inhibitory/cytotoxic effects of all tomato varieties on HT-29 cells. Furthermore, the joint addition of tomato carotenoids and olive oil to HT-29 cell cultures induced inhibitory effects significantly higher than those obtained from each of them acting separately, while no actions were exercised in CCD-18 normal cells. Tomato fruits constitute a healthy source of phytochemicals, although differences exist among varieties. In vitro, all of them inhibit colorectal cancer cell proliferation with Racimo variety at the top, and exercising a selective action on cancer cells by considering the lack of effects on CCD-18 cells. Furthermore, synergy was observed between olive oil and tomato carotenoids in inhibiting HT-29 cancer cell proliferation; conversely, phenolics showed no significant effects and hindered carotenoids actions. © 2016 Society of Chemical Industry.

Effect of fat replacement by olive oil on the physico-chemical properties, fatty acids, cholesterol and tocopherol content of pâté

The present study investigates the effects of olive oil when used as back fat replacers on the physico-chemical properties, fatty acids, cholesterol and α-tocopherol content of p.t.. The replacement of back fat by olive oil at 50 and 100% did not affect the moisture, fat or protein contents, while it increased yellowness. According to texture parameters, reformulated p.t. presented lower values for hardness, cohesiveness, gumminess and chewiness. The use of olive oil significantly (P < 0.05) affected the fatty acid content. The amount of MUFA increased, while the content of SFA and PUFA decreased as the back fat was replaced by olive oil. The replacement of fat resulted in an improvement in all nutritional indexes. Atherogenic and thrombogenic indexes decreased while the h/H ratio increased with fat replacement. At the same time, the content of α-tocopherol increased and the amount of cholesterol decreased with the addition of olive oil. Therefore, olive oil provides p.t. with high levels of C18:1n9c and MUFA, natural antioxidants such as α-tocopherol and reduces cholesterol levels. As a general conclusion, the replacement of back fat by olive oil allows us to obtain a healthier product.

The Effect of Olive Oil Addition into Oatmeal in Lowering Total Blood Cholesterol and LDL (Low Density Lipoprotein) in Wistar Rat

Oat contains beta-glucan fiber and olive oil contains unsaturated fatty acids which havebeen proven to decrease blood cholesterol. This research compared the effect between acombination of oatmeal and olive oil and each food as single intake in decreasing total bloodcholesterol and LDL cholesterol. We used 25 rats (Rattus norvegicus L.), divided them into fivegroups; negative control (KN), positive control (KP), olive oil group (KMZ), oatmeal group(KBG), and combination group (KMZ+KBG). The total blood cholesterol and LDL cholesterollevels were measured after 14 days of induction and after 28 days of treatment. Data wereanalyzed using one way ANOVA, continued with LSD test with α = 0.05. The result showedthere were 10.85%, 8.85% and 6.60% lower in total blood cholesterol level using KMZ+KBG,KMZ and KBG respectively and 13.85%, 9.33% ,and 8.80% lower in LDL cholesterol levelusing KMZ+KBG, KMZ and KBG respectively. ANOVA test showed a significant decrease ineach treatment. LSD test showed that there was a significant difference between KMZ+KBGcompared to KBG in total blood cholesterol and LDL lowering, but did not showed significantdifference compared to KMZ.Keywords: oatmeal, olive oil, total blood cholesterol, LDL

Kkuaripepper (Capsicum annum L.) and Olive Oil Effects on Quality Characteristics of Pork Sausage Studied by Response Surface Methodology

Response surface methodology was used to investigate the effect and interactions of processing variables such as kkuaripepper powder (1.0-5.0%), olive oil (3-13%) on physicochemical, textural and sensory properties of cooked pork sausage. It was found that L* and a* values decreased; however, water-holding capacity, and a* values increased, respectively, as the amount of kkuaripepper increased. Olive oil addition decreased water-holding capacity, moisture contents, and L* values of the sausages. The adhesiveness depended on the kkuaripepper and olive oil added, as its linear effect was positive at p<0.01. Kkuaripepper addition increased the springiness and chewiness of sausages. The preference of color, flavor, taste, and overall quality depend on the addition of kkuaripepper and olive oil. The optimum formulation for pork sausage with kkuaripepper powder was 2.99 g of kkuari pepper powder and 9.12 g of olive oil. The results of this optimization study would be useful for meat industry that tends to increase the product yield for sausages using the optimum levels of ingredients by RSM (Response surface methodology).

Not Simply a Matter of Fish Intake.

Recent findings have highlighted enhanced fish consumption as a potential measure to increase intake of healthy fatty acids, particularly omega-3. The generalizability of this recommendation, however, may fall short of differences in fish species and cooking techniques. Hence, we investigated how these 2 variables affect the lipid content in fish flesh. Nine species of freshwater, deep sea or shore fish were grilled, steamed or fried with or without the addition of soybean oil, olive oil or butter. The lipid composition was analysed and a significant difference was observed in cholesterol, saturated fatty acids, polyunsaturated fatty acids, omega-3 fatty acids and omega-6 fatty acids contents between species (p<0.05). The use of soybean or olive oil was associated with a significant change in flesh concentration of polyunsaturated, omega-3 and omega-6 fatty acids (p<0.05). This study calls attention to the specific lipid content that must be expected from different fish species and cooking techniques.

Correction of Mitochondrial Enzyme Activities in the Skeletal Muscles of Old Rats in Response to Addition of Olive Oil to the Ration.

Activities of mitochondrial electron transport chain enzymes NADH-CoQ oxidoreductase (complex I), cytochrome C-oxidase (complex IV), and citrate synthase were measured by spectrophotometry in m. quadriceps femoris homogenate from old rats receiving olive oil with the ration. Reduced activities of complexes I and IV in old animals were restored to the level of young animals after 6-week consumption of olive oil. Activity of citrate synthase did not change with age. Positive effect of olive oil on fatty-acid composition of the muscle tissue in old animals was demonstrated. The content of summary monounsaturated fatty acids, reduced with aging, and of summary polyunsaturated ones, increasing with age, were restored in old rats to the levels virtually not differing from the levels of young animals.

Isolation and Identification Probiotic Candidates Bacteria from Crossbreed Fries Holland (CFH) Colostrum That Grown on Nutrient Agar with Olive Oil and Casein Enrichment

Aims of the research were to isolate the bovine colostrum indigenous bacteria that potent as probiotics with Nutrient Agar (NA) as a growth media. Selected isolate expected to develop as anti-pathogen in food spoilage and digestion. Material used including bovine colostrum taking from lactating Crossbreed Fries Holland (CFH) 1-3 days after partus which done compositely. Crossbreed Fries Holland colostrum everyday was taken 250 mL at the same time in the morning and afernoon and then stored at the refrigerator. Research done with qualitative method which analyzes descriptively through isolation step cover: instrument sterilization, dilution, morphology observation and pure culture production, then continued with identification step (gram staining, catalase test, oxidase test, low pH, high bile salt and API Test). Result showed that isolation with NA with olive oil addition obtained 10 bacteria (5 Bacillus genus, 3 Micrococcus genus and Staphylococcus genus). While at the NA with 1% casein addition obtained 12 pure isolate including: 2 Staphylococcus genus, 5 Bacillus genus, 2 Micrococcus genus and each Streptococcus, Enterobacter and Veillonella was found 1. Catalase and oxidase test resulting in two groups that cover Bacillus and Coccus group. Twenty isolate identified as gram positive bacteria, while two genus such as Enterobacter and Veillonella was identified as gram negative bacteria. Gram positive bacteria tested to determine the species using API test and resulting two dominant bacteria species with code Ic8 for Bacillus coagulans species with 93.7% similarity, Ic12 for Aneurinibacillus aneurinilyticus with 99.1% similarity.