Investigating the Effect of Melittin Peptide in Preventing Biofilm Formation, Adhesion and Expression of Virulence Genes in Listeria monocytogenes.

Listeria monocytogenes is a notable food-borne pathogen that has the ability to create biofilms on different food processing surfaces, making it more resilient to disinfectants and posing a greater risk to human health. This study assessed melittin peptide's anti-biofilm and anti-pathogenicity effects on L. monocytogenes ATCC 19115. Melittin showed minimum inhibitory concenteration (MIC) of 100 µg/mL against this strain and scanning electron microscopy images confirmed its antimicrobial efficacy. The OD measurement demonstrated that melittin exhibited a strong proficiency in inhibiting biofilms and disrupting pre-formed biofilms at concentrations ranging from 1/8MIC to 2MIC and this amount was 92.59 ± 1.01% to 7.17 ± 0.31% and 100% to 11.50 ± 0.53%, respectively. Peptide also reduced hydrophobicity and self-aggregation of L. monocytogenes by 35.25% and 14.38% at MIC. Melittin also significantly reduced adhesion to HT-29 and Caco-2 cells by 61.33% and 59%, and inhibited invasion of HT-29 and Caco-2 cells by 49.33% and 40.66% for L. monocytogenes at the MIC value. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) revealed melittin's impact on gene expression, notably decreasing inlB (44%) and agrA (45%) gene expression in L. monocytogenes. flaA and hly genes also exhibited reduced expression. Also, significant changes were observed in sigB and prfA gene expression. These results underscore melittin's potential in combating bacterial infections and biofilm-related challenges in the food industry.

The Effect of Plasma-Activated Water Combined with Rosemary Extract (Rosmarinus officinalis L.) on the Physicochemical Properties of Frankfurter Sausage during Storage.

This study investigated the impact of plasma-activated water (PAW) and rosemary extract on the bacterial inactivation and quality attributes of Frankfurter sausages during a 6-day storage period. The antibacterial activity, total phenol content (TPC), and total flavonoid content (TFC) of the rosemary extract were evaluated. The TPC of the rosemary extract was 89.45 mg gallic acid/g dry weight, while the TFC was 102.3 mg QE/g dry weight. Even at low concentrations, the rosemary extract effectively inhibited the growth of all the tested pathogens using the Well Diffusion Agar method (WDA). The sausages were treated with different concentrations of PAW and rosemary extract and stored for 1 and 6 days. Sample B (100% rosemary extract + PAW treatment) showed the greatest reduction in microbial load and was selected for further analysis. Throughout the storage period, Sample B exhibited no significant changes in pH, moisture content, textural parameters, or sensory evaluation compared to the control group. However, the hardness and color parameters (L*, a*) of Sample B decreased, while the TBARS value increased after 6 days of storage. The combination of PAW and rosemary extract, particularly Sample B, effectively inhibited bacterial growth in the Frankfurter sausages without compromising most quality attributes. Some changes in hardness, color, and lipid oxidation were observed over the extended storage period.

Identification of phytochemical, antioxidant, anticancer and antimicrobial potential of Calotropis procera leaf aqueous extract.

Since the dawn of civilization, people have turned to plants as a safe and efficient form of treatment for a variety of diseases. It has long been known that Calotropis procera has the potential to treat a number of diseases. In this study, the C. procera leaf aqueous extract was obtained using the maceration method, and p-coumaric was found to be the main compound. The extract was rich in phenols (174.82 mg gallic acid equivalent/g) and flavonoids (1781.7 µg quercetin equivalent/g). The extract had high antioxidant properties, as indicated by the IC50 values obtained for 2,2-diphenyl-1-picrylhydrazyl (DPPH) (366.33 µg/mL) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) (169.04 µg/mL), as well as the ferric ions reducing antioxidant power (FRAP) (1.67 µg ascorbic acid equivalent/g of the extract). The cytotoxicity of the extract was evaluated against the survival of HT 29 cells, and the IC50 was found to be 236.87 µg/mL. The most resistant and sensitive strains to the extract were Escherichia coli and Staphylococcus aureus, respectively. The morphological changes of these strains were demonstrated through scanning electron microscopy and confocal laser scanning microscopy. The C. procera extract could be therefore used as an antioxidant, antimicrobial, and anticancer agent.

Evaluation of the constituent compounds, antioxidant, anticancer, and antimicrobial potential of Prangos ferulacea plant extract and its effect on Listeria monocytogenes virulence gene expression.

Prangos ferulacea plant is very popular in Iran due to its unique properties in treating diseases and its special flavor. To check the characteristics of this plant, first, its extract was extracted using the maceration method. Its chemical composition was investigated using high-performance liquid chromatography (HPLC) that p-coumaric was identified as its main compound, and Fourier-transform infrared spectroscopy (FTIR) showed the presence of functional groups related to phenolic, flavonoid, tannins, and carboxylic acids such as caffeic acid and coumaric acid composition. Total phenol content (TPC), total flavonoid content (TFC), and beta-carotene were equal to 202.04 ± 5.46 mg gallic acid equivalent (GAE)/g dry weight, 1,909.46 ± 13 µg quercetin (QE)/g of dry weight, and 2.91 mg/100 g. The antioxidant property of the extract was evaluated using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) free radical scavenging and ferric reducing antioxidant power assay (FRAP). According to the IC50 obtained for DDPH (274 ± 7.2 µg/mL) and ABTS (120.45 ± 9.6 µg/mL) and FRAP values [1.92 ± 0.05 µg ascorbic acid equivalent (AAE)/g of extract], this extract had high antioxidant properties. Cytotoxicity was evaluated against the survival of HT 29 cells that IC50 was 82.15 ± 0.02 µg/mL. The antimicrobial property of the extract was calculated using disk diffusion agar (DDA), well diffusion agar (WDA), minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). Listeria monocytogenes has the highest sensitivity to this extract and inhibition zone based on DDA and WDA method and with an MIC and MBC equal to 16 and 128 mg/mL has the least resistance. The morphology change of L. monocytogenes strain was proved through scanning electron microscope (SEM) and confocal laser scanning microscopy (CLSM). The extract caused a significant reduction in the transcription of genes involved in the film formation ability of L. monocytogenes. The obtained results fully prove the very practical and pragmatic characteristics of P. ferulacea.

Evaluation of probiotic potential of autochthonous lactobacilli strains isolated from Zabuli yellow kashk, an Iranian dairy product.

AIMS: The aim of this study was to evaluate the probiotic potential and anti-biofilm activity of five lactobacilli strains which isolated and identified from an Iranian product. METHODS AND RESULTS: Five lactobacilli strains, which were isolated from Zabuli yellow kashk, were evaluated for the presence of probiotic properties, such as resistance to low pH, resistance to simulated gastrointestinal conditions, bile salt tolerance, hydrophobicity, and auto- and co-aggregation. In addition, antimicrobial susceptibility, adherence to Caco-2 cells (human colon cancer cell line), anti-adhesion activity, ability against biofilm formation and biofilm degradation of mentioned strains against Pseudomonas aeruginosa PTCC 1707 were assessed. All the strains tested showed acceptable characteristics, but Lactiplantibacillus plantarum TW57-4 appeared of particular interest. Some probiotic properties of this strain were similar and in some cases higher than the commercial probiotic strain Lacticaseibacillus rhamnosus GG (standard sample). Cholesterol assimilation and radical-scavenging activity of Lpb. plantarum TW57-4 were 70.2% and 62.3%, respectively. The adhesion degree of Lpb. plantarum TW57-4 was 10.6%. Applying competition and inhibition assay, this strain showed 55.3% and 62.3% of competition and inhibition activity in adhesion of P. aeruginosa PTCC 1707 to the intestinal cells, respectively. CONCLUSIONS: According to the obtained results, it can be concluded that Lpb. plantarum TW57-4 strain can be used as a promising candidate for in-vivo studies with the aim of developing new probiotic starter cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study furthers our understanding of lactobacilli strains behaviour after consumption to establish their beneficial effects.

Preparation and Functional Properties of Synbiotic Yogurt Fermented with Lactobacillus brevis PML1 Derived from a Fermented Cereal-Dairy Product.

Nowadays, production of functional foods has become very essential. Inulin is one of the most functional hydrocolloid compounds used in such products. In the present study, the production of a synbiotic yogurt containing 1, 2.5, and 5% (w/v) inulin has been investigated. The yogurt was fermented with Lactobacillus brevis PML1 derived from Tarkhineh, an Iranian cereal-dairy fermented food. Furthermore, the physicochemical properties, antioxidant activity, sensory attributes, and microbial viability properties were investigated on the 0th, 7th, and 14th days of storage after fermentation. The viable cells of L. brevis PML1 reached 108 CFU/g, and the product resisted to simulated digestive juices. Moreover, the synbiotic yogurt impressively increased the production of antimicrobial compounds and had the most profound antimicrobial effect on S. typhimurium. The physiochemical properties were in the normal range, and the fat content of the synbiotic yogurt was reduced remarkably. The antioxidant capacity of the fermented yogurt was significantly increased (p < 0.05), which was equal to those of DPPH (69.18 ± 1.00%) and BHA (89.16 ± 2.00%). The viability of L. brevis PML1 was increased during storage. Sensory analysis showed that there were significant differences in terms of the impressive parameters between the samples and the control (p < 0.05). Addition of 2.5% inulin not only improved the physical properties but also retained the viability of the probiotic after 14 days of storage, in addition to the viability of L. brevis with a viability count above 6 log CFU/g in the yogurt. Therefore, a novel synbiotic product containing L. brevis PML1, which can exert the desired properties, can be used as a suitable carrier for the delivery of the probiotic strain, exerting its beneficial health effects.

Optimization of gamma-aminobutyric acid production by Lactobacillus brevis PML1 in dairy sludge-based culture medium through response surface methodology.

Gamma-aminobutyric acid (GABA) is a pharmaceutical, bioactive amino acid that can produce by some species of Lactic Acid Bacteria (LAB). For the first time, we evaluated the production of GABA by Lactobacillus brevis PML1 in the medium that contain the contaminant food bio-product like dairy sludge and soybean meal. GABA production was analyzed by chromatography (TLC, HPLC) and the features of fermented extract which contains this amino acid were evaluated. The results of Response Surface Methodology (RSM) of Central Composite Design (CCD) at p < .05 showed 300 ppm of GABA production in optimal treatment including 14.77% dairy sludge powder, 6.27% soybean meal, and 0.49% ammonium sulfate (32°C for 120 hr fermentation). The results of fermented extract also showed the acceptable antimicrobial, antioxidant, and toxicity (against cancer cell) properties. Also, L. brevis PML1has not shown any hemolytic or DNase activity which confirm its safety aspects. According to the results, this new culture can be used as a cheap substrate to biological production of GABA, by L. brevis PML1 in various food and pharmaceutical formulations.

Control of microbial growth and lipid oxidation in beef using a Lepidium perfoliatum seed mucilage edible coating incorporated with chicory essential oil.

In this study, chicory essential oil (CEO) was obtained by hydrodistillation-based extraction method and it was rich in camphor (31.3%) and phenolic compounds with outstanding antioxidant and antimicrobial properties. The CEO was then incorporated into Lepidium perfoliatum seed mucilage (LPSM) based aqueous solution to prepare an active CEO-loaded LPSM edible coating. The effect of the edible coating was then investigated on the quality and shelf life of beef slices during 7 days storage at 4°C. The results revealed that beef slice coated with CEO-loaded LPSM edible coating had a significant inhibitory effect on its lipid oxidation and microbial growth. The CEO-LPSM coating also inhibited the weight and texture losses of beef slices during display more efficiently compared with the control and CEO-free LPSM coating. Besides, the beef slices coated with CEO-LPSM were the preferred samples in terms of sensory scores throughout the storage. Thus, using CEO-rich LPSM edible coating might inhibit decay and significantly improve the shelf life of fresh beef.

Gamma-aminobutyric acid production by Lactobacillus brevis A3: Optimization of production, antioxidant potential, cell toxicity, and antimicrobial activity.

In this study, whey powder was used as the basic compound for fermentation culture and the production of bioactive gamma-aminobutyric acid (GABA) compound. GABA is a nonprotein four-carbon amino acid that inhibits stress signals by preventing brain signals, reducing stress, and being effective in treating neurological disorders and decreasing the growth of cancer cells. Due to the side effects caused by the chemical type of GABA, the biological production of GABA has attracted. Three levels of whey powder (5%, 10%, and 15%), and monosodium glutamate (MSG) (1%, 3%, and 5%) were selected at temperatures (25, 30, and 37°C) and after fermentation, the presence of GABA in the culture medium was examined by thin-layer chromatography. The optimal amount of GABA was measured by using high-performance liquid chromatography. The results of the central composite design of the response surface methodology at a significant level of 95% showed that the optimal treatment was 14.96% whey powder, 4.95% MSG at temperature of 37°C and fermentation for 48 hr and under these conditions, GABA production was 553.5 ppm. The results of the fermented extract tests showed that the highest antimicrobial activity was on Escherichia coli and the highest free radical scavenging was 59.67%. The IC50 level in the Caco-2 cancer cell cytotoxicity test was 39.5 mg/ml. According to the results, the combination of whey with MSG can be used as a cheap substrate to produce a valuable bioactive GABA product, and the cellular extract of this fermentation can also be used as an antimicrobial and antioxidant compound in food and pharmaceutical formulations.

Oral Immunotherapy Using Probiotic Ice Cream Containing Recombinant Food-Grade Lactococcus lactis Which Inhibited Allergic Responses in a BALB/c Mouse Model.

This study was conducted to evaluate the effects of recombinant probiotic bacteria as a candidate for oral vaccine with the potential of treating allergy to Amaranthus retroflexus pollens. The main gene of this allergen, Ama r 2, was cloned into the food grade plasmid pNZ7025 and then was electrotransformed into the food grade Lactococcus lactis NZ1330. No expression was observed in the primary structure due to the distance between the ribosome binding site and the start codon. Therefore, the vector structure was corrected using the site-directed mutagenesis (SDM) technique. The cell extract of this strain was used for assessing the expression of the recombinant allergen in western blot analysis, and the existence of this protein with a molecular weight of 14.2 kDa was confirmed. To evaluate the efficacy of this strain in the treatment of allergies as an oral vaccine, probiotic ice cream was prepared. After the sensitization of mice, the treatment was performed by oral immunotherapy for 4 weeks, 4 to 5 times per week. 20 µl of functional ice cream with 1012 CFU/ml of r-L. lactis NZ1330 significantly reduced the serum IgE level. The levels of IFN-γ and TGF-ß cytokines increased in the 20 µl ice cream treatment group as well as 40 µg/ml pure allergen compared with the PBS-treated group, and IL-4 cytokine levels decreased compared with the PBS-treated group. Overall, 20 µl ice cream with 1012 CFU/ml of the recombinant bacteria resulted in the best performance in terms of improving allergies to Th1 and Treg responses.

Probiotic characterization of Pediococcus strains isolated from Iranian cereal-dairy fermented product: Interaction with pathogenic bacteria and the enteric cell line Caco-2.

In present study, we investigated the probiotic potential of three Pediococcus spp. isolated from Iranian traditional fermented cereal-dairy product, Tarkhineh. These 3 strains were identified as Pediococcus acidilactici VKU2, P. acidilactici IAH-5 and P. pentosaceus DHR005 by 16S rRNA gene sequencing. All the strain were found tolerate to pH 3 and 0.3% oxall for 3 h as well as simulated gastric and intestinal juice. P. acidilactici IAH-5 showed the highest cholesterol removal (67.52%), hydroxyl radical scavenging activity (58.32%), hydrophobicity (40.3%) and auto-aggregation (48%). Pediococcus spp. inhibited the growth of tested pathogens (Escherichia coli ATCC 25922, Pseudomonas aeruginosa PTCC 1707, Salmonella typhimurium PTCC 1609, and Staphylococcus aureus ATCC 25923) which the most susceptible strain was S. aureus. In competition assay, P. acidilactici IAH-5 was able to inhibited adhesion of 67.3% of S. typhimurium and in inhibition assay 45.8% of the pathogenic adhesion to Caco-2 cells were decreased. P. acidilactici VKU2 and P. acidilactici IAH-5 showed 16.32 and 12.25% adhesion to simulated epithelial cell line which were also confirmed by scanning electron microscopy. Pediococcus spp. did not showed DNase production or hemolytic activity which confirm its safety aspects. Our findings suggested that the P. acidilactici IAH-5 has the best properties with probiotic features and cholesterol assimilation for its application as novel bio-therapeutic and bio-preservation agents.

Analysis of different signal peptides for the secretory production of Ama r 2 in gram-positive systems (Lactococcus lactis).

Prokaryotic systems have been considered the most affordable and simplest hosts which are being employed to express recombinant proteins such as allergens; nevertheless, without appropriate signal peptide (SP), these systems cannot be used for secretory proteins. Recently, a lot of effort has been put into assessing the potential of gram-positive strains such as lactic acid bacteria for new applications in the production of heterologous proteins. Ama r 2 is a respiratory allergen from Amaranthus retroflexus, whose recombinant production in the probiotic host could be introduced as a specific and effective way to rapid diagnosis and immunotherapy of this allergy. Consequently, the production of this recombinant protein using the prokaryotic system, requires a suitable SP to protect disulfide bonds and to prevent misfolding. This study was designed to predict the best SPs for the expression of Ama r 2 protein in Lactococcus lactis as the host. In this study, 42 signal sequences were selected from SP databases and the most important features of them were evaluated. First, n, h and c regions of the SPs and their probabilities were investigated by signalP software version 4.1. Then, their physicochemical properties were evaluated by Portparam and SOLpro. Moreover, the secretion sorting and sub-cellular localization sites were evaluated by PRED-TAT and ProtcompB software programs. The results revealed that yjgB, entC2 (Entrotoxine type C-2), ent B (Entrotoxine type), blaZ (Beta lactamase), dex (number 21), blm (Beta lactamase 2), dex (Dextranase; number 20) and number 26 were introduced theatrically as the best SPs to express Ama r 2 in Lactococcus lactis.

Antagonistic activity of recombinant Lactococcus lactis NZ1330 on the adhesion properties of Escherichia coli causing urinary tract infection.

Death from infectious diseases has caused concerns about increases in the resistance of pathogens, impelling researchers to create novel therapeutic solutions. The management of intestinal tract problems has been the advance use of probiotics in medicine. The aim of this study was evaluate the physicochemical cell surface and adhesion properties of recombinant Lacotococcus lactis NZ1330 containing Ama r 2 gene, followed by the assessment of the antagonistic activity of this strain against the Escherichia coli causing urinary tract infection (UTI) in humans. For this purpose, cloning and expression of Ama r 2 gene were done. Afterwards, acid and bile resistance, which are the primary characteristics of any probiotic, were evaluated. The r-L. lactis NZ1330 was examined for the physicochemical properties of cell surfaces and the adhesion properties against Escherichia coli. Furthermore, the potential of the recombinant strain to adhere to adenocarcinoma intestinal cell line, Caco-2 cells, as well as the antagonistic properties of r-L. lactis NZ1330 against E. coli was investigated. r-L. lactis NZ1330 was capable of surviving at low pH and different concentrations of bile salts. 40.1% hydrophobicity, 36.5% auto-aggregation and 14.4% co-aggregation were observed for this strain. The adhesion level of r-L. lactis NZ1330 was 5.7% which was also confirmed by scanning electron microscopy (SEM). r-L. lactis NZ1330 was able to compete, inhibit and displace the adhesion of Escherichia coli to Caco-2 cells. r-L. lactis NZ1330 was considered to be a reliable probiotic alternative by showing these desirable properties. Results revealed that Ama r 2 gene expression had no effect on the positive probiotic properties of L. lactis NZ1330, proving this strain could be a suitable probiotic host for the expression of this allergen.

Evaluation of adherence and anti-infective properties of probiotic Lactobacillus fermentum strain 4-17 against Escherichia coli causing urinary tract infection in humans.

Gastrointestinal (GI) infection is one of the most common types of infectious diseases. Application of probiotic strains in the control of such infections represents a promising approach. In this study, Lactobacillus fermentum strain 4-17 was isolated from kashkineh, an Iranian cereal fermented food, and identified by sequencing its 16S rRNA gene using universal primers. Its probiotic features, including resistance to acid, bile tolerance, antibacterial activity, resistance to intestinal and gastric juices, and hydrophobicity were evaluated. The ability of this strain to adhere to human intestinal cells as well as the anti-adhesive effect of L. fermentum strain 4-17 against E. coli isolated from patients with urinary tract infection was investigated. L. fermentum strain 4-17 was capable of surviving at various conditions such as low pH values, bile salts exposure, and GI tract environment. It showed 43% cell hydrophobicity. The adhesion level of L. fermentum strain 4-17 to human colon adenocarcinoma Caco-2 cells was 8.5% which was also confirmed by scanning electron microscopy (SEM). Furthermore, this strain showed appropriate anti-adhesive properties (including competition, inhibition and replacement properties) against human pathogenic bacteria. These data suggest that L. fermentum strain 4-17 could be examined further for its useful effects and introduced as a novel candidate probiotic to control GI infection disease.

Heterologous expression of Thrombocidin-1 in Pichia pastoris: Evaluation of its antibacterial and antioxidant activity.

The antimicrobial peptide Thrombocidin-1 (TC-1) isolated from human blood that derived from NAP-2 by deleting of two amino acids from C-terminal region. In this study, a C-terminal 6 _ His tagged recombinant TC-1 was expressed as a secreted peptide in Pichia pastoris, for the first time. The recombinant P. pastoris was inoculated in to BMMY culture medium, incubation with 5 µl/ml absolute methanol for 72 h at 30 °C. The TC-1 peptide was concentrated with nickel affinity chromatography and electrophoresis on 16% acrylamide gels. The molecular weight of recombinant TC-1 is approximately 8 kDa and under these conditions, the concentration of TC-1 considered 190 µg/ml that determined by the Bradford method. The antimicrobial activity test (Minimum Inhibitory Concentration and Minimum Bactericidal Concentration) was done against: Listeria monocytogenes, Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa. The growth of these pathogenic bacteria was limited when we used peptide at a concentration of as low as 19.56 µg/ml. Based on DPPH radical scavenging (DPPH-RS) activity and reducing power assays, this peptide showed relatively good antioxidant potential in comparison with standard antioxidant used in this study (BHT). Due to the existence of TC-1 in blood, which makes it safe for human consumption, and the good results of its antimicrobial and antioxidant activity, it can be introduced as a good alternative and a novel effective peptide to food industry for bio-preservation.

Phytochemical analysis and antibacterial activities extracts of mangrove leaf against the growth of some pathogenic bacteria.

In this study, the effects of water, ethanol, methanol and glycerin at five levels (0, 31.25, 83.33, 125 and 250 ml) were investigated on the efficiency of mangrove leaf extraction using mixture optimal design. The antimicrobial effect of the extracts on Streptococcus pneumoniae, Enterococcus faecium and Klebsiella pneumoniae was evaluated using disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) methods. The mangrove leaf extraction components were identified through gas chromatography/mass spectrometry (GC/MS). Phytochemical analysis (alkaloids, tannins, saponins, flavone and glycosides) were evaluated based on qualitative methods. Antioxidant activity of extracts was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant potential (FRAP) methods. Maximum antimicrobial effect was observed in Enterococcus faecium and highest resistance against mangrove leaf extract in Enterococcus faecium and Klebsiella pneumoniae, respectively. Increasing concentration of mangrove extracts had a significant effect (p ≤ 0.05) on inhibition zone diameter. The MICs of the mangrove leaf extraction varied from 4 mg/ml to 16 mg/ml. The optimum formulation was found to contain glycerin (0 ml), water (28.22 ml), methanol (59.83 ml) and ethanol (161.95 ml). The results showed that the highest antioxidant activity was related to optimum extract of mangrove leaf and ethanolic extract respectively. The results of phytochemical screening of Avicennia marina leaves extract showed the existence of alkaloids, tannins, saponins, flavone and glycosides. 2-Propenoic acid, 3-phenyl- was the major compound of Avicennia marina. The results of non-significant lack of fit tests, and F value (14.62) indicated that the model was sufficiently accurate. In addition, the coefficient of variations (16.8%) showed an acceptable reproducibility.

Oliveria decumbens essential oil: Chemical compositions and antimicrobial activity against the growth of some clinical and standard strains causing infection.

Oliveria decumbens as a valuable medicinal plant is extensively used in traditional medicine. clinical and standard strains causing infection resistance to antimicrobial agents, is one of the important problems in medicine. The aim of this study was to investigate the antibacterial activities and phytochemical analysis of Oliveria decumbens essential oil on the growth of some clinical and standard strains causing infection (Pseudomonas aerogenes, Escherichia coli, Streptococcus pyogenes and Staphylococcus epidermidis). Oliveria decumbens essential oil composition was identified by gas chromatography/mass spectrometry. Phytochemical analysis (alkaloids, saponins, flavone and phenolic) essential oil of the Oliveria decumbens were appraised based on qualitative methods. Several methods (disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)) were used to appraise the antibacterial activity of the Oliveria decumbens essential oil. Thymol (28.45%) was the major compound of Oliveria decumbens essential oil. The total phenolics content (TPC) of the essential oil positively correlated with antioxidant activity (AA). The TPC and AA of Oliveria decumbens essential oil was equal to 92.45 ± 0.70 µg GAE/mg and 164.45 ± 1.20 µg/ml, respectively. The MIC of Oliveria decumbens essential oil ranged from 1 to 8 mg/ml depending on the type of bacteria (clinical and standard strains). The MBC of Oliveria decumbens essential oil varied from 1 mg/ml to 16 mg/ml. The smallest inhibition zone diameter (IZD) on different Oliveria decumbens essential oil concentrations on P. aeruginosa. Results indicate that Oliveria decumbens essential oil can prove to be an important source of AA and antibacterial and may be used for the treatment of infection diseases.

Diversity and Probiotic Potential of Lactic Acid Bacteria Isolated from Horreh, a Traditional Iranian Fermented Food.

The aim of this study was to evaluate the probiotic potential of lactic acid bacteria (LAB) strains isolated from Horreh. Some probiotic properties, e.g., resistance to acid, bile tolerance, antibacterial activity, and antibiotic susceptibility, were investigated. A total of 140 Gram-positive and catalase-negative isolates from Horreh were subjected to identification and grouping by cultural methods and the 16S rRNA sequencing. The new isolates were identified to be Lactobacillus (fermentum, plantarum, and brevis) Weissella cibaria, Enterococcus (faecium and faecalis), Leuconostoc (citreum and mesenteroides subsp. mesenteroides) and Pediococcus pentosaceus. Probiotic potential study of LAB isolates showed that Lb. plantarum and Leu. mesenteroides subsp. mesenteroides isolates were able to grow at pH 2.5 and 3.5. Lactobacillus plantarum (isolate A44) showed the highest cell hydrophobicity (84.5%). According to antibacterial activity tests, Listeria innocua and Staphylococcus aureus were the most sensitive indicators against the selected LAB strains, while Escherichia coli and Bacillus cereus were the most resistant. In addition, all the isolated LAB species were resistant to vancomycin. The results of the present study suggested that the Lactobacillus fermentum and plantarum isolated from Horreh, characterized in this study, have potential use for industrial purposes as probiotics.

Optimization of the production conditions of the lipase produced by Bacillus cereus from rice flour through Plackett-Burman Design (PBD) and response surface methodology (RSM).

In this study, the screening of lipase positive bacteria from rice flour was carried out by Rhodamin B agar plate method. Bacillus cereus was identified by 16S rDNA method. Screening of the appropriate variables and optimization of the lipase production was performed using Plackett-Burman design (PBD) and response surface methodology (RSM). Among the isolated bacteria, an aerobic Bacillus cereus strain was recognized as the best lipase-producing bacteria (177.3 ± 20 U/ml). Given the results, the optimal enzyme production conditions were achieved with coriander seed extract (CSE)/yeast extract ratio of 16.9 w/w, olive oil (OO) and MgCl2 concentration of 2.37 g/L and 24.23 mM, respectively. In these conditions, the lipase activity (LA) was predicted 343 U/mL that was approximately close to the predicted value (324 U/mL), which was increased 1.83 fold LA compared with the non-optimized lipase. The kinetic parameters of Vmax and Km for the lipase were measured 0.367 µM/min.mL and 5.3 mM, respectively. The lipase producing Bacillus cereus was isolated and RSM was used for the optimization of enzyme production. The CSE/yeast extract ratio of 16.9 w/w, OO concentration of 2.37 g/L and MgCl2 concentration of 24.23 mM, were found to be the optimal conditions of the enzyme production process. LA at optimal enzyme production conditions was observed 1.83 times more than the non-optimal conditions. Ultimately, it can be concluded that the isolated B. cereus from rice flour is a proper source of lipase.