SODIUM BENZOATE ASSIMILATION BY RHODOCOCCUS AETHERIVORANS UCM AC-602

Sodium benzoate (SB) is a widely used preservative (E211) and also serves as an active ingredient in several pharmaceuticals. Prolonged exposure to SB can disrupt aquatic ecosystems, adversely affecting aquatic organisms and potentially human health. Aim. To investigate the features of sodium benzoate biodegradation by the Rhodococcus aetherivorans UCM Ac-602 strain and assess the ecological safety of the toxicant and its degradation products for higher plants. Methods. The concentration of SB was determined using high-performance liquid chromatography. Fatty acids composition was analyzed by gas chromatography–mass spectrometry. Catalase activity was measured spectrophotometrically, and cell membrane permeability was assessed using crystal violet. Phytotoxicity was evaluated via a rapid assay using wheat (Triticum aestivum L.) as a test plant. Results. The R. aetherivorans UCM Ac-602 fully utilized 0.5 g/L of SB within 7 days. A twofold decrease in the C18:1 cis-9 fatty acid and a 1.7-fold increase in 10Me-C18:0 were observed during growth on SB. Changes in catalase activity and membrane permeability during SB assimilation contributed to cellular protection against the toxic effects of the substrate. Neither SB nor its metabolites exhibited phytotoxic properties. Conclusions. The main mechanisms of adaptation of R. aetherivorans UCM Ac-602 to SB assimilation are modifications in fatty acid profiles, changes in catalase activity, and alterations in membrane permeability. SB and its degradation products were shown to be non-phytotoxic and safe for plant development.

Integrin signaling and membrane blebbing modulate cell adhesion, spreading, and migration. However, the relationship between integrin signaling and membrane blebbing is unclear. Here, we show that an integrin-ligand interaction induces both membrane blebbing and changes in membrane permeability. Sodium-proton exchanger 1 (NHE1) and sodium-calcium exchanger 1 (NCX1) are membrane proteins located on the bleb membrane. Inhibition of NHE1 disrupts membrane blebbing and decreases changes in membrane permeability. However, inhibition of NCX1 enhances cell blebbing; cells become swollen because of NHE1 induced intracellular sodium accumulation. Our study found that NHE1 induced sodium influx is a driving force for membrane bleb growth, while sodium efflux (and calcium influx) induced by NCX1 in a reverse mode results in membrane bleb retraction. Together, these findings reveal a novel function for NHE1 and NCX1 in membrane blebbing and permeability, and establish a link between membrane blebbing and integrin signaling.

This study is to determine the shelf life of kunuzaki under different temperatures regimes, and the role of additives to the alteration in its shelf life or chemical components and also to possibly find a means of extending its shelf life using additives and temperature without the lowering its nutritional quality. During the study it was observed that appropriate concentrations of sodium benzoate and pasteurization can be used as preservative and preservative method respectively for extension of the shelf life of kununzaki under ambient temperature (30-31 0 C). Sodium benzoate, when used at a concentration of (sodium benzoate (0.144g) for 35cl of kununzaki, slows down undesirable changes in the physicochemical characteristics of kunun zaki. This concentration of sodium benzoate brought about the least changes in pH compared to the other preservative treatments and also exhibited the best potential for use in the preservation of kunun zaki. Sodium benzoate can be used to extend shelf life of kunun zaki up to a period of three (3) days when preserved at ambient temperature (30-31 0 C). In addition to having the ability to extend the shelf life of kunun zaki, sodium benzoate has no side effects to the consumer when put in appropriate quantity. DOI: 10.7176/JEES/12-3-02 Publication date: March 31 st 2022

Microfluidic colorimetric analysis system for sodium benzoate detection in foods

Effects of sodium benzoate on growth and physiological characteristics of wheat seedlings under compound heavy metal stress

This investigation was designed to evaluate the reproductive toxicity induced by potassium bromate (at doses of 50, 40, 30, 20, and 10 mg/kg), sodium benzoate (at doses of 5, 4, 3, 2, and 1 mg/kg), and caramel IV (at a dose of 200 mg/kg for 1, 2, 3, 4, and 5 weeks) in male albino mice through the assessments of sperm abnormality and sperm count. The concentrations of the three additives tested were chosen purposefully based on the acceptable daily intake of each of the additives by standard organizations. The level of Total Testosterone (TT), Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH), as well as the activities of catalase (CAT), superoxide dismutase (SOD), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), were also measured. The results obtained showed that all three tested food additives led to a statistically significant (p < 0.05) increase in abnormal sperm cells among exposed mice. The sperm abnormalities observed include folded sperm, amorphous heads, distal droplets, absent hooks, improper tail attachments, and looped tails. Furthermore, it was observed that only mice exposed to potassium bromate and caramel exhibited a statistically significant decrease in mean sperm count. Alterations in the levels and activities of TT, ALT, FSH, AST, LH, and ALP were also observed to be concentration-dependent in the exposed mice. The findings suggest that the reproductive toxicity and changes in enzyme activities in the treated mice are dependent on the concentration of potassium bromate and sodium benzoate, as well as the duration of exposure to caramel IV. These indicate that high consumption of foods or beverages containing potassium bromate, sodium benzoate, and caramel IV colourant may induce reproductive abnormalities in the consumers.

Seasonal and spatial shifts in copepod diets within tropical estuaries measured by fatty acid profiles

Cheddar-type cheese produced from buttermilk had softer texture than standard cheddar cheese due to lower fat content of buttermilk. Fat is extremely important for the functional characteristics and optimum textural attributes of cheese. The effect of different fat contents of buttermilk on chemical characteristics of cheddar-type cheese is not previously investigated. This investigation was conducted to know the effect of different fat contents of buttermilk on fatty acids composition, organic acids, vitamins, lipolysis and sensory characteristics of cheddar-type cheese. Cheddar-type cheese was produced from buttermilk having 1, 1.75, 2.50 and 3.25% fat contents (control, T1, T2 and T3). Fat content of control, T1, T2 and T3 were 9.81, 16.34, 25.17 and 31.19%. Fatty acids profile was determined on GC–MS, organic acids and vitamin A and E were determined on HPLC. Free fatty acids, peroxide value and cholesterol were determined. Cheddar-style cheese produced from buttermilk (1% fat) showed that it had softer texture and lacking typical cheese flavor. Gas chromatography–mass spectrometry (GC–MS) analysis showed that long-chain unsaturated fatty acids in control, T1, T2 and T3 samples were 45.88, 45.78, 45.90 and 46.19 mg/100 g. High Performance Liquid Chromatography (HPLC) analysis showed that lactic acid, propionic acid, citric acid and acetic acid gradually and steadily increased during the storage interval of 90 days. At the age of 90 days, lactic acid in control, T1, T2 and T3 was 4,789, 5,487, 6,571 and 8,049 ppm, respectively. At the end of ripening duration of 90 days, free fatty acids in control, T1, T2 and T3 were 0.29, 0.31, 0.35 and 0.42% with no difference in peroxide value. Stability of vitamin A after 90 days storage control, T1, T2 and T3 was 87.0, 80.0, 94.0 and 91.0%. Flavor score of cheddar-type cheese produced from butter milk having 1.0, 2.5 and 3.25% fat content was 81, 89 and 91% of total score (9). Hence, it is concluded that cheddar-type cheese can be produced from buttermilk having 2.5 and 3.25% fat contents with acceptable sensory attributes. Application of buttermilk for the production of other cheese varieties should be studied.

Sodium benzoates as a chemical preservative are common practice in modern food technology. It is permitted by international laws in food as a food additive with a restricted amount. But it is highly used in different food products like fruit juices. Therefore, an experimental study was conducted to determine the concentration of sodium benzoate in different brands of orange juices available in the market Tangail region, Bangladesh using High Performance Liquid Chromatography (HPLC). Chromatographic analysis was performed by a column (250 × 4.6 mm) with an isocratic solvent system at 40°C. The mobile phase consisted of sodium acetate and acetic acid buffer (pH was 4). The correlation of coefficient was 0.9999 from the standards curves and the variety of external standard absorptions was 0.93 to 20.05 µg/mL. Analysis of four brands of orange juice samples showed that the concentration of sodium benzoate was within the FDA standard, but two brand samples slightly exceeded the Bangladesh Standard and Testing Institute (BSTI) standard. Brand 1 and Brand 4 samples contained sodium benzoate 14.61 and 9.59 mg/100 mL respectively which were within the level BSTI standard range whereas Brand 2 and Brand 3 samples contained 16.11 and 16.33 mg/100 mL respectively which exceeded the level of BSTI standard.

Comprehensive biochemical analysis and nutritional evaluation of fatty acid and amino acid profiles in eight seahorse species (Hippocampus spp.)

Objective: Radiation-induced alterations in permeability of plasma membrane and associated apoptotic changes were investigated in apoptotic thymocytes induced by radiation. Materials and Methods: Immature mouse thymocytes suspended in RPMI 1640 were irradiated by γ-rays for desired doses up to 10 Gy and these cells were examined after incubation (up to 24 h) at 37°C. Radiation-induced changes in plasma membrane of thymocytes were determined by fluorescence technique using fluorescein diacetate (FDA) and trypan blue (TB) method. Induction of apoptosis in irradiated cells was determined using annexin-V as fluorescence probe method and measurement of cytosolic caspase-3 activity. Results: Radiation-induced apoptotic thymocytes showed an increase in membrane permeability as observed by leakage of FDA, which was poorly detected by TB. FDA could sensitively detect the dose-dependent variation in membrane permeability alterations in the range of 0.5–2 Gy incubated at 37°C. In radiation-induced apoptotic death, externalization of phosphatidylserine is an early event than caspase-3 activation. On incubation of cells, an increase in caspase-3 activity and proportion of annexin-positive cells was observed, which further increased after irradiation. Conclusion: Our results showed membrane permeability changes induced by γ-irradiation, which seem to be closely associated with the apoptosis in thymocytes.

A rapid, simple, and sensitive method is described for the determination of the food preservative sodium benzoate in soft drinks, various jams, and ketchup. The method utilizes high performance liquid chromatography followed by diode array detection. Chromatographic separation was achieved using a C18 reversed phase column and methanol: water (70∶30, v/v) adjusted to pH 3.45 with glacial acetic acid as mobile phase, 0.45 mL . min−1 flow rate, and UV detection at 245 nm. Amoxicillin was used as the internal standard. The retention time observed for sodium benzoate and amoxicillin were 5.01 and 12.07 min, respectively. The method is selective, reliable, and reproducible with a relative standard deviation of 0.66 and linear in the range of 50–450 ng/mL sodium benzoate concentration. The limit of detection and limit of quantification concentrations were 61 pg/mL and 203 pg/mL, respectively. The proposed method can be used for the routine analysis of sodium benzoate in soft drinks and jams.

Introduction. Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the world. Non-alcoholic steatohepatitis (NASH), a clinically progressive morphological form of NAFLD, ranks second in the list of reasons for liver transplantation in the adult population. In the pathogenesis of this disease, metabolism and distribution of free fatty acids (FFA) play an important role. A large number of studies have established that the level of FFA in peripheral blood directly correlates with the severity of NASH, but it is still unclear what effect fluctuations in the profile of fatty acids (FA) in the liver have in steatohepatitis.Aim. Study of changes in the profile of fatty acids in the liver of laboratory animals with experimental non-alcoholic steatohepatitis.Materials and methods. The study was carried out on 17 white outbred male rats, which were randomized into two groups – intact (n = 6) and control (steatohepatitis) (n = 11). Steatohepatitis was modeled by 12-month use of a hypercaloric high-fat diet against the background of hypodynamia. The content of fatty acids in the liver was determined in the reaction of methanolysis and extraction with a hexane mixture of their methyl esters. The LC was separated by gas chromatography-mass spectrometry. Calibration for quantitative calculation was carried out with deuterated tridecanoic acid. The content of saturated and monounsaturated higher FAs, their aldehydes and hydroxy derivatives, as well as sterols were studied.Results and discussion. A total decrease in the content of FFA in the liver of animals with steatohepatitis was revealed. The most significant decrease occurred mainly in the class of monounsaturated fatty acids and cholesterol. Also, a significant decrease in the activity of Δ9-desaturase, a key enzyme in the synthesis of monounsaturated FAs from their precursor with the same carbon chain length, was revealed, which was manifested by a significant decrease in their amount in the liver. There were no statistically significant changes in the levels of aldehydes and hydroxy acids between the study groups, as well as in the level of sterols (except for cholesterol, the content of which decreased significantly).Conclusion. Thus, in the liver of rats with steatohepatitis caused by a combination of a hypercaloric diet and hypodynamia, statistically significant changes in the profile and concentration of fatty acids were found in comparison with healthy animals. The demonstrated shifts in FA composition may reflect both adaptive and pathological changes in the liver of animals with NAFLD and require further study.

Dietary sodium benzoate improves growth, morphology, antioxidant capacity and resistance against Aeromonas hydrophila of Pacific white shrimp (Litopenaeus vannamei)

This study demonstrates a new electrochemical impedance spectroscopic (EIS) method for measurements of the changes in membrane permeability during the process of cell anoxia. Madin-Darby canine kidney (MDCK) cells were employed as the model cells and were cultured onto gelatin-modified glassy carbon (GC) electrodes. EIS measurements were conducted at the MDCK/gelatin-modified GC electrodes with Fe(CN)(6)(3-/4-) as the redox probe. The anoxia of the cells grown onto electrode surface was induced by the addition of carbonycyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) into the cell culture, in which the MDCK/gelatin-modified GC electrodes were immersed for different times. The EIS results show that the presence of FCCP in the cell culture clearly decreases the charge-transfer resistance of the Fe(CN)(6)(3-/4-) redox probe at the MDCK/gelatin-modified GC electrodes, and the charge-transfer resistance decreases with increasing time employed for immersing the MDCK/gelatin-modified GC electrodes into the cell culture containing FCCP. These results demonstrate that the EIS method could be used to monitor the changes in the cell membrane permeability during the FCCP-induced cell anoxia. To simulate the EIS system, a rational equivalent circuit was proposed and the values of ohmic resistance of the electrolyte, charge-transfer resistance and constant phase elements for both the gelatin and the cell layers are given with the fitting error in an acceptable value. This study actually offers a new and simple approach to measuring the dynamic process of cell death induced by anoxia through monitoring the changes in the cell membrane permeability.