In the present study, conditions for Bacillus mucilaginous fermentation using Agaricus bisporus wastewater as culture medium were optimized. We analyzed the total number of living B. mucilaginous in the fermentation broth using multispectral imaging flow cytometry. Single-factor experiments were carried out, where a Plackett–Burman design was used to screen out three factors from the original six factors of processing wastewater solubility, initial pH, inoculum size, liquid volume, culture temperature, and rotation speed that affected the total number of viable B. mucilaginous. The Box–Behnken response surface method was used to optimize interactions between the three main factors and predict optimal fermentation conditions. Factors significantly affecting the total number of viable B. mucilaginous, including shaking speed, culturing temperature, and initial pH, were investigated. The optimum conditions for B. mucilaginous fermentation in A. bisporus wastewater were a rotational speed of 195 rpm, culture temperature of 29 °C, initial pH of 6.5, solubility of 0.5%, 8% inoculation volume, and 90 mL liquid volume in a 250 mL flask, culture time of 48 h. Under these conditions, the concentration of total viable bacteria reached 2.16 ± 0.02 × 108 Obj/mL, which meets the national standard. A. bisporus wastewater can be used for the cultivation of B. mucilaginous.

/pdf/optimization-of-the-culture-condition-of-bacillus-22vx4xe27r.pdf

Optimization of the culture condition of Bacillus mucilaginous using Agaricus bisporus industrial wastewater by Plackett–Burman combined with Box–Behnken response surface method

(1) Objective: The objective of this study was to screen amoxicillin (AMX)-degrading bacterial strains in pig manure and optimize the fermentation conditions for these strains to achieve high fermentation rate, which can provide an effective way for the practical application of bacterial strains as antibiotic-degrading bacterial in treating livestock waste for antibiotic residues. (2) Methods: Antibiotic susceptibility tests and high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) were employed to screen AMX-degrading bacterial strains in pig manure. The culture conditions were optimized for AMX-degrading bacterial strains using Plackeet-Burman design (PBD), the steepest ascent design, and the response surface methods, coupled with the Box-Behnken design (BBD). The effects of culture time, temperature, rotator (mixing) speed, inoculum level, and initial pH value on the growth of AMX-degrading strains were investigated. Experimental data obtained from BBD were utilized to generate a second-order polynomial regression model for evaluating the effects of the tested variables on the optical density at 600 nm (OD600) of culture solutions as the growth indicator for the screened AMX-degrading strains. (3) Results: The initial pH, culture time, and the inoculum level had significant effects on the OD600 value (growth) of the screened AMX-degrading strains. The initial pH value was found to be the most critical factor influencing the growth of bacteria. The optimized culture condition for the bacterial growth determined by the response surface methodology was: the initial pH of 6.9, culture time of 52 h, and inoculum level of 2%. The average OD value of 12 different fermentation conditions in the initial fermentation tests in this study was 1.72 and the optimization resulted in an OD value of 3.00. The verification experiment resulted in an OD value of 2.94, which confirmed the adequacy of the optimization model for the determining the optimal culture condition. (4) Conclusions: The growth of the screened strain of AMX-degrading bacteria could be optimized by changing the fermentation conditions. The optimization could be achieved by using the Box-Behnken response surface method and Plackett-Burman experimental design.

https://www.mdpi.com/1660-4601/17/6/1973/pdf

Optimization of Culture Conditions for Amoxicillin Degrading Bacteria Screened from Pig Manure.

The main attempt of this study is to isolate, determine potential probiotic properties and enzyme production of some lactic acid bacteria (LAB). Among all isolates, two LAB strains isolated from human mother milk and cottage cheese revealed antimicrobial activity against some tested pathogenic strains. Both isolates inhibited all the tested pathogens except Escherichia coli. The two isolates were identified by morphological, biochemical properties and then by 16S rRNA gene sequencing technique as Lactobacillus acidophilus SAM1 and Lactiplantibacillus plantarum SAM2. Potential probiotic characters were investigated. Both strains survived in relatively low pH and high bile concentrations and were able to grow at 0.5% of pancreatin concentrations. Their growth decreased by increasing phenol from 0.2% till 0.5%. Both strains did not show hemolytic activity. Coaggregation potential was exhibited by the two strains against Staphylococcus aureus and Candida albicans. Hydrophobicity of Lactobacillus acidophilus SAM1 and Lactiplantibacillus plantarum SAM2, with ethyl acetate; were 88.1% and 82.8%, respectively. Lactobacillus acidophilus SAM1 was susceptible to Ampicillin, Penicillin, Erythromycin, Ciprofloxacin and Tetracycline; on the contrary, it resists Vancomycin and Cefoxitin; while Lactiplantibacillus plantarum SAM2 resists all tested antibiotics. Maximum growth was achieved using glucose as a carbon source and yeast extract as nitrogen source for both strains; however, glucose is the most preferred carbon source for microorganisms and it prevents the uptake of carbon from other sources like yeast by catabolite repression mechanism. Lactobacillus acidophilus SAM1 produces lipase enzyme, while Lactiplantibacillus plantarum SAM2 produces amylase and protease. 

/pdf/identification-of-lactobacillus-strains-from-human-mother-3tjnejh0.pdf

Identification of Lactobacillus strains from human mother milk and cottage cheese revealed potential probiotic properties with enzymatic activity

The aim of this study was to optimize the cultural conditions for Bacillus megaterium using Agaricus bisporus industrial wastewater as nature culture through response surface methodology. In our present study, we analyzed the total number of living B. megaterium in the fermentation broth using multispectral imaging flow cytometry. Plackett-Burman design was applied to evaluate the effects of six variables, namely, initial pH, industrial wastewater solubility, rotating speed, culture temperature, inoculum size, and loading volume. Loading volume, initial pH, and culture temperature were found to influence the biomass of B. megaterium significantly and were further optimized by Box-Behnken design. After verification test, the optimum fermentation conditions of B. megaterium using the A. bisporus processing wastewater as nature culture media were obtained as follows: initial pH of 7.4, culture temperature of 25°C, loading volume of 40 mL/250 mL, culture time of 24 h, industrial wastewater solubility of 1%, rotating speed of 200 rpm, and inoculum size of 8%. The predicted optimum model's value was 8.88 × 108 Obj/mL and the average experimental value was 9.03 ± 0.02 × 108 Obj/mL, which met the national microbial fertilizers' standard. Furthermore, the field experiment results showed that the fermentation broth of B. megaterium could significantly improve the yield of Spinacia oleracea L.

/pdf/optimization-of-cultural-conditions-for-bacillus-megaterium-2pofvytv08.pdf

Optimization of Cultural Conditions for Bacillus megaterium Cultured in Agaricus bisporus Industrial Wastewater

Bioprospecting of probiotic lactic acid bacteria for cholesterol lowering and exopolysaccharide producing potential

The invention provides a protective agent in the freeze drying process of Lactobacillus acidophilus, which consists of 70 to 120g/L of defatted milk powder, 40 to 80mL/L of glycerol, 30 to 70g/L of trehalose, 05 to 2g/L of ascorbic acid, 3 to 85g/L of sodium chloride and 0 to 5g/L of gelatin The protective agent can make the survival rate of the Lactobacillus acidophilus subjected to freeze drying reach 983 percent, which is improved by 468 times compared with that (21 percent) of the Lactobacillus acidophilus subjected to the freeze drying without the protective agent; and the bacterium powder activity can reach 1*10 cfu/g

Protective agent in the freeze drying process of Lactobacillus acidophilus

The invention discloses a method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum. The method comprises the following steps of: adding lactobacillus acidophilus into a culture medium for culturing the lactobacillus acidophilus; after centrifugal collection of thalli, washing the thalli with physiological saline and centrifuging to remove clear liquid; after preparing the thalli into bacterial suspension by using a protecting agent I; adding a protecting agent II; and finally performing freeze-drying, wherein the formula of the protecting agent I comprises the following components: 10 to 20g/L trehalose, 10 to 20 g/L glycerol, 5 to 15 g/L bovine serum albumin, 6.5 to 8.5g/L sodium chloride and 60 to 100 g/L sucrose; and the formula of the protecting agent II comprises the following components: 150 to 250g/L skim milk powder, 10 to 40g/L glucose, 6.5 to 8.5g/L sodium chloride, and 0.2 to 0.3g/L ascorbic acid. The survival rate of the lactobacillus acidophilus powder prepared by the invention is up to 99.9 percent.

Method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum

The invention provides a lactobacillus acidophilus composite antifreeze factor and a freeze-drying method. According to a formula, 1 liter of the composite antifreeze factor comprises 2 to 6mL of glycerine, 2 to 8g of sodium chloride, 0 to 4g of trehalose and 0 to 0.8g of alanine. The method comprises the following steps of: adding the composite antifreeze factor into a culture medium for culturing the lactobacillus acidophilus; centrifuging and then collecting the lactobacillus acidophilus, cleaning the lactobacillus acidophilus by using normal saline, centrifuging and then removing clear liquid; and preparing the lactobacillus acidophilus into lactobacillus acidophilus suspension by using the normal saline, then uniformly mixing the lactobacillus acidophilus suspension with a protective agent and finally performing freeze drying. The freeze-drying antifreeze factor of the invention is directly added into the culture medium to prepare lactobacillus acidophilus powder; after freeze drying, the maximum survival rate of the lactobacillus acidophilus reaches 82.75 percent and a powder activity reaches over 1*10 cfu/g; and the survival rate of the lactobacillus acidophilus is over three times the survival rate (40.67 percent) of the lactobacillus acidophilus powder prepared without the antifreeze factor for the freeze drying.

Lactobacillus acidophilus composite antifreeze factor and freeze-drying method

The invention provides a multifunctional rotary evaporating device, comprising a bracket, first condensing equipment and second condensing equipment, an extracting device and a pipeline, a connecting channel, an evaporator and heating equipment, wherein the first condensing equipment and the second condensing equipment are fixedly connected with the bracket; the extracting device and the pipeline are respectively communicated with the first condensing equipment and the second condensing equipment; the connecting channel is communicated with the extracting device and the pipeline simultaneously; the evaporator is communicated with the connecting channel; the heating equipment heats the evaporator; the connecting channel is provided with a first rotating piston at the connecting position with the extracting device; and the connecting channel is provided with a second rotating piston at the connecting position with the pipeline. Therefore, the multifunctional rotary evaporating device not only can carry out extraction, but also can carry out backflow and separation, saves cost and has simple operation.

Multifunctional rotary evaporating device

The invention discloses a composite anti-freezing factor for lactobacillus acidophilus. Each liter of composite anti-freezing factor comprises 0.1 to 0.2 g of inositol, 3 to 4 g of sodium chloride, 0 to 0.07 g of bovine serum albumin and 0.03 to 0.06 g of alanine. The maximum survival rate of lactobacillus acidophilus powder prepared by adding the composite anti-freezing factor into a culture medium is 76.5 percent, while the data is only 52.7 percent when the composite anti-freezing factor is not added.

Composite anti-freezing factor for lactobacillus acidophilus

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