Fish sauce fermentation using Marinococcus halotolerans SPQ isolate as a starter culture.

A total of 344 halophilic bacteria were isolated from fish fermentation broths, solar salt crystals, seawater, and muds from ponds of salt pans in Vietnam and subjected to aroma evaluation using fish broth containing 29 ~ 30% (w/v) NaCl. One isolate from a salt crystal with the highest aroma score was selected, identified by using 16S rDNA sequence, and named Marinococcus halotolerans SPQ. The GC-MS results of the fish broth fermented by M. halotolerans SPQ revealed elevated concentrations of several aroma compounds such as ethyl alcohol, 1-propanol, 1-butyl alcohol, 1-amyl alcohol, and methionol. During the validation tests for M. halotolerans SPQ, using 2 kg of anchovy fish in 30% (w/v) NaCl at pH 5.78, the total and amino nitrogen values in the broth increased over time from 15.2 g/L at the beginning to 26.3 g/L at 6th month, with these values being comparable to those of the control. The ammoniacal nitrogen value (2.52 g/L) in the inoculated broth at 6th month was slightly higher than that (2.21 g/L) of control. The histamine content of the fish broth inoculated with M. halotolerans SPQ after 6 months was 110.12 mg/L, less than the maximum permitted safety limit of 200 mg/L, indicating it to be safe. Physical parameters, such as the total, amino, ammoniacal nitrogens, and histamine content of fish broth fermented by M. halotolerans MPQ met the standards for Vietnamese fish sauces. Two important umami amino acids, aspartic and glutamic acid, were seen to significantly increase, by 23.5% and 35.1%, respectively, even in the extremely harsh fermentation conditions posed by 30% (w/v) NaCl. The color, odor, and taste of the fish sauce fermented by M. halotolerans SPQ elicited the highest preference score accorded by the panelists. Taken together, M. halotolerans SPQ is a promising starter culture strain for fish sauce fermentation.

Screening and selection of bacteria inhibiting white spot syndrome virus infection to Litopenaeus vannamei.

A total of 173 bacterial strains were isolated from different sources at different regions such as fermented foods, shrimp guts, sea water, mangrove water, and sediments. These bacteria were screened against white spot syndrome virus (WSSV) infection in Palaemon paucidens. Based on mortality, white spot level, and healthiness, three bacterial strains were selected and identified using 16S rRNA gene sequencing. These bacterial strains were Bacillus subtilis KA1, B. licheniformis KA2, and B. subtilis KA3. WSSV challenge test in pilot scale was conducted using Litopenaeus vannamei with B. subtilis KA1 and B. subtilis KA3. The survival ratio of shrimp was 0% for WSSV control after 17th days, 84% for B. subtilis KA1 plus WSSV after 26th days, and 28% for B. subtilis KA3 with WSSV after 26th days. B. subtilis KA1 showed good growth at 18-37 °C in with and without 3% NaCl, and therefore can be applied to aquaculture at low to high temperatures. B. subtilis KA1 produced protease and lipase which can increase digestion to shrimp; exhibited antibacterial activity against Vibrio parahaemolyticus; and significantly increased the survival of WSSV challenged shrimps.

Statistical optimization of culture medium to produce recombinant viral protein by Escherichia coli host for diagnostic kit to detect human immunodeficiency virus (HIV) infection.

The maximal production of recombinant HIV1 gp41 by E. coli was examined in optimal culture condition and medium compositions. The culture condition such as growth, initial medium pHs, IPTG concentrations, induction times, temperature (0.5 OD, 7.6, 0.75 mM, 4.6 h, 32 °C respectively), and yeast extract (7.51 g/l), tryptone (7.26 g/l), glucose (2.45 g/l), NaCl (20.40 g/l), betaine (10.41 mM) and ampicillin (71.23 µg/ml) was optimized using statistical experimental design and response surface method (RSM). One of the main popular methods to attain high cell density in fed-batch culture is by controlling the nutrient feeding, which is often necessary for high yield in protein (0.63-0.72 mg/l) and cell (1.7-2 g/l) of the desired product in four litter fermentations.

Strain Selection and Optimization of Mixed Culture Conditions for Lactobacillus pentosus K1-23 with Antibacterial Activity and Aureobasidium pullulans NRRL 58012 Producing Immune-Enhancing ß-Glucan.

Lactobacillus pentosus K1-23 was selected from among 25 lactic acid bacterial strains owing to its high inhibitory activity against several pathogenic bacteria, including Escherichia coli, Salmonella typhimurium, S. gallinarum, Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium perfringens, and Listeria monocytogenes. Additionally, among 13 strains of Aureobasidium spp., A. pullulans NRRL 58012 was shown to produce the highest amount of ß-glucan (15.45 ± 0.07%) and was selected. Next, the optimal conditions for a solid-phase mixed culture with these two different microorganisms (one bacterium and one yeast) were determined. The optimal inoculum sizes for L. pentosus and A. pullulans were 1% and 5%, respectively. The appropriate inoculation time for L. pentosus K1-23 was 3 days after the inoculation of A. pullulans to initiate fermentation. The addition of 0.5% corn steep powder and 0.1% FeSO4 to the basal medium resulted in the increased production of lactic acid bacterial cells and ß-glucan. The following optimal conditions for solid-phase mixed culture were also statistically determined by using the response surface method: 37.84°C, pH 5.25, moisture content of 60.82%, and culture time of 6.08 days for L. pentosus; and 24.11°C, pH 5.65, moisture content of 60.08%, and culture time of 5.71 days for A. pullulans. Using the predicted optimal conditions, the experimental production values of L. pentosus cells and ß-glucan were 3.15 ± 0.10 × 108 CFU/g and 13.41 ± 0.04%, respectively. This mixed culture may function as a highly efficient antibiotic substitute based on the combined action of its anti-pathogenic bacterial and immune-enhancing activities.