Combination Of Nisin Research Articles

In vitro synergistic activities of cefazolin and nisin A against mastitis pathogens.

First-generation cephalosporins such as cefazolin (CEZ) have been widely used for mastitis treatment in dairy cattle. However, the use of antibiotics results in the presence of antibiotic residues in milk, which is used for human consumption. Nisin A, a bacteriocin produced by Lactococcus lactis, has been used as a broad-spectrum food preservative for over 50 years. Therefore, a combination of CEZ and nisin A might provide an extended activity spectrum against mastitis pathogens and reduce the antibiotic dose for mastitis treatment. This study aimed to evaluate the combined effect of CEZ and nisin A against mastitis pathogens using the checkerboard and time-kill assays. In the checkerboard assay, the CEZ-nisin A combination exhibited a synergistic effect against Staphylococcus aureus (n=20/20) and Enterococcus faecalis (n=13/18), and meanwhile exhibited a mostly additive effect against Staphylococcus intermedius (n=12/20), Streptococcus agalactiae (n=10/10), Streptococcus dysgalactiae (n=18/18), and Escherichia coli (n=14/18). There were no indifferent or antagonistic effects between CEZ and nisin A. In the time-kill assay, the CEZ-nisin A combination at 0.5 × or 1 × minimum inhibitory concentration exhibited synergistic reduction of bacterial growth by over 3 log10 colony forming units per ml relative to that observed with either antimicrobial substance alone. These results suggest that the CEZ-nisin A combination can be used for developing an intramammary infusion for mastitis treatment, with lower antibiotic concentrations than normal.

Enterocin B3A-B3B produced by LAB collected from infant faeces: potential utilization in the food industry for Listeria monocytogenes biofilm management.

Enterococcus faecalis B3A-B3B produces the bacteriocin B3A-B3B with activity against Listeria monocytogenes, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA) and Clostridium perfringens, but apparently not against fungi or Gram-negative bacteria, except for Salmonella Newport. B3A-B3B enterocin has two different nucleotides but similar amino acid composition to the class IIb MR10A-MR10B enterocin. B3A-B3B consists of two peptides of predicted molecular mass of 5176.31Da (B3A) and 5182.21Da (B3B). Importantly, B3A-B3B impeded biofilm formation of the foodborne pathogen L. monocytogenes 162 grown on stainless steel. The antimicrobial treatment of stainless steel with nisin (1 or 16mgml-1) decreased the cell numbers by about 2 log CFUml-1, thereby impeding the biofilm formation by L. monocytogenes 162 or its nisin-resistant derivative strain L. monocytogenes 162R. Furthermore, the combination of nisin and B3A-B3B enterocin reduced the MIC required to inhibit this pathogen grown in planktonic or biofilm cultures.

Combination of nisin and ε-polylysine with chitosan coating inhibits the white blush of fresh-cut carrots

Effects of the combination of nisin and ε-polylysine with chitosan coating on quality maintenance and white blush inhibition were investigated in fresh-cut carrots. Fresh-cut carrots were treated with 1% lactic acid solution (v/v), 1% chitosan solution (w/v), or 1% chitosan solution containing 64 μg/mL nisin and 250 μg/mL ε-polylysine (LA + CH + Nisin + ε-PL). The samples were packed in polyethylene plastic bags and stored at 4 °C for 9 days. Changes in sensory attributes, physicochemical indices, respiration rate, microbiological counts and white blush were measured. Results showed that LA + CH + Nisin + ε-PL significantly (P < 0.05) inhibited respiration rate, decline of ascorbic acid and growth of microorganism (yeast and mold, total viable counts, total coliforms counts, Staphylococcus aureus and Pseudomonas spp.), and increased total phenol content and phenylalanine ammonialyse (PAL) activity compared with the control after 9-day storage. It was also strongly effective in inhibiting the white blush of fresh-cut carrots. Furthermore, LA + CH + Nisin + ε-PL significantly (P < 0.05) reduced the lignin synthesis in fresh-cut carrots by inhibiting the cinnamate-4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL) activity, as well as Dc4CL and DcC4H gene expression. Our results may provide some basis for the use of the combination of nisin and ε-polylysine with chitosan coating as an alternative preservation method for fresh-cut carrots.

The Effect of RF Treatment Combined with Nisin Against Alicyclobacillus Spores in Kiwi Fruit Juice

In the present study, the efficiency of the treatment of radio frequency (RF) combined with nisin against the spores of three strains of Alicyclobacillus from two species was examined with the response surface methods (RSM). The results showed that the combination of RF and nisin was effective on killing Alicyclobacillus acidoterrestris and Alicyclobacillus contaminans spores in kiwi fruit juice. The killing effect increased with the increase of temperature, holding time, and nisin concentration, but the increment was not significant at high levels of temperature and nisin concentration. The killing effect between A. acidoterrestris and A. contaminans strains was different in kiwi fruit juice. The strain with the highest killing effect was DSM 17975T, followed by DSM 3923T and C-ZJB-12-33. The reduction of spore number of DSM 17975T at the central design points of RSM was about 2.4. Under experimental conditions, nisin showed hardly any effect on killing DSM 17975T spores when combined with RF.

Biocontrol of Listeria monocytogenes ATCC 7644 on fresh-cut tomato (Lycopersicon esculentum) using nisin combined with organic acids.

The biocontrol of Listeria monocytogenes on fresh-cut tomato using nisin and organic acids was investigated. Fresh-cut samples inoculated with 108 CFU/mL of L. monocytogenes, treated with nisin (5,000 IU/mL), a combination of nisin and organic acids (acetic and citric acids at 3 and 5% each), with chlorine at 200 ppm as a control, and stored for six days at 4, 10, and 25°C were used to evaluate certain physicochemical qualities (pH, titratable acidity, soluble solid content, vitamin C content, and color). Nisin treatment significantly (p<0.05) reduced bacterial population by 1.91-3.07 log CFU/mL. Nisin-citric acid combination provided 2.65-3.29 log CFU/mL reduction, while nisin-acetic acid combination provided 2.93-4.15 log CFU/mL reduction. The control treatment provided <1-2 fold log reductions. Slight variations in physicochemical properties of fresh-cut tomato were observed. Nisin and organic acids can be used to improve the microbial safety of fresh-cut tomato.

Effects of Colistin and Bacteriocins Combinations on the In Vitro Growth of Escherichia coli Strains from Swine Origin.

Escherichia coli strains from swine origin, either susceptible or resistant to colistin, were grown under planktonic and biofilm cultures. After which, they were treated with antibacterial agents including nisin and enterocin DD14 bacteriocins, colistin and their combinations. Importantly, the combination of colistin, enterocin DD14 and nisin eradicated the planktonic and biofilm cultures of E. coli CIP54127 and the E. coli strains with colistin-resistance phenotype such as E. coli 184 (mcr-1 +) and E. coli 289 (mcr-1 -), suggesting therefore that bacteriocins from lactic acid bacteria could be used as agents with antibiotic augmentation capability.

Comparative study of the microencapsulation by complex coacervation of nisin in combination with an avocado antioxidant extract

The objective of this study was to evaluate several conditions for complex coacervation to encapsulate a mixture of nisin, an antimicrobial peptide, and an antioxidant avocado peel extract. We evaluated the effects of three factors: two matrix-wall systems (collagen-alginate and collagen-pectin), two drying methods (freeze and spray drying) and two core dispersion systems (water in oil emulsion or in suspension). The final characteristics measured for each microencapsulation treatment were: encapsulation efficiency, encapsulation loading, encapsulation yield, Aw, and moisture. The core dispersion method (emulsified or not) and drying method (freeze or spray), as well as their interactions, were important factors in the final characteristics of the microencapsulation produced by complex coacervation. The interaction of the three factors only had an effect on moisture. The core dispersion factor had an effect on the encapsulation loading of both bioactives, whereas the drying method had an effect on Aw and encapsulation yield. Higher loading was obtained with non-emulsified core; higher encapsulation efficiency and yield were obtained through the combination of spray drying with emulsified core. Higher Aw, and moisture were obtained with the freeze drying method and emulsified core. The drying methods modified the final morphology, shape and structure of the microcapsules.The wide range of microcapsules produced by the combination of the three assessed factors may offer the food industry a wide range of option to deliver functional ingredients and meet various types of needs.

Synergistic interactions of nisin in combination with cinnamaldehyde against Staphylococcus aureus in pasteurized milk

Staphylococcus aureus (S. aureus) is the leading cause of a broad range of human infections and associated with food poisoning worldwide. Nisin can be considered as a natural preservative and is widely used by the food industry. However, resistance of nisin has been reported in several species of bacteria. Identification of antibacterial agents that are synergistic with nisin is a method of minimising resistance to nisin. Cinnamaldehyde (CA), a well-studied essential oil isolated from cinnamon bark, has been reported to exhibit a broad spectrum of antimicrobial activity across a wide range of microorganisms. Previous studies have demonstrated that CA could be used in the food industry due to its active antibacterial activity and its safety. In this study, the synergistic antibacterial activity in vitro of nisin and CA against 13 food-borne isolates of S. aureus and S. aureus ATCC 29213 were evaluated by a checkerboard assay. The FICI values of nisin combined with CA ranged from 0.1875 to 0.375, which demonstrated a synergistic effect. The positive interactions were verified using the challenge tests in pasteurized milk and agar diffusion assays. The LIVE/DEAD BacLight experiment and scanning electron microscope (SEM) assay were also performed to distinguish the intact and damaged both of cell walls and membranes with the compounds, the combination group displayed stronger damage both of cell wall and cell membrane than when nisin or CA was used alone. These findings indicated that the combination of nisin and CA could not only be used as a promising naturally sourced food preservative but also to reduce the problem of bacterial resistance.

Levels of phenolic compounds, antioxidant capacity, and microbial counts of fresh-cut onions after treatment with a combination of nisin and citric acid

Nisin and citric acid are generally recognized as safe (GRAS) for using as a food ingredient and often used to control the microbial contamination of fresh-cut fruits and vegetables. However, their effects on phenolics and antioxidants remain unclear. Here we used 50 μg·mL-1 nisin combination with 1% (w/v) citric acid (N+C) to wash fresh-cut onions and the phenolic compounds, antioxidant capacity, microbial load, slice color were evaluated during the storage period. N+C treatment decreased the total polyphenols (TP) and quercetin contents (p < 0.05), as compared to the control at day 0, but did not affect phenolic acids and antioxidant capacity (ABTS, DPPH, and FRAP assays). During storage, antioxidant capacity, TP, and flavonoid contents increased. Storage had mixed effects on the phenolic acids (caffeic acid, vanillic acid, and ferulic acid increased, chlorogenic acid remained fairly stable). N+C treatment reduced the microbial counts and total viable counts (TVC) of fresh-cut onions below the detection limit until 5 days of storage. After 15 days storage, TVC of N+C treated samples remained significantly (p < 0.05) lower than controls. In addition, N+C treatment led to better color retention during storage. Therefore, N+C might be used as a safe preservative for fresh-cut onions.

Antimicrobial Activities of Nisin, Tea Polyphenols, and Chitosan and their Combinations in Chilled Mutton.

Antimicrobial activities of nisin, tea polyphenols (TPs), and chitosan, and their combinations were evaluated against both Gram-positive bacteria (GPB) and Gram-negative bacteria (GNB) by the agar dilution method. Results showed that the MIC of nisin was 2.44 to 1250 mg/L for GPB and reached 5000 mg/L for GNB. The MICs of TPs and chitosan were 313 to 625 mg/L and 469 mg/L for GNB, and 156 to 5000 mg/L and 234 to 938 mg/L for GPB, respectively. These results indicated that TPs and chitosan exhibited inhibitory effects against both GPB and GNB, whereas nisin inhibited the growth of GPB only. Based on the orthogonal test of their MICs, and evaluation of preservative effect and sensory attributes in chilled mutton, the optimum combination was chosen as 0.625, 0.313, and 3.752 g/L for nisin, TPs, and chitosan, respectively. By using the optimum treatment, the shelf life of chilled mutton was extended from 6 to 18 d at 4 °C in the preservative film packages. These results indicate that the combination of nisin, TPs, and chitosan could be used as preservatives to efficiently inhibit the growth of spoilage microorganisms and pathogens in meat, thus improving the safety and shelf life of chilled mutton.

In Vitro Activities of Nisin and Nisin Derivatives Alone and In Combination with Antibiotics against Staphylococcus Biofilms.

The development and spread of pathogenic bacteria that are resistant to the existing catalog of antibiotics is a major public health threat. Biofilms are complex, sessile communities of bacteria embedded in an organic polymer matrix which serve to further enhance antimicrobial resistance. Consequently, novel compounds and innovative methods are urgently required to arrest the proliferation of drug-resistant infections in both nosocomial and community environments. Accordingly, it has been suggested that antimicrobial peptides could be used as novel natural inhibitors that can be used in formulations with synergistically acting antibiotics. Nisin is a member of the lantibiotic family of antimicrobial peptides that exhibit potent antibacterial activity against many Gram-positive bacteria. Recently we have used bioengineering strategies to enhance the activity of nisin against several high profile targets, including multi-drug resistant clinical pathogens such as methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, staphylococci, and streptococci associated with bovine mastitis. We have also identified nisin derivatives with an enhanced ability to impair biofilm formation and to reduce the density of established biofilms of methicillin resistant S. pseudintermedius. The present study was aimed at evaluating the potential of nisin and nisin derivatives to increase the efficacy of conventional antibiotics and to assess the possibility of killing and/or eradicating biofilm-associated cells of a variety of staphylococcal targets. Growth curve-based comparisons established that combinations of derivatives nisin V + penicillin or nisin I4V + chloramphenicol had an enhanced inhibitory effect against S. aureus SA113 and S. pseudintermedius DSM21284, respectively, compared to the equivalent nisin A + antibiotic combinations or when each antimicrobial was administered alone. Furthermore, the metabolic activity of established biofilms treated with nisin V + chloramphenicol and nisin I4V + chloramphenicol combinations revealed a significant decrease in S. aureus SA113 and S. pseudintermedius DSM21284 biofilm viability, respectively, compared to the nisin A + antibiotic combinations as determined by the rapid colorimetric XTT assay. The results indicate that the activities of the nisin derivative and antibiotic combinations represent a significant improvement over that of the wild-type nisin and antibiotic combination and merit further investigation with a view to their use as anti-biofilm agents.

Antibacterial activity of nisin, oregano essential oil, EDTA, and their combination against Salmonella Enteritidis for application in mayonnaise

Salmonella Enteritidis (SE) is one of the most important serovars associated with Salmonella gastroenteritis outbreaks in Brazil. The use of natural antimicrobials can be an alternative method of SE control. The antimicrobial effect of two oregano essential oils (OEO1 and OEO2) at 0.1%, 0.2%, 0.5%, 1.0%, or 2.0%; nisin (Nisaplin®) at 0, 6.25, 12.5, or 25 ppm; ethylenediaminetetraacetic acid (EDTA) at 0.0037%, 0.0056%, 0.0075%, 0.0110%, or 0.0150%; and their combination against SE in vitro was studied to be applied in mayonnaise and Russian salad made with the same mayonnaise during storage at 8o or 30oC for 24 hours. OEO was very efficient against SE at all tested concentrations, while nisin and EDTA showed no effect against SE. Ten volatile components were identified in the two OEOs using gas chromatography coupled to mass spectrometry with electron impact ionization, with carvacrol being the major component of both samples. OEO2, containing p-cymene (15.95%) and y-terpinene (6.90%), besides carvacrol (61.66%), resulted in larger inhibition zone than the other OEO (OEO1 don’t contains p-cymene or y-terpinene). Nisin in combination with OEO1 or OEO2 had an antagonistic effect at all concentrations. The presence of nisin caused a reduction in essential oil antimicrobial activity (p≤0.05). Sensory evaluation showed that consumers prefer 0.2% OEO in mayonnaise instead of 0.5% and 1.0% concentrations. Thus, OEO only, at a concentration of 0.2%, was applied in mayonnaise against SE. The Russian salad prepared with mayonnaise plus OEO at 0.2% (wt/wt) caused a reduction of SE when compared with the salad prepared with mayonnaise without OEO. These results indicate that the use of OEO as a biopreservative (natural antimicrobial) can enhance food safety, serving as an additional barrier in helping the Good Manufacturing Practices and the Hazard Analysis Critical Control Point program, fundamental to food safety.

Modelling the combined effect of salt, sorbic acid and nisin on the probability of growth of Clostridium sporogenes in high moisture processed cheese analogue

This study focused on the application of hurdle technology to inhibit the growth of Clostridium sporogenes in high moisture, low salt, ambient shelf-stable processed cheese analogue (PCA). The effectiveness of various combinations of salt, potassium sorbate and nisin in inhibiting C. sporogenes growth was quantified using logistic regression to produce probability-of-growth models. The results of this study demonstrate that it is feasible to produce a high moisture and low salt PCA, if sufficient concentrations of potassium sorbate and nisin are added to the product. Any change in the concentration of one of the selected preservatives must be compensated for by changes in concentrations of the other two preservatives. The models developed enable the level of probability to be set to provide a required level of stringency. The models tend to be fail-safe as they were developed using an inoculum size larger than the expected contamination in processed cheese.

Effects of Bio-preservative Combined with Partially Frozen Storage on Fresh-keeping of &lt;em&gt;Argopectens irradias&lt;/em&gt; Meat

Treated with lysozyme and nisin and vacuum packed, preservation effect of the bio-preservatives on Argopectens irradias meat under partial freezing condition of -3C has been studied. During the process, Total Bacterial Count (TBC) and some physical and biochemical data had been determined. Results show that after 20 days storage at -3C, Total Volatile Basic Nitrogen (T-VBN) of the Argopectens irradias meat treated with lysozyme and nisin reached 13.28 and 13.98 mg/100 g, respectively which all approached upper control limit by China national sanitary standard, while the test datum of Argopectens irradias meat treated with combination of lysozyme and nisin reached 14.81 mg/100 g after 30 days storage which is 20 days longer than control sample. Storage time of each sample correlate to pH, TBC, T-VBN, Thiobarbituric Acid (TBA) and Ca2+-ATPase activity (p<0.05) of the test meat. Of all test samples, combination treatment with lysozyme and nisin achieved the best preservation effect.

Efficacy of a combination of nisin and p-Anisaldehyde against Listeria monocytogenes

Nisin is used as a food-safe antimicrobial agent and has been widely applied to daily food products to prevent bacterial growth. However, the practical application of nisin is limited. Although nisin inhibits the growth of Listeria monocytogenes, exposure to this agent is also a risk factor for the development of nisin resistance. p-Anisaldehyde (AS) is used in the pharmaceutical industry to manufacture antimicrobial drugs, but few studies have been reported describing the antibacterial activity of AS in the food field. Combining nisin with AS may enhance antimicrobial activity. The synergy observed in vitro was confirmed by checkerboard microdilution method, with the fractional inhibitory concentration index (FICI) values ranging from 0.125 to 0.281. When AS was combined with nisin, a strong synergistic effect was observed using the time-kill and agar-diffusion assays. SEM and LIVE/DEAD BacLight experiment results also suggest that the bactericidal mechanism of the drug combination involves cell wall lysis and membrane damage. We can use this synergy to achieve a stable antimicrobial effect to overcome antimicrobial drug resistance.

Integration of nisin into nanoparticles for application in foods

Abstract Nisin and other similar food preservatives of natural origins have seen a substantial upsurge in global interest in recent years. The use of nanotechnology to regulate and manipulate nisin for enhanced capacities in the food and nutrition sector is expanding dramatically. Nanotechnology has significant food science applications in nanoparticle delivery systems, packaging, food security, and safety. However, there have been considerable issues regarding the use of nisin in the food sector, including its uncontrolled interactions with various food components, its degradation and electrostatic repulsion. These issues potentially limit its use. Alternate strategies, including a variety of nanoparticle systems such as nanoemulsions, polymeric nanoparticles, nanofibers, and combinations of nisin with other technologies, are employed to enhance the utility of nisin in the food industry. This review highlights the recent developments and new perspectives related to the uses of nisin in the food industry. Industrial relevance This review highlights the current status of nanotechnology in the food industry. The issues concerning use of nisin by the food industry are addressed. Nisin shows enhanced efficacy in combination with other current technologies for improved food safety.

Effect of heat, nisin and ethylene diamine tetra‐acetate treatments on shelf life extension of liquid whole egg

SummaryThis study examined the effect of nisin alone or in combination with pasteurisation or ethylene diamine tetra‐acetate (EDTA), on the shelf life of unpasteurised liquid whole egg (LWE) stored at refrigerator temperature for a period of 21 days. The shelf life of samples was determined by counting total mesophilic bacteria, yeast and mould and also pH changes. It was observed that addition of 3 ppm nisin to pasteurised LWE was effective enough to extend the shelf life of this product from about 7 days to at least 21 days. However, for unpasteurised LWE nisin alone had no significant effect on shelf life while treatment of LWE with nisin in combination with EDTA showed a significant antibacterial and antifungal activity leading to extended shelf life of unpasteurised LWE from 1 to 4 days to at least 21 days.

Synergistic Antibacterial Effect of the Combination of ɛ-Polylysine and Nisin against Enterococcus faecalis

This study evaluated the antibacterial effect of the combination of ε-polylysine (ε-PL) and nisin against Enterococcus faecalis strains. The combination of ε-PL and nisin showed synergistic antibacterial activity against three Enterococcus strains. Scanning electron microscopy and a membrane permeability assay revealed that the combined treatment with ε-PL and nisin synergistically damaged the cell morphology of E. faecalis strain R612Z1 cells. Both ε-PL and nisin can dissipate the transmembrane electric potential of E. faecalis R612Z1 cells, but these peptides did not affect the transmembrane pH gradient. The combination of ε-PL and nisin can produce a high reactive oxygen species level in E. faecalis R612Z1 cells. The results indicated that the uptake of ε-PL into cells was promoted through nisin and that the combination of ε-PL and nisin could produce a high reactive oxygen species level in E. faecalis R612Z1 cells, leading to cell growth inhibition.

Effect of Nisin and Thermal Treatments on the Heat Resistance of Clostridium sporogenes Spores

The aim of this research was to evaluate the effect of thermal treatments (isothermal or nonisothermal) combined with nisin, a natural antimicrobial, on the survival and recovery of Clostridium sporogenes spores. The addition of nisin to the heating medium at concentrations up to 0.1 mg liter-1 did not reduce the heat resistance of C. sporogenes. Without a thermal treatment, nisin added at concentrations up to 0.1 mg liter-1 did not reduce the viable counts of C. sporogenes when added to the recovery medium, but inactivation of more than 4 log cycles was achieved after only 3 s at 100°C. At 100°C, the time needed to reduce viable counts by more than 3 log cycles was nine times shorter when 0.01 mg liter-1 nisin was added to the recovery medium than without it. The heat resistance values calculated under isothermal conditions were used to predict the survival in the nonisothermal experiments, and the predicted values accurately fit the experimental data. The combination of nisin with a thermal treatment can help control C. sporogenes.

Temperature and pH influence the susceptibility of Salmonella Typhimurium to nisin combined with EDTA

Lantibiotics, such as nisin, generally act on Gram-positive bacteria, whereas Gram-negative bacteria exhibit natural resistance to the action of these bacteriocins. However, Gram-negative bacteria can become susceptible to the action of these antimicrobial peptides with the use of chelating agents, such as EDTA. The activity of bacteriocins can be influenced by several factors, such as environmental conditions and chemical composition of the medium. Moreover, the susceptibility of target microorganism can be altered, due to changes in cellular composition induced by environmental conditions. In this study, we examined the combined effect of variations in temperature and pH on the susceptibility of Salmonella Typhimurium to the action of nisin associated with EDTA. The reduction in the number of viable cells of Salmonella Typhimurium varied according to the temperature and pH, and major reduction in logarithmic cycles of viable cells occurred with a temperature increase. Under low temperature and low pH, a considerable reduction in the viability was also observed after prolonged exposure to bacteriocin, regardless of the presence of EDTA. According to results, temperature and pH conditions influence on the susceptibility of Salmonella Typhimurium to combination of nisin and EDTA and thus should be considered to establish optimal conditions for nisin use.