Soheyl Reyhani Poul; Sakineh Yeganeh; Reza Safari
Abstract
[1]Introduction: Nisin is one of the antimicrobial substances that is used today as a preservative in various foodstuffs. It is a bacteriocin comprised of 34 amino acids and a molecular weight of 3.5 Da. With all the benefits of nisin, there are barriers to its use in dairy and protein rich products. ...
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[1]Introduction: Nisin is one of the antimicrobial substances that is used today as a preservative in various foodstuffs. It is a bacteriocin comprised of 34 amino acids and a molecular weight of 3.5 Da. With all the benefits of nisin, there are barriers to its use in dairy and protein rich products. One of these barriers is the combination of nisin with fats, proteins and sugars and the consequent reduction of its antibacterial activity. In the food science and industry, the use of the technique of encapsulation and production of liposome is the best possible solution in such cases. Also, by adding an antimicrobial agent such as chitosan to the coating of nanoliposomes, the antibacterial activity of the product may be increased. The aim of the present research was to produce nanoliposomes carrying nisin with (and without) chitosan coating and to evaluate the physical and antibacterial properties against two gram-positive bacteria, Bacillus cereus and Staphylococcus aureus. Materials and Methods: In this study, four treatments of nanoliposomes carrying nisin (NN), nanoliposomes carrying nisin coated with chitosan 0.05% ((NN-CH (0.05)), nanoliposomes carrying nisin coated with chitosan 0.1% (NN-CH (0.1)) and nanoliposomes carrying nisin coated with chitosan 0.5% (NN-CH (0.5)) were prepared and examined in terms of physical properties (average particle size, particle dispersity index, zeta potential and encapsulation efficiency) and antibacterial activity (against two gram-positive bacteria, Bacillus cereus and Staphylococcus aureus with two diffusion methods in agar medium and microdilution test). This research was conducted in a completely randomized design and SPSS and EXCEL softwares were used for statistical analysis and drawing of diagram, respectively. Data were analyzed by one-way analysis of variance and the difference between the means was evaluated by Duncan's test at 95% confidence level. Results and Discussion: The results showed that the average particle sizein different treatments with each other are significantly different (P<0.05) and vary from about 110 to 327nm; Also as the amount of chitosan in the coating increased, the particle size increased (P<0.05). This indicates the successful binding of chitosan to the surface of the nanoliposome, which results in the formation of a layer around the nanoliposome and an increase in particle size. Particle dispersity index was recorded less than 0.3 in all treatments and was not related to the amount of chitosan in the coating. With increasing the amount of chitosan in the coating of nanoliposomes, zeta potential increased significantly (P<0.05). This index changed from -55.34 in NN treatment to 53.14 mV in NN-CH (0.5) treatment. In fact, chitosan as a cationic polysaccharide changes the potential to positive values. As the amount of chitosan in coating of nanoliposomes increased, the encapsulation efficiency increased significantly in the treatments (P<0.05); this index increased from 32.19% in NN treatment to 75.14% in NN-CH (0.5) treatment. The results of the antibacterial activity of nisin in two methods of diffusion in agar medium and microdilution test showed that its antibacterial activity increased with nanoencapsulation of nisin with (and without) chitosan coating (p<0.05). Also, with the increase in chitosan concentration, the antibacterial activity of carrier nanoliposomes increased and the highest antibacterial activity was recorded in NN-CH (0.5) treatment (p<0.05). The diameter of the non-growth halo of Bacillus cereus against the research treatments (with five concentrations of 2.5 to 25 μg/ml) varied from about 4.5 to 17.5 mm. This amount for Staphylococcus aureus was recorded from 2.1 to 26.5 mm. By increasing the concentration of nisin and carrier nanoliposomes, the diameter of the halo of non-growth of both bacteria increased significantly (p<0.05). But an exception was recorded in this case; The diameter of the non-growth halo for Staphylococcus aureus in two concentrations of 2.5 and 5 μg/ml of treatments was the same and had no significant difference (p>0.05). The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the examined treatments for Bacillus cereus were in the range of 100 to 400 and 200 to 500 μg/ml, respectively. These two concentrations for Staphylococcus aureus were recorded as 50 to 200 and 100 to 400 μg/ml respectively. Based on the values of diameter of non-growth halo, MIC and MBC it can be claimed that Bacillus cereus is more resistant to the examined treatments than Staphylococcus aureus.Nanoencapsulation of nisin in the form of carrier nanoliposomes with chitosan coating is a suitable solution to improve its physical and antibacterial properties. In such a way that by increasing the concentration of chitosan in the coating, both of the aforementioned properties improved significantly. Nanoliposomes carrying nisin with (and without) chitosan coating have the ability to inhibit the growth and killing Bacillus cereus and Staphylococcus aureus bacteria. The antibacterial activity increases with the increase in nisin and carrier nanoliposomes concentrations. The value of non-growth halo, minimum inhibitory concentration and minimum bactericidal concentration confirm that Bacillus cereus is more resistant to nisin and its carrier nanoliposomes than Staphylococcus aureus.
Marzieh Omidi Mirzaei; Mohammad Hojjati; Behrooz Alizadeh Behbahani; Mohammad Noshad
Abstract
Introduction: Essential oils and secondary metabolites of plants have too many uses in medicine as well as food and hygiene industries. The herbal essential oils include different health features including antioxidant and antibacterial activities. Several forms of the activated oxygen, also known as ...
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Introduction: Essential oils and secondary metabolites of plants have too many uses in medicine as well as food and hygiene industries. The herbal essential oils include different health features including antioxidant and antibacterial activities. Several forms of the activated oxygen, also known as reactive oxygen species (ROS), include free radicals and non-free radical species. In traditional Iranian medicine, coriander seeds are widely used to treat the disease. The objectives of this paper were to identify the chemical compounds and to measure the phenol content and the antioxidant potential of coriander seed essential oil in addition to its free radical scavenging activity. The other aim of this work was to investigate the antimicrobial of coriander seed essential oil on Bacillus cereus, Salmonella typhi, Escherichia coli and Pseudomonas aeruginosa “in vitro”. Materials and methods: In this research, the coriander seed essential oil (100 g) was extraction using water-distillation method with clevenger apparatus. Afterwards, coriander seed essential oil was collected in vials which had already been weighed by a 0.0001 balance and stored at 4 °C until testing. Chemical composition of coriander seed essential oil was determined using gas chromatography. The antioxidant activity was determined by 2,2’-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) di-ammonium salt (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicles, respectively. The method of Folin-Ciocalteu was performed through determination of TPC. The result was reported as mg of gallic acid/g of the dried coriander seed essential oil. The antioxidant potential of the essential oil was compared with BHA synthetic antioxidant at a concentration of 100 μg/ml. Antibacterial activity of coriander essential oil was determined by disc diffusion agar (Kirby-Bauer test), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) methods. Results and discussion: Based on the results of chemical analysis, the coriander seed essential oil was rich in oxygenated monoterpenes (89.94%). The main compound of coriander seed essential oil was Linalool (76.75%). The highest percentage of free radical scavenging for DPPH was 53.5% and for ABTS 66.6% at 900 ppm concentration. The total phenol content essential oil was 38.04 ± 0.02 mg GAE/g. The result show that, the most sensitive and the most resistant bacteria with diameter inhibition zone 30.30 mm and 23.15 mm were Bacillus cereus and Salmonella typhi respectively. MIC of coriander seed essential oil for Bacillus cereus, Salmonella typhi, Escherichia coli and Pseudomonas aeruginosa was 2, 4, 4 and 4 mg/ml respectively. MBC of coriander seed essential oil for Bacillus cereus, Salmonella typhi, Escherichia coli and Pseudomonas aeruginosa was 512, > 512, > 512 and 512 mg/ml respectively. In general, the results indicated that the coriander seed essential oil was effective on microorganisms; nevertheless, the extent of its effectiveness varied depending on the type of microorganism. The gram-positive bacteria are more sensitive to essential oil rather than gram-negative ones. The higher resistance of gram-negative bacteria to the essential oils of medicinal plants could be attributed to the more complex structure of the cell membrane of these bacteria compared with the single layer structure of the gram-positive ones. The results of this study revealed that coriander seed essential oil had less antioxidant activity than synthetic antioxidant BHA. Antibacterial activity of the essential oil was higher than the gentamicin antibiotic. Regarding the chemical compositions identified in the coriander seed essential oil, these compositions could be employed as an important economical source uses in medicine as we as food and hygiene industries.
Samaneh Hizomi Shirejini; Hadi Koohsari; Seyedeh Zahra Seyed Alangi
Abstract
Introduction: Study in order to introduce new antimicrobial agents with natural origin to prevent the antibiotic resistance and eliminating its effects, employing chemical agents is an indispensable necessity. Honey, as an important food with natural origin has high antimicrobial potential. Several factors ...
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Introduction: Study in order to introduce new antimicrobial agents with natural origin to prevent the antibiotic resistance and eliminating its effects, employing chemical agents is an indispensable necessity. Honey, as an important food with natural origin has high antimicrobial potential. Several factors have contributed to the antibacterial activity of honey. For example, acidity, osmolarity, hydrogen peroxide, phenolic and flavonoid compounds, including these factors. Also, small amounts of glucose oxidase, protease, amylase, catalase and phosphatase enzymes and chemical compounds such as methylglyoxal are also effective. Floral origin of honey is effective on its biological properties including antimicrobial, anti-tumor, anti-inflammatory, antioxidant and antiviral activity. So this study was done to evaluate the antibacterial activity and physico-chemical analysis of four types of honey with different floral origin including: Thyme, Eryngium, Pennyroyal and Dill collected from the bee hives in Golestan province.
Materials and methods: Four honey samples with different floral origin including, Thyme, Eryngium, Pennyroyal and Dill were collected from the bee hive in the Golestan province in north of Iran. The bacterial strains used in this study, including two species of gram-negative of Escherichia coli and Shigella dysenteriae and the two species of gram-positive of Staphylococcus aureus and Bacillus cereus were provided in lyophilized. Bacterial strains in BHI broth were activated and from each of them were prepared bacterial suspensions equivalent to the McFarland 0.5 turbidity standard (1.5×108 CFU/ml). Evaluation of antibacterial activity using agar well diffusion method was performed. For this purpose serial dilutions of honey samples were prepared aseptically in sterilized distilled water. Surface of Mueller Hinton agar were uniformly inoculated with bacterial suspension containing of 1.5×108 CFU/ml. Then wells of 8 mm in diameter were prepared and these wells were filled with different dilutions of honey samples. After incubation at 37°C for 24 h, antibacterial activity was analyzed by measuring the zones of inhibition. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of honey samples were determined by using broth macro-dilution method. For this purpose, each of the tubes from different dilutions of honey samples were added by 5×105 CFU/ml from each of the tested bacteria and incubated for 24 h at 37°C. The results for microbial turbidity of visible were recorded. The last dilution (lowest concentration) in which microbial turbidity was not observed, as the MIC was considered. For the determination of MBC, from the tube that contained honey concentrations higher than the MIC were cultured onto the agar medium. The MBC was defined as the lowest concentration that allowed no visible growth on the agar. Also the studied physico-chemical properties were moisture content, pH, acidity, ash content and reducing sugars that was performed according to the Iranian National Standard No. 92.
Results and Discussion: The results indicated that the antibacterial effects of the tested honeys, and the difference between floral origins honeys is effective in antibacterial properties. MIC and MBC values obtained for the tested honeys were in the range of %6.25-50% (vol/vol). Highest antibacterial activity was recorded for Eryngium honey by agar well diffusion method with zone of inhibition of 15.5, 14,11 and 11 mm against S. dysenteriae, S. aureus, B. cereus and E. coli at concentration of 50% v/v respectively and its MIC for this bacteria were, 6.25%, 10%, 10% and 25% respectively. also low antibacterial activity of Pennyroyal honey was confirmed so that MIC of the this honey for E. coli, S. aureus, B. cereus and S.dysenteriae was, 50%, 25%, 12.5% and 10%, respectively. The quality of honey depends on a number of physico-chemical properties such as moisture, ash content, pH, acidity, the amount of sugars. For this reason, standards for honey have been set by different countries. The physico-chemical analysis of honeys showed moisture contents in the range of 22.56-25.36%, acidity in the range of 12.58-13.59 meq/kg, pH in the range of 4.15-4.26, reducing sugars in the range of 63.7-63.8%. Also the ash content of Eryngium honey with 0.183% was higher than the other honey samples. This higher level might be due to the higher pollen count in this region. High ash contents may also depend upon the floral origin of honey and the material collected by bees during foraging. The low acidity of honey samples studied in the present study was due to the fact that there was no unwanted fermentation in these samples. Overall, the results implied that honey samples with different floral origin collected from the bee hive in the Golestan province in north of Iran have variable potential antibacterial activity. The variety of antibacterial effects of different types of honey can be due to differences in plants that honey is obtained from. In other words, different species of plants in different regions have different compounds and their obtained honey will not be the similar and thus its biological effects also will be different.
