Antimicrobial activities of high molecular weight water‐soluble chitosans against selected gram‐negative and gram‐positive foodborne pathogens

Antimicrobial activities of high molecular weight water‐soluble chitosans (HMWWS) against selected Gram‐negative and Gram‐positive foodborne pathogens (initial inoculation of ca. 6.5 Log CFU mL^?1) were evaluated. Chitosans with 789 kDa and/or 1017 kDa were dissolved in aspartic acid (AS) to obtain 1–4% w/v solutions. Among HMWWS, only 4% 789 kDa AS chitosan reduced E. coli counts by 2 Log CFU mL^?1 from 7.33 at 0 h to 5.16 Log CFU mL^?1 at 96 h, and they were not effective against S. Typhimurium. Depending on the concentrations, HMWWS completely inhibited V. cholerae, V. vulnificus and V. parahaemolyticus as well as B. cereus and L. monocytogenes after 48 h or 96 h of incubation. Compared with the control (no HMWWS), 2% or 3% 1017 kDa AS chitosans showed about 3 Log CFU mL^?1 lower (4.72–4.86 vs. 7.71) for S. aureus at 96 h of incubation.     

Changes in growth kinetics and motility characteristics of Escherichia coli in the presence of sulphoraphane isolated from broccoli seed meal

In this investigation, the purified sulphoraphane from broccoli seed meal was evaluated in a range of 5–400 μg mL⁻¹ against Escherichia coli. The results showed that levels above 200 μg mL⁻¹ can inhibit 100% growth of 10⁶ CFU mL⁻¹ up to 5 h and cause a deformation of sigmoid behaviour in the growth kinetics, and even the bacterial population is reduced. On the other hand, low sulphoraphane levels such as 5–25 μg mL⁻¹ also have effects on the bacterium, such as the decrease in the maximum specific growth rate and the loss of ability to adjust to a growth model when maintained in isothermal conditions. In addition, sulphoraphane has an effect on swimming motility and swarming motility in semi-solid medium. The interaction of natural sulphoraphane–E. coli and the effect in the short periods can draw attention in research on food science and technology.     

Comparative study of the essential oils of three Beilschmiedia species and their biological activities

This study was designed to examine the chemical compositions of the essential oils from three Beilschmiedia species and antioxidant, antimicrobial, antityrosinase, acetylcholinesterase and anti‐inflammatory activities. The essential oils of B. kunstleri, B. maingayi, B. penangiana gave β‐caryophyllene (10.6–12.1%), β‐eudesmol (17.5–24.1%) and δ‐cadinene (17.5–28.7%) as the most abundant components respectively. The bark oil of B. maingayi showed the highest activity in β‐carotene/linoleic acid (125.9%) and phenolic content (288.2 mg GA g^?1), while B. penangiana bark oil was found to have strong activity in DPPH (IC₅₀ 84.7 μg mL^?1) and ABTS (IC₅₀ 108.3 μg mL^?1). The essential oils of B. penangiana showed the best activity against Candida glabrata with MIC value 31.3 μg mL^?1. The bark oil of B. penangiana gave 82.5% tyrosinase inhibiton. The leaf oil of B. maingayi gave the highest inhibition in AChE (66.6%) and lipoxygenase (77.0%) assay. Our findings demonstrate that the essential oils have great potential for applications in pharmaceutical and food industries.     

Mixed fermentation of jujube juice (Ziziphus jujuba Mill.) with L. rhamnosus GG and L. plantarum-1: effects on the quality and stability

This study was conducted aiming at improving the quality of jujube juice by mixed fermentation of lactobacilli. Selection of favourable lactobacilli and addition of nitrogen sources were explored as an efficient method to improve the viability of probiotic in jujube juice. After fermentation, the viability increased to 9.15 ± 0.10 Log CFU mL⁻¹, while the content of lactic acid increased to 5.61 ± 0.03 mg mL⁻¹. The total phenolic and total flavonoid content were 2663.03 ± 11.95 μg mL⁻¹ and 163.95 ± 0.47 μg mL⁻¹ respectively. Moreover, the stability of fermented jujube juice during refrigeration was investigated, which showed that the viability dropped to 8.84 ± 0.6 Log CFU mL⁻¹ and the concentration of lactic acid slowly increased to 6.51 ± 0.04 mg mL⁻¹; the ABTS value showed a 4.26% reduction and FRAP value did not significantly (P < 0.05) change during refrigerated storage. In addition to the existing knowledge, our data aid to the future applications of the jujube as a potential ingredient in novel probiotic foods formulation.     

Effect of carvacrol and thymol on Salmonella spp. biofilms on polypropylene

The present study evaluated the effects of carvacrol and thymol against Salmonella spp. biofilm on polypropylene. The efficacy of the compounds was assessed by quantifying Salmonella spp. cells during and after biofilm formation on polypropylene and performing scanning electron microscopy. During biofilm formation, carvacrol and thymol, at subinhibitory concentrations, reduced bacterial counts about 1–2 log, while established Salmonella spp. biofilms were reduced about 1–5 log by carvacrol and thymol, at MIC or 2× MIC. The greatest reduction in carvacrol‐treated biofilms, about 5 log, was observed with 156 and 312 μg mL^?1 (MIC and 2× MIC) in established Salmonella Typhimurium ATCC 14028 biofilms. Thymol showed the greatest reduction, about 4 log, at 624 μg mL^?1 (2× MIC) against mature Salmonella Enteritidis biofilm. Carvacrol and thymol reduced the number of Salmonella spp. cells on polypropylene, suggesting their potential for the control of Salmonella spp. biofilms.     

The prevention of fish spoilage by high antioxidant Australian culinary plants: Shewanella putrefaciens growth inhibition

Shewanella putrefaciens is a marine bacterium and a major microbial cause of spoilage in low temperature stored seafood. A survey of fruits and culinary herbs was undertaken on Australian plants with high antioxidant capacities. Twenty‐eight extracts from thirteen plant species were investigated for the ability to inhibit S. putrefaciens growth. Of these, eight extracts (28.6%) substantially inhibited S. putrefaciens growth. The muntries (Kunzea pomifera), lemon aspen (Acronychia acidula) and desert lime (Citrus glauca) extracts were efficient anti‐S. putrefaciens agents, with MIC values ≤3000 μg mL^?1. Of these, the muntries methanolic extract was the most potent growth inhibitor (MIC = 2240 μg mL^?1). The aqueous desert lime extract was also an effective growth inhibitor (MIC of 3857 μg mL^?1), whilst the methanolic bush tomato (Solanum aviculare), aqueous muntries and Davidson's plum (Davidsonia pruriens) extracts displayed moderate S. putrefaciens growth inhibition. All extracts were nontoxic in the Artemia fransiscana bioassay, with LC50 values (>1000 μg mL^?1). Nontargeted HPLC‐QTOF mass spectroscopy (with screening against three compound databases) putatively identified twenty compounds that were present in both inhibitory muntries extracts. The low toxicity of these extracts and their inhibitory bioactivity against S. putrefaciens indicates their potential as natural fish and seafood preservatives.     

Antibacterial mode of action of seed essential oil of Eleutherococcus senticosus against foodborne pathogens

This study investigated the antibacterial mechanism of action of the seed essential oil of Eleutherococcus senticosus (ESEO) against foodborne pathogenic bacteria. Preliminarily, the ESEO (1000 μg disc^?1) showed potential antibacterial effect as diameter of inhibition zones (12.0 ± 0.2–37.0 ± 2.0 mm) against the tested foodborne pathogens. The MIC and MBC values of ESEO against the tested bacteria were found in the range of 125–500 and 500–1000 μg mL^?1, respectively. At MIC concentration, the ESEO had potential inhibitory effect on the cell viability of the tested pathogens. In addition, SEM analysis showed the inhibitory effect of ESEO as confirmed by considerable morphological alterations on the cell wall of B. cereus ATCC 13061 and E. coli O157:H7 ATCC 43889. Moreover, the ESEO revealed its mode of action against foodborne pathogens on membrane integrity as confirmed by release of extracellular ATP, 260‐nm absorbing materials and leakage of potassium ions. These findings confirm that the ESEO can be used as a potential antibacterial agent in food industry to inhibit the growth of various foodborne pathogens.     

Effects of physicochemical factors and in vitro gastrointestinal digestion on antioxidant activity of R‐phycoerythrin from red algae Bangia fusco‐purpurea

R‐phycoerythrin (R‐PE) was purified from the red algae Bangia fusco‐purpurea after 35–50% ammonium sulphate fractionation followed by ion‐exchange column chromatography on DEAE‐Sepharose, resulting in a purity (A₅₆₅/A₂₈₀) ratio of 5.1. The circular dichroism spectroscopy results suggested that the structure of R‐PE is predominately helical. The antioxidant activity of R‐PE was studied and revealed changes in conformation and antioxidant activity at different temperatures and pH values. After in vitro‐simulated gastrointestinal (GI) digestion of R‐PE, the scavenging activity of ABTS radical (EC₅₀, 769.9 μg mL^?1), DPPH radical (EC₅₀, 421.9 μg mL^?1), hydroxyl radical (EC₅₀, 32.4 μg mL^?1) and reducing power (A₇₀₀ = 0.5, 625.8 μg mL^?1) were measured. Gel filtration chromatography analysis showed that the molecular weight distribution of the final GI digest that still contained high antioxidant activity was <3 kDa. Our present results indicate that digestion‐resistant antioxidant peptides of R‐PE may be obtained by in vitro GI proteinases degradation.     

Preparation and optimization of soy protein isolate–high methoxy pectin microcapsules loaded with Lactobacillus delbrueckii

This study was aimed to use soy protein isolate (SPI) and high methoxy pectin (HMP) as encapsulating materials for probiotic bacterial (Lactobacillus delbrueckii) delivery systems. The encapsulation conditions were optimised, and scanning electron microscopy (SEM) was used to characterise the microstructural changes of the microcapsule. The results showed that the optimal conditions for microcapsule preparation were 90 mg mL^?1 SPI and 1 mg mL^?1 HMP, with a SPI/HMP ratio of 7:1 (v/v), and a L. delbrueckii suspension to SPI–HMP complex ratio of 1:1 (v/v). The viability of the probiotics in the microcapsules reaching the small intestine was 3 log CFU mL^?1 higher than that of naked bacteria. SEM showed that the surface of the SPI–HMP compound microcapsules was smooth and that a large number of L. delbrueckii could be seen in cross‐sections of the microcapsules.     

Microwave‐assisted degradation of chitosan with hydrogen peroxide treatment using Box‐Behnken design for enhanced antibacterial activity

Low molecular mass (MM) chitosan with high degree of deacetylation (DDA) has excellent bioactivity including antioxidant, antibacterial and encapsulation properties. In this work, to reduce the MM of chitosan, microwave‐assisted heating treatment (MAHT) conditions were investigated using three factors at three levels Box‐Behnken design (BBD). Microwave heating (MH) time, H₂O₂ concentration and solid‐to‐liquid ratio significantly affected the DDA and MM of chitosan. The antibacterial activities of chitosan before and after degradation were investigated based on minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The results showed that a second‐order polynomial equation fitted the observed values using multiple regression analysis and had a high coefficient of determination (R² = 0.9591 and 0.9161 for the DDA and MM of chitosan, respectively). An optimisation study was performed using Derringer's desirability function methodology, and the optimal conditions were 80‐s MH time, 1.5% H₂O₂ concentration and 1:40 solid‐to‐liquid ratio. The MIC and MBC of chitosan before and after degradation against Escherichia coli and Salmonella typhimurium were 0.031 and 0.063 mg mL^?1, and 0.25 and 0.125 mg mL^?1, respectively. The optimised DDA and MM of chitosan were 90.58 ± 2.04% and 124.25 ± 14.36 kDa, respectively, which significantly reduces the use of oxidant reagent.     

Use of ultrasonication to enhance pea seed germination and microbial quality of pea sprouts

This study investigated the effects of ultrasonication, heating and chemical washing on decontamination of seeds of three pea (Pisum sativum) varieties and the microbial safety of their sprouts. Most of the decontamination treatments increased germination, decreased microbial load and enhanced sprout growth. Ultrasonication and NaOCl treatments reduced total aerobic counts on sprouts by 5.86 and 5.51 log units (averages of three varieties), respectively; while other chemical treatments (350 mL L^?1 ethanol, 350 mL L^?1 vinegar and 10 g L^?1 NaCl solutions) failed to reduce total aerobic counts sufficiently enough to secure microbial safety. The ultrasonication not only improved germination (increased from an average of 83 to 97%), but also reduced the total aerobic counts to 2.21 log₁₀ CFU g^?1 on seed sprouts. Ultrasonication also increased the sprouts yield to 301.83 g per 10 g seeds. Therefore, ultrasonication can be used as a microbial control treatment for pea sprout production.     

Phytochemicals content,antioxidant and hypoglycaemic activities of commercial nutmeg mace (Myristica fragrans L.) and pimento (Pimenta dioica (L.) Merr.)

Commercially dried powder of nutmeg mace (Myristica fragrans) and pimento (Pimenta dioica) spices was investigated for their high performance liquid chromatography phenolic profile and their antioxidant and hypoglycaemic properties by α‐amylase and α‐glucosidase inhibition tests. Generally, mace showed the most promising activity. An interesting protection of lipid peroxidation with an IC₅₀ value of 7.7 μg mL^?1 was found. A significant result was also obtained in ferric reducing ability power assay if compared to the positive control butylated hydroxytoluene (IC₅₀ value of 68.7 μg mL^?1 vs. 63.2 μg mL^?1, respectively). Mace also exhibited the highest carbohydrate‐hydrolysing enzyme inhibitory activity with IC₅₀ values of 62.1 and 75.7 μg mL^?1 against α‐amylase and α‐glucosidase, respectively. Overall, these results support the use of these spices not only as flavouring agent but also as food preservative and functional ingredients.     

Control of lactic acid bacteria in fermented beverages using lysozyme and nisin: test of traditional beverage boza as a model food system

The objective of this study was to increase quality and limited shelf‐life of boza (3–15 days), a traditional Balkan origin fermented beverage using lysozyme (LYS) and/or nisin (NIS). For this purpose, the effectiveness of NIS, LYS and LYS:NIS combinations was first tested in a broth medium at 4 °C for 3 weeks on Lactobacillus plantarum, one of the frequently isolated lactic acid bacteria in boza. Stability of LYS and NIS in boza, their effects on LAB counts, and chemical and sensory properties of boza were then evaluated during cold storage at 4 °C. Results of LAB counts as well as pH, d ‐ and l ‐lactic acid, and titratable acidity measurements showed that LAB in boza containing NIS (250 μg g^?1) or LYS:NIS (500:250 μg g^?1) could be controlled without reducing LAB counts below 6 log CFU mL^?1 during 2 weeks shelf‐life. In contrast, LYS (500 μg g^?1) alone could not control LAB in boza to delay its acidic spoilage. Positive effects of NIS and LYS:NIS application on quality of boza were also proved with sensory analysis by panelists and e‐nose measurements. This work showed that use of natural GRAS agents in preservation of fermented beverages containing probiotic LAB is possible without affecting their characteristic aroma and flavour.     

Synergistic effects of high pressure processing and slightly acidic electrolysed water on the inactivation of Bacillus cereus spores

Bacillus spores are concerns for their resistance to heat, high pressure processing (HPP), and disinfectants. We examined the effects of HPP and slightly acidic electrolysed water (SAEW) on inactivation of B. cereus spores. Spores' suspensions were prepared with 2‐(N‐morpholino) ethanesulfonic acid (MES) buffer or SAEW with available chlorine content (ACC) of 24, 35, 44 or 55 mg L^?1, and then subjected to HPP. The individual effects of HPP or SAEW on spores were negligible (<1.0 log CFU mL^?1). With factorial design and anova analysis, HPP + SAEW treatment was shown to have significantly positive effects on spores’ inactivation. The optimal conditions were 300 MPa HPP + SAEW with 44 mg L^?1 ACC or 200 MPa HPP + SAEW with 44 mg L^?1 ACC + 500 MPa HPP, producing reductions of 3.27 and 3.99 log CFU mL^?1, respectively. HPP + SAEW have potentials to serve as two effective hurdle techniques for inactivating B. cereus spores.     

Lactulose production from lactose by recombinant cellobiose 2‐epimerase in permeabilised Escherichia coli cells

Caldicellulosiruptor saccharolyticus cellobiose 2‐epimerase (CsCE) catalyses the single substrate lactose into lactulose and is the most efficient enzyme ever found for the enzymatic synthesis of lactulose. Ethanol‐permeabilised Escherichia coli cells containing CsCE were used as biocatalysts for lactulose production. The reaction conditions for maximum lactulose production were optimised to be 600 g L^?1 lactose, pH 7.5, 80 °C and 12.5 U mL^?1 of whole‐cell biocatalyst. After incubated at Na₂HPO₄–NaH₂PO₄ buffer for 2 h, approximately 390.59 g L^?1 lactulose was obtained with a conversion yield of 65.1%. The lowest production and conversion yield of epilactose were also found in Na₂HPO₄–NaH₂PO₄ buffer with a final concentration of 11.7 g L^?1 and a conversion yield <2%. The results represent a promising technology to attain high production and conversion yield of lactulose with a high purity on industrial scale.     

Membrane‐disruptive property of a novel antimicrobial peptide from anchovy (Engraulis japonicus) hydrolysate

A novel antimicrobial peptide named Pep39 has been isolated from anchovy hydrolysate using ÄKTA protein liquid chromatography and reverse‐phase high‐performance liquid chromatography. Its amino acid sequence was RLFRHAFKAVLRL with a molecular weight of 1626.92. Pep39 demonstrated antimicrobial ability against Escherichia coli ATCC 25922, Salmonella typhi ATCC 50013, Staphylococcus aureus ATCC 25923 and Bacillus subtilis ATCC 9372, with minimal inhibitory concentration (MIC) ranging from 4 to 32 μg mL^?1. No cytotoxicity to mouse red blood cells was observed when its concentration was lower than 30 μg mL^?1. Pep39 induced the influx of fluorescent probe 8‐Anilino‐1‐naphthalenesulfonic acid and the outflow of β‐galactosidase by increasing E. coli outer and inner membrane permeabilities, respectively. Flow cytometric analysis also demonstrated that Pep39 disrupted E. coli cell membrane. These results suggested that the antimicrobial mechanism of Pep39 involved cytoplasmic membrane damage.     

Effect of gums on viability and β‐galactosidase activity of Lactobacillus spp. in milk drink during refrigerated storage

The viability and β‐galactosidase activity of four Lactobacillus strains in milk drink containing gums during 28 days of refrigerated storage at 4 °C were assessed. The population of Lactobacillus rhamnosus GGB101 and Lactobacillus rhamnosus GGB103 were maintained, whereas the population of Lactobacillus reuteri DSM20016 and Lactobacillus reuteri SD2112 significantly decreased. The recommended level of 6 log CFU g^?1 was exceeded for all tested trains throughout storage. The highest viable number of Lactobacillus rhamnosus GGB103 (8.76 ± 0.03 log CFU mL^?1) was obtained in the product containing carrageenan–maltodextrin. The addition of guar–locust bean–carrageenan led to 20‐fold increase in the level of β‐galactosidase activity for L. rhamnosus GGB101 (1208 ± 2.12 Miller units mL^?1) compared to the control (61 ± 2.83 Miller units mL^?1). Our results suggested that gums could be added to milk to improve viability and enhance β‐galactosidase activity of Lactobacillus.     

Chemical composition,antioxidant activity and anti‐lipase activity of Origanum vulgare and Lippia turbinata essential oils

This study reported the chemical composition, phenolic content, antioxidant and anti‐lipase activity of oregano and Lippia essential oils. The major compounds found in oregano essential oil were γ‐terpinene (32.10%), α‐terpinene (15.10%), p‐cymene (8.00%) and thymol (8.00%). In Lippia essential oil, α‐limonene (76.80%) and 1,8‐cineole (4.95%) represented the major compounds. Oregano essential oil had higher phenolic content (12.47 mg gallic acid mL^?1) and DPPH scavenging activity (IC₅₀ 0.357 μg mL^?1) than Lippia essential oil (7.94 mg gallic acid mL^?1 and IC₅₀ 0.400 μg mL^?1, respectively). Both essential oils had similar antioxidant indexes (about 1.2) determined by Rancimat. Moreover, oregano essential oil had also higher anti‐lipase activity (IC₅₀ 5.09 and 7.26 μg mL^?1). Higher phenolic content in the essential oils was related with higher scavenging and anti‐lipase activities. Oregano and Lippia essential oils could be used as natural antioxidants on food products.     

Physical,antimicrobial and antibiofilm properties of Zataria multiflora Boiss essential oil nanoemulsion

It was evaluated physical, antibacterial and antibiofilm properties of Zataria multiflora Boiss essential oil (ZEO) and its nanoemulsion. Long‐term stability of nanoemulsion prepared by emulsion phase inversion was satisfying based on low narrow size distribution (polydispersity index ?0.2) and low droplet size (200 nm) over 21 days of storage. Nanoemulsion showed lower minimum inhibitory concentration (MIC) on Listeria monocytogenes (2500 µg mL^?1) than Salmonella Typhimurium (5000 µg mL^?1). Killing kinetics study revealed that nanoemulsion was more effective in inhibiting the growth of bacteria in milk than culture media. Both bacteriostatic and bactericidal effects were observed depending on the type of bacteria, nanoemulsion concentration and the time of exposure. Nanoemulsion at 4×MIC concentration reduced 64% and 75% of one‐day‐old biofilm of L. monocytogenes and S. Typhimurium, respectively. In conclusion, nanoemulsion revealed antimicrobial activity, but converting the ZEO to nanoemulsion did not improve its antibacterial activity; however, antibiofilm properties were enhanced.     

In vitro inhibitory effects of cranberry bean (Phaseolus vulgaris L.) extracts on aldose reductase, α‐glucosidase and α‐amylase

The in vitro inhibitory activities of different seed extracts prepared from cranberry bean mutant SA‐05 and its wild‐type variety Hwachia against aldose reductase, α‐glucosidase and α‐amylase were examined. The results indicated that the polyphenolics‐rich extracts obtained using 800 g kg^?1 methanol and 500 g kg^?1 ethanol demonstrated inhibitory activities against aldose reductase (IC₅₀ of 0.36–0.46 mg mL^?1) and α‐glucosidase (IC₅₀ of 1.32–1.94 mg mL^?1). The 500 g kg^?1 ethanol extracts also showed α‐amylase inhibitory activities (IC₅₀ of 70.11–80.22 μg mL^?1). Subsequent extracts, prepared further with NaCl and H₂O from precipitates of 800 g kg^?1 methanol or 500 g kg^?1 ethanol extracts, exhibited potent α‐amylase inhibitory activities (IC₅₀ of 17.68–38.68 μg mL^?1). A combination of 500 g kg^?1 ethanol extraction plus a subsequent H₂O extraction produced highest polyphenolics and α‐amylase inhibitors. The SA‐05 α‐amylase inhibitor extracts showed greater inhibitory activities than that of Hwachia. Thus, cranberry bean mutant SA‐05 is an advantageous choice for producing anti‐hyperglycaemic compounds.