Search

This work aimed to develop amphiphilic nanocarriers such as polymersome based diblock copolymer of Kollicoat ® IR -block-poly(ε-caprolactone) (Kollicoat ® IR-b-PCL) for potential co-delivery of Nisin (Ni) and Curcumin (CUR) for treatment of breast cancer. To generate multi-layered nanocarriers of uniform size and morphology, microfluidics was used as a new technology. In order to characterise and optimize polymersome, design of experiments (Design-Expert) software with three levels full factorial design (3-FFD) method was used. Finally, the optimized polymersome was produced with a spherical morphology, small particle size (dH < 200 nm), uniform size distribution (PDI < 0.2), and high drug loading efficiency (Ni 78 % and CUR 93 %). Furthermore, the maximum release of Ni and CUR was found to be roughly 60 % and 80 % in PBS, respectively. Cytotoxicity assays showed a slight cytotoxicity of Ni and CUR -loaded polymersome (N- Ni /CUR) towards normal cells while demonstrating inhibitory activity against cancer cells compared to the free drugs. Also, the apoptosis assays and cellular uptake confirmed the obtained results from cytotoxic analysis. In general, this study demonstrated a microfluidic approach for preparation and optimization of polymersome for co-delivery of two drugs into cancer cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

In order to develop bacteriocins, like the lantibiotic nisin A, into effective alternatives to existing antibiotics, their biophysical and physicochemical properties must first be assessed, from solubility, to susceptibility and absorption. It has been well established that formulation strategies at early drug development stages can be crucial for successful outcomes during preclinical and clinical phases of development, particularly for molecules with challenging physicochemical properties. This work elucidates the physicochemical challenges of nisin A in terms of its susceptibility to digestive enzymes like pepsin, pancreatin and proteinase K and its poor solubility at physiological pHs. Low solution concentrations, below the minimum inhibitory concentration against Staphylococcus aureus, were obtained in phosphate buffered saline (PBS, pH 7.4) and in fasted state simulated intestinal fluid (FaSSIF, pH 6.5), while higher solubilities at more acidic pH's such as in a KCl/HCl buffer (pH 2) and in fasted state simulated gastric fluid (FaSSGF, pH 1.6) are observed. Tween® 80 (0.01% v/v) significantly increased the solution concentration of nisin A in PBS (pH 7.4, 24 hr). Pancreatin doubled nisin A's solution concentration at pH 7.4 (PBS) but reduced its' inhibitory activity to ∼ 20%, and pepsin almost completely degraded nisin (after 24 hr), but retained activity at biologically relevant exposure times (∼15 min). Harnessing synergism between nisin A and either glycol chitosan or ε-poly lysine, combined with the solubilizing effect of Tween®, increased the antimicrobial activity of nisin A six fold in an in vitro oral administration model., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Numerous beneficial microbes thrive in the oral cavity where they form biofilms on dental and mucosal surfaces to get access to nutrients, and to avoid being carried away with the saliva. However, biofilm formation is also a virulence factor as it also protects pathogenic bacteria, providing them with an environment for proliferation causing oral infections. Oral hygiene relies on mechanical removal of biofilms. Some oral care products also contain antimicrobials, but effective eradication of biofilms with antimicrobials requires both a high concentration and long exposure time. In the present communication, we investigate the potential of using miniaturized drug delivery devices, known as microcontainers (MCs), to deliver the antimicrobial peptide, nisin to an oral multi-species biofilm. MCs are loaded with nisin and X-ray micro-computed tomography reveals a full release of nisin through a chitosan lid within 15 min. Chitosan-coated MCs display substantial bioadhesion to the buccal mucosa compared to non-coated MCs (68.6 ± 14.3% vs 33.8 ± 5.2%). Confocal monitoring of multi-species biofilms reveals antibacterial effects of nisin-loaded chitosan-coated MCs with a faster onset (after 3 h) compared to solution-based delivery (after 9 h). Our study shows the potential of using MCs for treatment of multi-species oral biofilms and is encouraging for further design of drug delivery devices to treat oral diseases., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Salmonella enterica is a pathogen that induces self-limiting gastroenteritis and is of worldwide concern. Nisin, an antimicrobial peptide, has emerged as an alternative for the control of microbial growth but its effect on the virulence of pathogenic bacteria is not yet well-explored. This work aimed to evaluate the virulence of S. enterica in the presence of sub-inhibitory nisin using the experimental model Galleria mellonella. Sub-inhibitory concentrations of nisin of 11.72 and 46.88 μM did not affect the cellular viability of S. enterica but promoted changes in gene expression within 1 h of treatment, with increases of up to 3-fold of pagC, 1.8-fold of invA and 2.3-fold of invF. Larvae of G. mellonella inoculated with S. enterica combined with nisin at 46.88 μM presented mortality, and TL 50 noticeably increased to 50% and 80% at 24 and 48 h post-infection, respectively. Defence responses, such as melanisation, nodulation, pseudopodia, immune response, and expression of defence proteins of the larvae G. mellonella were enhanced when the treatments with S. enterica were combined with 11.72 or 46.88 μM nisin. These results show an increase in virulence of S. enterica by sub-MIC concentration of nisin that needs to be explored., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

The current study investigated Listeria monocytogenes inactivation using mild heat with elevated hydrostatic pressure and nisin under buffered condition. A four-strain pathogen mixture was exposed to 0 (control) and up to 9 min of (1) 4 °C elevated pressure; (2) 4 °C elevated pressure and nisin; (3) 4 °C nisin; (4) heat at 40 °C; (5) 40 °C elevated pressure; (6) 40 °C elevated pressure and nisin; and (7) 40 °C nisin. Elevated hydrostatic pressure at 400 MPa (Hub880 Explorer, Pressure BioScience Inc., Easton, MA, USA) and nisin concentration of 5000 IU/mL were used in the trials. Analyses of variance were conducted, followed by Dunnett's- and Tukey-adjusted means separations. Under conditions of these experiments, nisin augmented ( p < 0.05) decontamination efficacy of 40 °C heat and elevated hydrostatic pressure treatments, particularly at treatment interval of 3 min. This synergism with nisin faded away ( p ≥ 0.05) as the treatment time for thermal, high-pressure, and thermal-assisted pressure processing increased. The results of our study, thus, exhibit that practitioners and stakeholders of pressure-based technologies could benefit from synergism of mild heat and nisin for short-term, high-pressure pasteurization treatments to achieve microbial safety and economic feasibility comparable to traditional heat-treated products., Competing Interests: Authors of this publication declare no conflict of interest associated with publication of this research project. Funding agencies of this project did not have a role for designing the study, for collection, statistical analyses, data interpretation, writing the manuscript, and decision to publish the results. Thus, content of the publication does not necessarily reflect views of funding institutions.

Chemotherapy, a gold standard for treating most of the cancers, involves drastic side-effects and multidrug resistance. An attractive alternative is development of combination therapy employing antimicrobial peptides with chemotherapeutic drugs. In vivo studies: Anti-cancer therapeutic efficacy of 5-fluororuacil (5-FU) in conjunction with nisin (50 mg/kg body weight) was evaluated against murine skin cancer, in terms of tumor biostatistics, histopathology, electron microscopy, infrared spectroscopy and transcriptional studies. In vitro studies: Dose and time dependent cytotoxicity of agents were assessed against A431 cell line using MTT assay, LDH assay and acridine orange/ethidium bromide dual staining. Significant percentage decrease(s) in mean tumor volume and tumor burden were observed in nisin+ 5-FU combination treated groups as compared to alone treated groups. Histoarchitecture of treated skins demonstrated restoration towards normal skin tissue (being highest in the combination group). Modulation of apoptotic, angiogenic and proliferative genes were observed in treated groups. IC 50 of combination was found to be 2 μg/ml as compared to nisin alone (32μg/ml) and 5-FU alone (16μg/ml) with combination index of 0.188. Dual staining showed that rate of induction of apoptosis was higher in the combination group as compared to single agents. Nisin and 5-FU in combination were found to be synergistic both in vivo and in vitro., (Copyright © 2019. Published by Elsevier Inc.)

Background: Diabetic foot infections (DFIs) are a frequent complication of Diabetes mellitus and a major cause of nontraumatic limb amputations. The Gram-positive bacterium Staphylococcus aureus, known for its resilient biofilms and antibiotic resistant profile, is the most frequent DFI pathogen. It is urgent to develop innovative treatments for these infections, being the antimicrobial peptide (AMP) nisin a potential candidate. We have previously proposed the use of a guar gum biogel as a delivery system for nisin. Here, we evaluated the potential of the nisin-biogel to enhance the efficacy of conventional antibiotics and antiseptics against DFIs S. aureus clinical isolates., Methods: A collection of 23 S. aureus strains isolated from DFI patients, including multidrug- and methicillin-resistant strains, was used. The antimicrobial activity of the nisin-biogel was tested alone and in different combinations with the antiseptic chlorhexidine and the antibiotics clindamycin, gentamicin and vancomycin. Isolates' in vitro susceptibility to the different protocols was assessed using broth microdilution methods in order to determine their ability to inhibit and/or eradicate established S. aureus biofilms. Antimicrobials were added to the 96-well plates every 8 h to simulate a typical DFI treatment protocol. Statistical analysis was conducted using RCBD ANOVA in SPSS., Results: The nisin-biogel showed a high antibacterial activity against biofilms formed by DFI S. aureus. The combined protocol using nisin-biogel and chlorhexidine presented the highest efficacy in biofilm formation inhibition, significantly higher (p<0.05) than the ones presented by the antibiotics-based protocols tested. Regarding biofilm eradication, there were no significant differences (p>0.05) between the activity of the combination nisin-biogel plus chlorhexidine and the conventional antibiotic-based protocols., Conclusions: Results provide a valuable contribution for the development of complementary strategies to conventional antibiotics protocols. A combined protocol including chlorhexidine and nisin-biogel could be potentially applied in medical centres, contributing for the reduction of antibiotic administration, selection pressure on DFI pathogens and resistance strains dissemination., Competing Interests: The authors declare that research was conducted in the absence of any commercial or financial relationships that could be considered as a potential conflict of interest.

Traditional broad-spectrum antibacterial agents are limited by high toxicity and the destruction to the bacterial flora balance in the wound site. Herein, we propose an opinion that one or several especial antibacterial peptides are adopted to kill the target bacteria in order to precisely manage the bacteria infected chronic wound under the premise of biobalance, and specially employ nisin to treat S. aureus infected chronic wound as model with positive effects. The results showed that without cytotoxicity to the normal cells, only 25 ppm nisin could contrapuntally kill S. aureus and have little inhibitory to other bacteria. Mechanism of antibacterial selectivity indicated the superior biomolecular interaction between nisin and S. aureus compared with E. coli and normal cells. Furthermore, nisin significantly accelerated the healing process of S. aureus-infected rabbit full thickness burn wound, but had no effect on the E. coli-infected wound as a comparison. Therefore, it has been demonstrated that special peptides with antibacterial selectivity can be adopted to precise management for bacteria infected chronic wound under good biobalance.

Nisin, a lantibiotic bacteriocin, has been used for years as a natural food preservative. In addition to its antimicrobial activity, nisin also shows immunomodulatory properties, and the nisin-producing Lactococcus lactis strain has been successfully tested as a probiotic in weaned piglets. However, the impact of nisin on porcine immune cells has not yet been explored. The objective of the present study was to examine the in vitro immunomodulatory effect of nisin on porcine peripheral blood leucocytes. The whole heparinized blood samples or freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with different nisin concentrations (0, 1.56, 3.125, 6.25, 12.5, 25 or 50 µg/ml) for 1, 24, 48 or 72 hr. Escherichia coli bacteria were used to stimulate blood phagocytes, while concanavalin A and lipopolysaccharide from E. coli were used as mitogens. Control cells remained unstimulated. MTT colorimetric assay was used to evaluate PBMCs viability and mitogenic response. Phagocyte activity and T-cell proliferation were measured by flow cytometry. Flow cytometer was also used for immunophenotyping of T cells. Cytokine levels in the culture media were determined using commercial immunoassay (ELISA) kits. The highest concentration of nisin exhibited proliferative activity (p ˂ 0.05), stimulated interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) production (both at p ˂ 0.001), and increased the percentage of CD4 + CD8 + T cells (p ˂ 0.001) among unstimulated leucocytes. After cell stimulation, however, the highest nisin concentration showed antiproliferative activity (p ˂ 0.05), decreased phagocytic functions (p ˂ 0.05) and inhibited the synthesis of IL-6 (time- and concentration-dependent effect). As a typical bacterial product, nisin had a stronger impact on innate immune cells, and its effect on T cells was likely a consequence of the modulation of the activity of antigen-presenting cells. Nisin may be a good candidate as an immunomodulator in pig breeding., (© 2019 Blackwell Verlag GmbH.)

Background: Periodontal disease (PD) is caused by the development of a microbial biofilm (dental plaque) in the periodontium, affecting approximately 80% of dogs. Several bacterial species present in the canine oral cavity can be implicated in the development of this disease, including Enterococcus spp. To decrease antibiotic administration, a possible control strategy for dog's enterococcal PD may involve the use of the antimicrobial peptide (AMP) nisin. Nisin's inhibitory activity was evaluated against a collection of previously characterized enterococci obtained from the oral cavity of dogs with PD (n = 20), as well as the potential of a guar-gum gel and a veterinary toothpaste as topical delivery systems for this AMP. The Minimum Inhibitory (MIC) and Bactericidal Concentrations (MBC) and the Minimum Biofilm Eradication (MBEC) and Inhibitory Concentrations (MBIC) were determined for nisin and for the supplemented guar-gum gel. For the supplemented veterinary toothpaste an agar-well diffusion assay was used to evaluate its inhibitory potential., Results: Nisin was effective against all isolates. Independently of being or not incorporated in the guar-gum gel, its inhibitory activity on biofilms was higher, with MBIC (12.46 ± 5.16 and 13.60 ± 4.31 μg/mL, respectively) and MBEC values (21.87 ± 11.33 and 42.34 ± 16.61 μg/mL) being lower than MIC (24.61 ± 4.64 and 14.90 ± 4.10 μg/mL) and MBC (63.09 ± 13.22 and 66.63 ± 19.55 μg/mL) values. The supplemented toothpaste was also effective, showing inhibitory activity against 95% of the isolates., Conclusions: The inhibitory ability of nisin when incorporated in the two delivery systems was maintained or increased, demonstrating the potential of these supplemented vehicles to be applied to PD control in dogs.

Myocardial infarction (MI) is the most prevalent cause of cardiovascular death. A possible way of preventing MI maybe by dietary supplements. The present study was thus designed to ascertain the cardio-protective effect of a formulated curcumin and nisin based poly lactic acid nanoparticle (CurNisNp) on isoproterenol (ISO) induced MI in guinea pigs. Animals were pretreated for 7 days as follows; Groups A and B animals were given 0.5 mL/kg of normal saline, group C metoprolol (2 mg/kg), groups D and E CurNisNp 10 and 21 mg/kg respectively (n = 5). MI was induced on the 7 th day in groups B-E animals. On the 9 th day electrocardiogram (ECG) was recorded, blood samples and tissue biopsies were collected for analyses. Toxicity studies on CurNisNp were carried out. MI induction caused atrial fibrillation which was prevented by pretreatment of metoprolol or CurNisNp. MI induction was also associated with increased expressions of cardiac troponin I (CTnI) and kidney injury molecule-1 (KIM-1) which were significantly reduced in guinea pig's pretreated with metoprolol or CurNisNp (P < 0.05). The LC 50 of CurNisNp was 3258.2 μg/mL. This study demonstrated that the formulated curcumin-nisin based nanoparticle confers a significant level of cardio-protection in the guinea pig and is nontoxic.

Colorectal cancer is the third most common cause of cancer-related death in the world which genetic and environmental agents are responsible for cancer. When cells detach from the tumor and invade surrounding tissues, the tumor is malignant and may form secondary tumors at other locations in a process called metastasis. Probiotics are the largest group of inhabitation bacteria in the colon. Gut microbiota has a central role in prevented the risk colon cancer. Probiotics are beneficial microorganisms, like Lactic acid bacteria and Lactobacilli bacteria which are using in the dairy industry. Probiotics nisin are having the most important category of safe usage. In this study LS180, SW48, HT29 and Caco2 was cultured and treated with different dose of nisin. Cell proliferation was assayed with MTT. The expression of CEA, CEAM6 and MMP2F genes was analyzed with Real-time PCR. Protein expression of CEA was evacuated with ELISA. Our result was shown that the 40-50 IU/mL nisin could suppress proliferation of LS180. Cell proliferation of SW48, HT29, Caco2 cells was decreased in 250-350 IU/mL concentration of nisin. The gene expression of CEA, CEAM6, MMP2F was significantly down-regulated with nisin treatment (p < 0.001, p < 0.01). Also, after cells treated with nisin, CEA protein expression was down regulated (p < 0.01). In conclusion, nisin could suppressed metastatic process via down-regulation of CEA, CEAM6, MMP2F, MMP9F genes. We suggested the new treatment strategies beyond Probiotics, which play a role in the prevention local tumor invasion, metastasis and recurrence., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Reverse micelles (RMs) as nanocarriers of nisin were optimized for the highest water and bacteriocin content. RMs formulated with either refined olive oil or sunflower oil, distilled monoglycerides, ethanol, and water were effectively designed. Structural characterization of the RMs was assessed using Electron Paramagnetic Resonance Spectroscopy and Small Angle X-ray Scattering in the presence and absence of nisin. No conformational changes occurred in the presence of nisin for the nanocarriers. To assess efficacy of the loaded systems, their antimicrobial activity against Staphylococcus aureus and Listeria monocytogenes was tested in lettuce leaves and minced meat, respectively. Antimicrobial activity was evident in both cases. Interestingly, a synergistic antimicrobial effect was observed in lettuce leaves and to a lesser extent in minced meat between nisin and some of the nanocarriers' constituents (probably ethanol). Our findings suggest complex interactions that take place when RMs are applied in different food matrices., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Japanese cedar pollinosis is a seasonal allergic disease caused by two major pollen allergens: Cry j 1 and Cry j 2 antigens. To develop an oral vaccine to treat pollinosis, we constructed recombinant Lactococcus lactis harboring the gene encoding fused T cell epitopes from the Cry j 1 and Cry j 2 antigens. The recombinant T cell epitope peptide was designed to contain the fused cholera toxin B subunit as an adjuvant and a FLAG tag at the C-terminus. An expression plasmid was constructed by inserting the T cell epitope peptide gene into the multiple cloning sites of plasmid pNZ8148, an Escherichia coli-L. lactis shuttle vector. The constructed plasmid was transformed into L. lactis NZ9000 for expression induced by nisin, an antibacterial peptide from L. lactis. The expression of the epitope peptide was induced with 10-40 ng/mL nisin, and the expressed T cell epitope peptide was detected by western blot analysis using an anti-FLAG antibody and an antibody against the T cell epitopes. The concentration of the epitope peptide was estimated to be ~ 22 mg/L of culture in the presence of 40 ng/mL nisin, although it varied depending on the nisin concentration, the culture time, and the bacterial concentration when nisin was added. The expression of the recombinant epitope peptide in L. lactis, an organism generally recognized as safe, as demonstrated in this study, may contribute to the development of an oral vaccine for the treatment of pollinosis.

Nisin is a polycyclic peptide containing 34 amino acids produced by Lactococcus lactis during fermentation. Recently, researchers considered nisin as an anticancer peptide. Herein, the authors aim to evaluate the nisin effects on the apoptosis stimulation in the colon cancer cell line. The SW480 cells were exposed to discrepant concentrations of nisin and the cell viability as well as the expression of bcl-2 and bax genes and proteins were surveyed by the MTT assay, Real-Time PCR and western blotting method, respectively. Furthermore, the Ethidium bromide/Acridine orange staining was performed to visualize apoptotic cells. 4000, 3000, 2500 and 2000 μg/ml of nisin led to significant anti-proliferative impact and augmentation apoptotic index (bax/bcl-2 ratio) both at mRNA and protein levels (p < 0.05). Furthermore, the apoptotic impacts were demonstrated after Ethidium bromide/Acridine orange (EB/AO) staining to have a dose dependent manner. Our outcome suggested that nisin could induce apoptosis via intrinsic pathways and lead to cancerous cell death., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Introduction: Nisin, a broad-spectrum bacteriocin, has recently been highlighted for its biomedical applications. To date, no studies have examined the antimicrobial and antibiofilm properties of high-purity (>95%) nisin (nisin ZP) on Enterococcus faecalis and biofilms formed by this species. We hypothesize that nisin can inhibit E. faecalis and reduce biofilm biomass, and combinations of nisin and sodium hypochlorite (NaOCl) will enhance the antibiofilm properties against E. faecalis biofilms., Methods: Using broth cultures, disc diffusion assays, and biofilm assays, we examined the effects of nisin on various E. faecalis growth parameters and biofilm properties (biovolume, thickness, and roughness). Confocal microscopy was used in conjunction with Imaris and Comstat2 software (Kongens Lyngby, Copenhagen, Denmark) to measure and analyze the biofilm properties., Results: Nisin significantly decreased the growth of planktonic E. faecalis dose dependently. The minimum inhibitory concentrations against E. faecalis strains OG-1 and ATCC 29212 were 15 and 50 μg/mL, and the minimum bactericidal concentrations were 150 and 200 μg/mL, respectively. A reduction in biofilm biovolume and thickness was observed for biofilms treated with nisin at ≥10 μg/mL for 10 minutes. In addition, the combination of nisin with low doses of NaOCl enhanced the antibiofilm properties of both antimicrobial agents., Conclusions: Nisin alone or in combination with low concentrations of NaOCl reduces the planktonic growth of E. faecalis and disrupts E. faecalis biofilm structure. Our results suggest that nisin has potential as an adjunctive endodontic therapeutic agent and as an alternative to conventional NaOCl irrigation., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This work is focused on preparation of novel porous type of core-shell-structured microparticles based on polylactide (shell) and poly(vinyl alcohol) cross-linked with glutaric acid (GA) (core) prepared by water-in-oil-in-water solvent evaporation technique. The microparticle systems were used as delivery systems for immobilisation of model antibacterial agent - nisin. The effect of cross-linking and the initial amount of nisin on their morphology was investigated using scanning electron microscopy, BET analysis, zeta potential measurement and Fourier transform infra-red spectroscopy. Encapsulation efficiency and release profile of nisin from the microparticles were studied by high performance liquid chromatography. Antibacterial activity of the prepared systems was tested by dilution and spread plate technique. Results showed the microparticles in the size range of 9-16 μm in diameter with spherical multi-hollow core-shell structure. The presence of cross-linking agent GA influences the release profile of the peptide and has synergistic effect on Listeria monocytogenes growth reduction.

Electrospun membranes encapsulating nisin in the core of multi-layer coaxial fibers, with a hydrophobic PCL intermediate layer and a hygroscopic cellulose acetate sheath, have been demonstrated to provide long-term antimicrobial activity combined with a hygroscopic outer layer. Antimicrobial performance has been evaluated using modified versions of the antimicrobial textile test AATCC 100 and AATCC 147 against Staphylococcus aureus. The AATCC 147 tests indicate that antimicrobial activity persists up to 7days. The quantitative analysis from the AATCC 100 test indicates that tri-layer coaxial ("triaxial") electrospun fiber membranes provide >99.99% bacteria kill (4logkill) for up to five days. This indicates that the nisin-incorporated triaxial fibers have excellent biocidal activities for up to 5days and then provide biostatic activity for 2 or more days. Compared with other types of electrospun membranes, such as core-sheath coaxial ("coaxial") and single homogenous fibers, triaxial fiber membranes provided more robust and more sustained antimicrobial activity. Single fibers with nisin showed relatively weak activity and only for one day. Coaxial fiber membranes exhibited antimicrobial activity for a long period, but their biocidal activity was much weaker than that of triaxial fiber membranes, and only exhibited >99% bacteria kill (2logkill) after 1day of exposure., Statement of Significance: The increase in drug resistant pathogens has driven the need for alternative treatments that are effective against resistant bacteria and do not contribute to drug resistance. Nisin is an excellent model bacteriocin for antimicrobials because of its size and mode of action, and has been extensively used as FDA-approved food preservatives without any problematic resistance growth in bacteria during past decades. Nisin-containing fibers have been previously reported using conventional electrospinning but sustained antimicrobial effect has not been obtained. Here, we report the encapsulation of nisin into a multi-layered nanofiber construct using triaxial electrospinning in order to obtain a long-term antimicrobial activity. This will be highly beneficial in many applications, such as protective textiles, food packaging and cancer therapy., (Copyright © 2017 Acta Materialia Inc. All rights reserved.)

This study evaluated the effect of a Staphylococcus aureus bacterin and nisin on bovine subclinical mastitis. A total of 75 Holstein subclinically mastitic cows were randomly allocated to three groups with 25 cows per group. In group I, an intramammary infusion of nisin Z at a dose of 2.50×10 6 IU was carried out once daily for three days, and an autogenous S. aureus bacterin was inoculated into the supramammary lymph node one week before and one week after nisin treatment. In group II, nisin was administered in the same way as in group I, but no bacterin was inoculated. Group III received no treatment and served as a control. Milk was aseptically sampled from the affected quarters before and 2, 4, and 6 weeks after treatment, for bacteriological examination and analyses of N-acetyl-β-D-glucosaminidase (NAGase) activity, somatic cell count (SCC), and milk protein and fat contents. Results indicated that, compared to the nisin-treated group, nisin-bacterin treatment significantly reduced intramammary S. aureus infections, reduced the number of quarters with milk SCCs of more than 5×10 5 cells/ml, and increased the protein and fat contents of the milk. Therefore, nisin-bacterin therapy is suggested when subclinical mastitis occurs in lactating cows.

The aim of this work was the development and characterization of nisin-loaded nanoparticles and the evaluation of its potential antifungal activity. Candidiasis is a fungal infection caused by Candida sp. considered as one of the major public health problem currently. The discovery of antifungal agents that present a reduced or null resistance of Candida sp. and the development of more efficient drug release mechanisms are necessary for the improvement of candidiasis treatment. Nisin, a bacteriocin commercially available for more than 50 years, exhibits antibacterial action in food products with potential antifungal activity. Among several alternatives used to modulate antifungal activity of bacteriocins, polymeric nanoparticles have received great attention due to an effective drug release control and reduction of therapeutic dose, besides the minimization of adverse effects by the preferential accumulation in specific tissues. The nisin nanoparticles were prepared by double emulsification and solvent evaporation methods. Nanoparticles were characterized by dynamic light scattering, zeta potential, Fourier transform infrared, X-ray diffraction, differential scanning calorimetry, and scanning electron microscopy. Antifungal activity was accessed by pour plate method and cell counting using Candida albicans strains. The in vitro release profile and in vitro permeation studies were performed using dialysis bag method and pig vaginal mucosa in Franz diffusion cell, respectively. The results revealed nisin nanoparticles (300 nm) with spherical shape and high loading efficiency (93.88 ± 3.26%). In vitro test results suggest a promising application of these nanosystems as a prophylactic agent in recurrent vulvovaginal candidiasis and other gynecological diseases.

Nisin is a bacteriocin produced by a group of Gram-positive bacteria that belongs to Lactococcus and Streptococcus species. Nisin is classified as a Type A (I) lantibiotic that is synthesized from mRNA and the translated peptide contains several unusual amino acids due to post-translational modifications. Over the past few decades, nisin has been used widely as a food biopreservative. Since then, many natural and genetically modified variants of nisin have been identified and studied for their unique antimicrobial properties. Nisin is FDA approved and generally regarded as a safe peptide with recognized potential for clinical use. Over the past two decades the application of nisin has been extended to biomedical fields. Studies have reported that nisin can prevent the growth of drug-resistant bacterial strains, such as methicillin-resistant Staphylococcus aureus, Streptococcus pneumoniae, Enterococci and Clostridium difficile. Nisin has now been shown to have antimicrobial activity against both Gram-positive and Gram-negative disease-associated pathogens. Nisin has been reported to have anti-biofilm properties and can work synergistically in combination with conventional therapeutic drugs. In addition, like host-defence peptides, nisin may activate the adaptive immune response and have an immunomodulatory role. Increasing evidence indicates that nisin can influence the growth of tumours and exhibit selective cytotoxicity towards cancer cells. Collectively, the application of nisin has advanced beyond its role as a food biopreservative. Thus, this review will describe and compare studies on nisin and provide insight into its future biomedical applications., (© 2015 The Authors. published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

We aimed to investigate the antibacterial activities of carvacrol, thymol, eugenol, cinnamaldehyde, and lantibiotic nisin against standard bacterial strains of the milk pathogens Staphylococcus aureus ATCC 25923 and Listeria monocytogenes ATCC 15313 in cow milk. The minimum inhibitory concentrations (MIC) of these substances were recorded. The synergistic effects were also assessed in culture medium (time kill curve) and in a food model (cow milk) during the storage period (4 °C for 6 d) after inoculation with S. aureus and L. monocytogenes individually by combining nisin and the phenolic compounds at proportions of 1/4 + 1/4 the MIC (determined in a previous in vitro assay) in the culture medium and 1/4 + 1/4 of MIC in the food model. Inhibitory activities of nisin and the tested compounds, as well as synergism in the combinations, were found against both bacteria assayed. Bacteriostatic effects were found with all combinations and a significant difference in L. monocytogenes reduction was found compared with the control assays. Thus, the antibacterial activity of nisin combined with phenolic compounds was confirmed against these pathogenic bacteria that are important in the milk industry, or more broadly in food science, with potential applications for milk preservation., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In view of the emergence of multidrug-resistant cancer cells, there is a need for therapeutic alternatives. Keeping this in mind, the present study was aimed at evaluating the synergism between nisin (an antimicrobial peptide) and doxorubicin (DOX) against DMBA-induced skin carcinogenesis. The possible tumoricidal activity of the combination was evaluated in terms of animal bioassay observations, changes in hisotological architecture of skin tissues, in situ apoptosis assay (TUNEL assay) and in terms of oxidant and antioxidant status of the skin tissues. In vivo additive effect of the combination was evidenced by larger decreases in mean tumour burden and tumour volume in mice treated with the combination than those treated with the drugs alone. Histological observations indicated that nisin-DOX therapy causes chromatin condensation and marginalisation of nuclear material in skin tissues of treated mice which correlated well with the results of TUNEL assay wherein a marked increase in the rate of apoptosis was revealed in tissues treated with the combination. A slightly increased oxidative stress in response to the adjunct therapy as compared to dox-alone-treated group was revealed by levels of lipid peroxidation (LPO) and nitrite generation in skin tissue-treated mice. An almost similar marginal enhancement in superoxide dismutase levels corresponding with a decrease in catalase activity could also be observed in nisin + DOX-treated groups as compared to nisin and dox-alone-treated groups. These results point towards the possible use of nisin as an adjunct to doxorubicin may help in developing alternate strategies to combat currently developing drug resistance in cancer cells.

Antibiotic resistance and the shortage of novel antimicrobials are among the biggest challenges facing society. One of the major factors contributing to resistance is the use of frontline clinical antibiotics in veterinary practice. In order to properly manage dwindling antibiotic resources, we must identify antimicrobials that are specifically targeted to veterinary applications. Nisin is a member of the lantibiotic family of antimicrobial peptides that exhibit potent antibacterial activity against many gram-positive bacteria, including human and animal pathogens such as Staphylococcus, Bacillus, Listeria, and Clostridium. Although not currently used in human medicine, nisin is already employed commercially as an anti-mastitis product in the veterinary field. 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 (MRSA) and vancomycin-resistant enterococci (VRE) and also against staphylococci and streptococci associated with bovine mastitis. However, newly emerging pathogens such as methicillin resistant Staphylococcus pseudintermedius (MRSP) pose a significant threat in terms of veterinary health and as a reservoir for antibiotic resistance determinants. In this study we created a nisin derivative with enhanced antimicrobial activity against S. pseudintermedius. In addition, the novel nisin derivative exhibits an enhanced ability to impair biofilm formation and to reduce the density of established biofilms. The activities of this peptide represent a significant improvement over that of the wild-type nisin peptide and merit further investigation with a view to their use to treat S. pseudintermedius infections.

Objectives: To evaluate the combined effect of a mixture of tetracycline, acid, and detergent (MTAD) and Nisin against Enterococcus faecalis (E. faecalis) and Actinomyces viscosus (A. viscosus) biofilms., Methods: This study was conducted between June and December 2013 in collaboration with Dental Caries Research Chair, College of Dentistry, King Saud University, Riyadh, Saudi Arabia. Single-species biofilms (n=9/species/observation period) were generated on membrane filter discs and subjected to 5, 10, or 15 minute incubation with MTADN (MTAD with 3% Nisin), 5.25% sodium hypochlorite (NaOCl), or normal saline. The colony forming units were counted using the Dark field colony counter., Results: A 100% bactericidal effect of 5.25% NaOCl was noted during the 3 observation periods; a significant reduction (p=0.000) in mean survival rates of E. faecalis (77.3+13.6) and A. viscosus (39.6+12.6) was noted after 5 minutes exposure to MTADN compared with normal saline (78000000+5291503) declining to almost no growth after 10 and 15 minutes. The survival rates of the E. faecalis and A. viscosus biofilm were no different after treatment with MTADN and 5.25% NaOCl at the 3 observation periods (p=1.000)., Conclusion: A combination of MTAD and Nisin was as effective as NaOCl against E. faecalis and A. viscosus biofilms.

Electrospinning technique was used to fabricate the model drug Nisin loaded phosphorylated soybean protein isolate/poly(l-lactic acid)/zirconium dioxide (Nisin-PSPI/PLLA/ZrO2) nanofibrous membranes. The average diameter of drug carrier PSPI/PLLA/ZrO2 nanofibers increased with the increase of content PSPI and some spindle-shape beads appeared when PSPI content reached 25 wt%. The loading dosage of Nisin caused no significant changes in the size and morphology of nanofibers when Nisin content was below 9 wt%. There existed hydrogen and Zr-O-C bonds among PSPI, PLLA and ZrO2 units, and the crystalline of PLLA matrix decreased owning to the introducing of PSPI and ZrO2 units. Moreover, the water absorption capability and degradation rate of PSPI/PLLA/ZrO2 nanofibrous membranes increased with increasing PSPI content. The antimicrobial activity and release experimental results showed that Nisin-PSPI/PLLA/ZrO2 nanofibrous membranes displayed well controlled release and better antimicrobial activity against Staphylococcus aureus (S. aureus), and the Nisin release from the medicated nanofibers could be described by Fickian diffusion model. The Nisin-PSPI/PLLA/ZrO2 nanofibrous membranes may have potential as a new nanofibrous membrane in drug delivery, food active packaging and wound dressing., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Although the antimicrobial peptide nisin has been extensively studied in the food industry for decades, its application in the oral cavity remains to develop and evaluate its feasibility in treating oral common diseases. Nisin is an odorless, colorless, tasteless substance with low toxicity and with antibacterial activities against Gram-positive bacteria. These biologic properties may establish its use in promising products for oral diseases. This article summarizes the antibacterial efficiency of nisin against pathogenic bacteria related to dental caries and root canal infection and discusses the combination of nisin and common oral drugs., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Hydrophilic nisin-loaded hydrophobic poly (lactic acid) (PLA) particles with controlled size and shape were successfully produced utilizing a one-step single emulsification method. Preliminary shear stress and temperature tests showed that there was no significant loss in the nisin inhibition activity during this process. PLA/nisin composite particles were prepared into solid nanocomposite spheres (50-200 nm) or hollow microcomposite spheres (1-5 μm) under the operative conditions developed in our previous study, in which the hydrophilic nisin in the aqueous phase solution could be entrapped in the hydrophobic polymer in the emulsification process generating either single or multiple emulsions. The incorporation of nisin in PLA had little effect on key processing conditions, which allows the dynamic control of the morphology and property of resulting particles. Microscopic and surface analyses suggested that nisin was dispersed uniformly inside the polymer matrix and adsorbed on the particle surface. The encapsulation amount and efficiency were enhanced with the increase in nisin loading in the aqueous solution. Unique reversible control of particle size and shape by this process was successfully applied in the nisin encapsulation. Alternating temperature in the repeating emulsification steps improved the encapsulation efficiency while generated particles in desired size and shape., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Bacteriocins are antimicrobial peptides produced by a variety of bacteria. These peptides can act as antibiotic synergists or alternatives to enhance the therapeutic effects of current infection treatments and decrease the prevalence of resistant strains. Two bacteriocins, namely nisin and pediocin PA-1, are currently being used by the food industry; however, the introduction of these and others into the biomedical industry, and further development of food applications, have been challenged by the slow development of reliable delivery systems. For bacteriocins, these systems rely on novel and pre-existing technologies. Many essential variables need to be accounted for to formulate successful delivery methods. In this review, documented and potential bacteriocin delivery systems are examined, with special attention paid to how those systems are being implemented in the food and medical industries.

The aim of this study was to evaluate the combined effect of a previous mild heat treatment (15 min at 55 ) with the use of antimicrobials, nisin and carvacrol, on the growth of Salmonella enteritidis and Salmonella senftenberg. Natural antimicrobials, alone or combined with a previous mild heat treatment, affected the growth of these two serovars in Tryptone Soy Broth at 37 . Increasing concentrations of carvacrol had a significant effect on both growth rate and lag phase duration of both strains. The time to reach stationary phase was almost doubled in the case of S. enteritidis when a concentration of 0.77 mM in carvacrol was added. For S. senftenberg the effect was smaller. The effect of nisin and of heat, applied individually, was lower for both microorganisms. A combination of 1.2 µM nisin with 0.77 mM carvacrol significantly delayed the growth of heat treated cells, compared to the control without antimicrobials, showing additive effects.

Innate immunity is triggered by a variety of bacterial molecules, resulting in both protective and potentially harmful pro-inflammatory responses. Further, innate immunity also provides a mechanism for the maintenance of homeostasis between the host immune system and symbiotic or non-pathogenic microorganisms. However, the bacterial factors that mediate these protective effects have been incompletely defined. Here, it was demonstrated that the lantiobiotic nisin Z is able to modulate host immune responses and mediate protective host immunity. Nisin Z induced the secretion of the chemokines MCP-1, IL-8 and Gro-α, and significantly reduced TNF-α induction in response to bacterial LPS in human PBMC. The results correlated with the ability of nisin Z to confer protection against both the Gram-positive organism Staphylococcus aureus, and the Gram-negatives Salmonella enterica sv. Typhimurium and Escherichia coli in murine challenge models. Mechanistic studies revealed that nisin Z modulates host immunity through similar mechanisms as natural host defense peptides, engaging multiple signal transduction pathways and growth factor receptors. The results presented herein demonstrate that, in addition to nisin Z, other bacterial cationic peptides and, in particular, the lantibiotics, could represent a new class of secreted bacterial molecule with immunomodulatory activities.

Introduction: Enterococcus faecalis is frequently isolated from both secondary and persistent root canal infections. MTAD is an effective intracanal irrigant, but its bactericidal activity requires improvement. The goal of this study was to investigate whether nisin, an antibacterial peptide, improves the antibacterial and antibiofilm activities of MTAD against E. faecalis clinical isolates., Methods: Ten E. faecalis strains were isolated from root canals with persistent periapical lesions. The antibacterial activity of MTAD, MTADN (nisin in combination with doxycycline), and MTAN (nisin in place of doxycycline) against E. faecalis isolates was evaluated using minimal inhibitory concentration and minimal bactericidal concentration assays. Scanning electron microscopy was used to observe E. faecalis biofilms on the chamber surface of the root canal, and confocal laser scanning microscopy was used to show the antibiofilm activity of the 3 irrigants., Results: Ten E. faecalis strains were successfully isolated from clinical samples by selective medium, colony morphology, oxygen tolerance, bile resistance, Gram staining characteristics, and polymerase chain reaction identification. The antimicrobials MTAD, MTADN, and MTAN had different antibacterial activities against the 10 E. faecalis isolates, with MTADN showing superior antibacterial efficacy compared with MTAD. Strain D had the greatest sensitivity to MTAD and MTADN, whereas strain E was the most resistant to these compounds. Strain E was able to form a mature biofilm with complex architecture. Importantly, confocal laser scanning microscopic images showed that MTADN was able to kill completely E. faecalis isolates grown as a biofilm, whereas MTAD was unable to do so., Conclusions: MTADN effectively inhibited both the growth of E. faecalis root canal isolates and these isolates' biofilms. These results are encouraging and suggest that MTADN has a considerable potential for use as an effective irrigant., (Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Background: Lantibiotics are post-translationally modified antimicrobial peptides, of which nisin A is the most extensively studied example. Bioengineering of nisin A has resulted in the generation of derivatives with increased in vitro potency against Gram-positive bacteria. Of these, nisin V (containing a Met21Val change) is noteworthy by virtue of exhibiting enhanced antimicrobial efficacy against a wide range of clinical and food-borne pathogens, including Listeria monocytogenes. However, this increased potency has not been tested in vivo., Results: Here we address this issue by assessing the ability of nisin A and nisin V to control a bioluminescent strain of Listeria monocytogenes EGDe in a murine infection model.More specifically, Balb/c mice were infected via the intraperitoneal route at a dose of 1 × 10(5) cfu/animal and subsequently treated intraperitoneally with either nisin V, nisin A or a PBS control. Bioimaging of the mice was carried out on day 3 of the trial. Animals were then sacrificed and levels of infection were quantified in the liver and spleen., Conclusion: This analysis revealed that nisin V was more effective than Nisin A with respect to controlling infection and therefore merits further investigation with a view to potential chemotherapeutic applications.

Nisin, a bacteriocin and commonly used food preservative, may serve as a novel potential therapeutic for treating head and neck squamous cell carcinoma (HNSCC), as it induces preferential apoptosis, cell cycle arrest, and reduces cell proliferation in HNSCC cells, compared with primary keratinocytes. Nisin also reduces HNSCC tumorigenesis in vivo. Mechanistically, nisin exerts these effects on HNSCC, in part, through CHAC1, a proapoptotic cation transport regulator, and through a concomitant CHAC1-independent influx of extracellular calcium. In addition, although CHAC1 is known as an apoptotic mediator, its effects on cancer cell apoptosis have not been examined. Our studies are the first to report CHAC1's new role in promoting cancer cell apoptosis under nisin treatment. These data support the concept that nisin decreases HNSCC tumorigenesis in vitro and in vivo by inducing increased cell apoptosis and decreased cell proliferation; effects that are mediated by activation of CHAC1, increased calcium influxes, and induction of cell cycle arrest. These findings support the use of nisin as a potentially novel therapeutic for HNSCC, and as nisin is safe for human consumption and currently used in food preservation, its translation into a clinical setting may be facilitated.

Antimicrobial delivery systems have been proposed as potential solutions to improve effectiveness of antimicrobials in food matrixes by shielding antimicrobials from contacting food matrix components and releasing them continuously. In this work, spray-dried capsules were produced from zein solutions with the same concentrations of nisin and thymol but with varying Tween 20 contents for characterization of release kinetics of antimicrobials and antilisterial properties. At intermediate levels of Tween 20, sustained and more complete release of antimicrobials was observed at pH 6.0 and 8.0. Most capsule samples were more effective than free antimicrobials against Listeria monocytogenes in 2% reduced fat milk, and the best capsule treatment reduced the bacterial population by 2 log CFU/mL more than comparable free antimicrobials after 4 h incubation at 25 °C. Our work demonstrated that nonionic surfactant can be conveniently used to modulate characteristics of delivery systems to effectively improve antimicrobial functions in food systems.

Aims: To determine if nisin F has an effect on the bacterial population in the gastro-intestinal tract., Methods and Results: Six male C57BL/6 mice were intraperitoneally injected with 200 μl sterile saline and six with nisin F (200 μl, equivalent to 640 arbitrary units). Fecal samples were collected before injection and 8, 24 and 48 h after injection, and the bacteria amplified by PCR-DGGE using 16S rDNA primers. The composition of the bacterial population in the gastro-intestinal tract (GIT) of mice that were injected with saline changed during 48 h, whereas the bacterial population in the GIT remained relatively unchanged in animals injected with nisin F., Conclusions: These results suggest that nisin F inhibits the growth of specific bacteria in the GIT within the first 4 h. Furthermore, the species remained repressed for at least 44 h after one intraperitoneal injection with nisin F., Significance and Impact of the Study: This is the first report suggesting that nisin F may have a stabilizing effect on the bacterial population in the gastro-intestinal tract., (© 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.)

Food grade antimicrobial delivery systems were studied in this work to enhance the effectiveness of antimicrobials inhibiting the growth of Listeria monocytogenes during storage. Corn zein was used as a carrier biopolymer and nisin and thymol as antimicrobials. Capsules produced by spray drying demonstrated different microstructures and release characteristics of nisin at different usage levels of thymol. Better release profiles were achieved when glycerol was additionally used to prepare capsules. Capsules showing sustained release of significant amounts of both antimicrobials effectively inhibited the growth of L. monocytogenes at pH 6.0 and 30 °C in the growth medium. Capsules were also more effective than free antimicrobials in inhibiting the growth of L. monocytogenes in 2% reduced fat milk at 25 °C. Our work showed that engineered delivery systems have promise to fulfill the antimicrobial effectiveness during shelf life storage of foods to ensure microbiological safety.

In this work, carbohydrate nanoparticles were created to prolong the efficacy of antimicrobial peptide against pathogens. Nisin and Listeria monocytogenes were used as the peptide and pathogen models, respectively, and phytoglycogen (PG)-based nanoparticles were developed as carriers of nisin. PG from su1 mutant maize was subjected to β-amylolysis as well as subsequent succinate or octenyl succinate substitutions. The goal was to minimize the loss of peptide during storage and meanwhile realize an effective release in the presence of bacteria. The capabilities of PG derivatives as carriers of nisin were evaluated using centrifugal ultrafiltration, zeta-potential, and the initial availability of nisin against L. monocytogenes. All methods indicated that nisin loading was favored by a high degree of substitution (DS), presence of hydrophobic octenyl moiety, and β-amylolysis of PG nanoparticles. To evaluate the prolonged nisin efficacy, preparations containing nisin and PG derivatives were loaded into a BHI-agar deep-well model (mimicking nisin depletion at the nutrient-containing surface). The residual inhibitory activities of preparations against L. monocytogenes were monitored during 21 days of storage at 4 °C. The results showed that all PG derivatives led to the prolonged retention of nisin activity and the longest retention was associated with high DS, β-amylolysis, and octenyl succinate. Evidently, both electrostatic and hydrophobic interactions are the driving forces of nisin adsorption, and the glucan structure at the nanoparticle surface also affects nisin loading and retention during storage., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Background: We investigated the in vitro activities of nisin alone or in combination with vancomycin and ciprofloxacin against methicillin-resistant (MRSA) and -susceptible Staphylococcus aureus (MSSA) strains., Methods: The minimum inhibitory concentrations were determined by microbroth dilution technique. Antibiotic combinations were assessed using the checkerboard technique. The time-kill curve method was used for determining the bactericidal activity of nisin alone and in combination., Results: For both MSSA and MRSA strains, the minimum inhibitory concentrations of nisin ranged between 4 and 16 mg/l. With a fractional inhibitory concentration of ≥0.5 as borderline, synergistic interactions were seen in three of five isolates with nisin-ciprofloxacin compared to two of five isolates with nisin-vancomycin combinations against both MSSA and MRSA. No antagonism was observed. The results of time-kill curve analysis demonstrated concentration-dependent rapid bactericidal activity of nisin and synergism almost in all strains when nisin was used in combination with ciprofloxacin, and early synergistic interactions in some of the strains when it was used in combination with vancomycin., Conclusion: Nisin seems to be a good candidate for further investigations in the treatment of Gram-positive bacteria, alone or in combination with antibiotics., (Copyright © 2012 S. Karger AG, Basel.)

The effect of different concentrations of Zataria multiflora Boiss. essential oil (EO; 0%, 0.005%, and 0.015%), nisin (0, 0.125, and 0.25 microL/mL), and their combinations on the production of staphylococcal enterotoxin C (SEC) and alpha-hemolysin (alpha-toxin) by Staphylococcus aureus at different inoculation levels (10(3), 10(4), and 10(5) cfu/mL) in brain heart infusion broth during storage at 35 degrees C for up to 43 days was evaluated. The SEC production was significantly (p < 0.05) inhibited and the hemolysis due to alpha-toxin was significantly (p < 0.05) reduced by EO concentration at levels 0.015% and 0.005%, respectively. Significant (p < 0.05) inhibitory effect of EO on SEC production at level 0.005% was observed when it was used in combination with nisin = 0.125 microL/mL. The significant (p < 0.05) synergistic effect of EO = 0.005% and nisin = 0.125 microL/mL was also observed as more reduction of hemolysis due to alpha-toxin than EO = 0.005% alone. Further, EO significantly (p < 0.05) prevented SEC production by S. aureus during the manufacturing process of a traditional Iranian white brined cheese (as a food model) even at its lowest concentration (5 microL/100 mL), in this study.

The effect of daily supplementation of nisin (2 mg/L), monensin (5.88 mg/L) and nisin and monensin (2 + 5.88 mg/L) on ovine ruminal ciliates and bacteria was investigated using the artificial rumen RUSITEC. Major groups in RUSITEC were Entodinium spp. and Dasytricha ruminantium. The supplementation of nisin significantly increased the population of both major ciliate groups. The supplementation of monensin significantly decreased the population of both groups. The combined effect of nisin and monensin was similar to the effect of monensin. Monensin had strong antiprotozoic effects in contrast to the stimulatory effects of nisin. D. ruminantium followed by Entodinium spp. appeared more resistant to tested compounds than other rumen ciliates. Tested additives did not significantly influence the presence and growth of amylolytic streptococci and enterococci but nisin showed a tendency to decreasing the concentration of Escherichia coli and lactobacilli.

Background: Nisin, a naturally occurring antimicrobial peptide (AMP), is currently the focus of clinical trials as an intravaginal microbicide. Therefore its mechanism of interaction with various cell membranes was studied., Study Design: Flow cytometry was used for quantitative estimation of membrane damage by nisin which was further determined by scanning electron microscopy (SEM). Affinity of nisin for different unilamellar liposome vesicles was determined spectroflurometrically and confirmed using laser scanning confocal microscopy (LSCM)., Results: Propidium iodide (PI) staining by flow cytometry exhibited selective membrane permeabilizing effect of nisin on sperm and bacterial membranes which correlated with ultrastructural changes. In vitro interaction of nisin with liposome model vesicles revealed significant leakage of calcein from liposomes composed of phosphatidylcholine/phosphatidylglycerol (POPC/POPG) (e.g., bacteria) and phosphatidylcholine/phosphatidylserine (POPC/POPS) (e.g., spermatozoa) as compared to phosphatidylcholine/phosphatidylethanolamine (POPC/POPE) vesicles (e.g., red blood corpuscles). LSCM results were in complete agreement with cell membrane affinity studies., Conclusion: This unique property of nisin can be exploited in the development of a safe and effective vaginal microbicide for the prevention of sexually transmitted infections/acquired immunodeficiency syndrome (STIs/AIDS) and unplanned pregnancies.

The in vivo evaluation of colon-targeting tablets was conducted in six healthy male volunteers. A pectin-hydroxypropyl methylcellulose coating was compressed onto core tablets labelled with 4MBq (99m)Tc-DTPA. The tablets released in the colon in all subjects; three in the ascending colon (AC) and three in the transverse colon (TC). Tablets that released in the TC had reached the AC before or just after food (Group A). The other three tablets released immediately upon AC entry at least 1.5h post-meal (Group B). Release onset for Group B was earlier than Group A (343min vs 448min). Group B tablets exhibited a clear residence period at the ileocaecal junction (ICJ) which was not observed in Group A. Prolonged residence at the ICJ is assumed to have increased hydration of the hydrogel layer surrounding the core tablet. Forces applied as the tablets progressed through the ICJ may have disrupted the hydrogel layer sufficiently to initiate radiolabel release. Conversely, Group A tablets moved rapidly through the AC to the TC, possibly minimising contact times with water pockets. Inadequate prior hydration of the hydrogel layer preventing access of pectinolytic enzymes and reduced fluid availability in the TC may have retarded tablet disintegration and radiolabel diffusion.

Aims: To determine the antimicrobial activity of nisin F against Staphylococcus aureus in the respiratory tract., Methods and Results: The respiratory tract of nonimmunosuppressed and immunosuppressed Wistar rats were colonized with 4 x 10(5) viable cells of S. aureus K and then treated by administering 8192 arbitrary units (AU) nisin F intranasal. Symptoms of pneumonia were detected in the trachea and lungs of immunosuppressed rats that had not been treated with nisin F. The trachea and lungs of immunosuppressed rats treated with nisin F were healthy. No significant differences were recorded in blood cell indices. The antimicrobial activity of low concentrations nisin F (80-320 AU ml(-1)) was slightly stimulated by lysozyme and lactoferrin., Conclusions: Nisin F inhibited the growth of S. aureus K in the respiratory tract of immunocompromised rats. Treatment with nisin F at 8192 AU proofed safe, as the trachea, lungs, bronchi and haematology of the rats appeared normal., Significance and Impact of the Study: Nisin F is nontoxic and may be used to control respiratory tract infections caused by S. aureus. This is, however, a preliminary study with an animal model and need to be confirmed with studies on humans.

The effectiveness of whey protein isolate (WPI) coatings incorporated with grape seed extract (GSE), nisin (N), malic acid (MA), and ethylenediamine tetraacetic acid (EDTA) and their combinations to inhibit the growth of Listeria monocytogenes, E. coli O157:H7, and Salmonella typhimurium were evaluated in a turkey frankfurter system through surface inoculation (approximately 10(6) CFU/g) of pathogens. The inoculated frankfurters were dipped into WPI film forming solutions both with and without the addition of antimicrobial agents (GSE, MA, or N and EDTA, or combinations). Samples were stored at 4 degrees C for 28 d. The L. monocytogenes population (5.5 log/g) decreased to 2.3 log/g after 28 d at 4 degrees C in the samples containing nisin (6000 IU/g) combined with GSE (0.5%) and MA (1.0%). The S. typhimurium population (6.0 log/g) was decreased to approximately 1 log cycles after 28 d at 4 degrees C in the samples coated with WPI containing a combination of N, MA, GSE, and EDTA. The E. coli O157:H7 population (6.15 log/g) was decreased by 4.6 log cycles after 28 d in samples containing WPI coating incorporated with N, MA, and EDTA. These findings demonstrated that the use of an edible film coating containing nisin, organic acids, and natural extracts is a promising means of controlling the growth and recontamination of L. monocytogenes, S. typhimurium, and E. coli O157:H7 in ready-to-eat poultry products.

Topical microbicides are intended for frequent use by women in reproductive age. Hence, it is essential to evaluate their impact on mucosal immune function in the vagina. In the present study, we evaluated nisin, a naturally occurring antimicrobial peptide (AMP), for its efficacy as an intravaginal microbicide. Its effect on the vaginal immune function was determined by localizing Toll-like receptors (TLRs-3, 9) and cytokines (IL-4, 6 , 10 and TNF-alpha) in the rabbit cervicovaginal epithelium following intravaginal administration of high dose of nisin gel for 14 consecutive days. The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels. However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade. Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides.

Nisin containing pectin/HPMC compression coated tablets were prepared and their in vitro behavior tested for colonic delivery. Nisin is a 34-amino-acid residue long, heat stable peptide belonging to the group A lantibiotics with wide antimicrobial activity against Gram-positive bacteria. The invention can be useful for treating colonic infectious diseases such as by Clostridium difficile, and also by colonization of vancomycin-resistant enterococci. In this study, each 100mg core tablet of nisin was compression coated with 100% pectin, 90% pectin-10% HPMC, 85% pectin-15% HPMC, 80% pectin-20% HPMC, 75% pectin-25% HPMC, 100% HPMC at a coat weight of 400mg. The concentration and the activity of nisin were quantified using Well Diffusion Agar Assay. Drug release studies were carried out in pH 3.3 buffer solution. System degradation/erosion experiments were carried out in pH 1.2, 3.3, and 6.8 buffers using a pectinolytic enzyme. The biological activity and NMR studies were performed to assess the stability of nisin during the processing and after the in vitro tests. It was found that pectin alone was not sufficient to protect the nisin containing core tablets. At the end of the 6h 40% degradation was observed for 100% pectin tablets. HPMC addition required to control the solubility of pectin, a 5% increase in HPMC ratio in pectin/HPMC mixture provided a 2-h lag time for nisin release. Eighty percent pectin-20% HPMC appeared to be an optimum combination for further evaluation. Tablets maintained their integrity during the 6-h dissolution test, approximating the colon arrival times. Nisin was found to be active/stable during processing and after in vitro tests. Effect of polymer hydration on pectin degradation was found to be crucial for the enzyme activity. Sufficiently hydrated pectin degraded faster. The pectin/HPMC envelope was found to be a good delivery system for nisin to be delivered to the colon.

Nisin is an antimicrobial polypeptide produced by Lactococcus lactis and is believed nontoxic to humans. The objective of this study was to evaluate a nisin-based formulation for the treatment of bovine clinical mastitis in lactating dairy cattle. A total of 92 cows with 107 clinically mastitic quarters were randomly assigned to nisin- (48 cows with 51 quarters) and gentamicin (GM)-treated (44 cows with 56 quarters) groups. In the nisin-treated group, cows received an intramammary infusion of nisin at a dose of 2,500,000 IU; in the GM-treated group, intramammary infusion of GM was administered at a dose of 0.8 g. Results indicated that nisin offered a clinical cure rate similar to GM (90.2 vs. 91.1%) and no difference in bacteriological cure rate than GM-treated group (60.8 vs. 44.6%, respectively). Proportion of the quarters with milk somatic cell counts <500,000 cells/mL was not different in the nisin-treated group (50.0 and 47.8%) compared with the GM-treated group (33.3 and 37.3%) 1 and 2 wk after treatment. Of 17 Staphylococcus aureus isolates, 82.5% were resistant to penicillin, and 35.3% to GM, but none of them to nisin. Nisin therapy eliminated 54.5% (6 of 11) of S. aureus IMI, whereas GM eliminated 33.3% (2 of 6). Nisin in milk (4.5 +/- 0.8 IU/mL) was detected only at 12 h following intramammary infusion, which was much lower than the upper limit (500 mg/mL) allowed as preservative in milk by the China authority. Because of its efficacy in the treatment of bovine clinical mastitis, especially resistant Staph. aureus-caused IMI, as well as its safety in humans, nisin deserves further study to clarify its effects on mastitis caused by different mastitis pathogens on a larger scale.

Aims: The fate of Listeria monocytogenes Scott A, was studied in broth, at different a(w)s (by adding NaCl or KCl from 0.0 to 1.4 mol l(-1)), pHs (from 4.0 to 7.3 by adding lactic acid), and nisin concentrations (from 0 to 100 IU ml(-1))., Methods and Results: Increasing salt and nisin concentrations and decreasing pH resulted in lower growth rates and extended lag phases. At pH 4.5 no growth was observed while in presence of nisin and/or 1 mol l(-1) salts of both kinds, L. monocytogenes Scott A was inactivated. Equal-molar concentrations of NaCl or KCl (similar a(w)), exerted similar effects against L. monocytogenes in terms of lag phase duration, growth or death rate. The growth boundaries of L. monocytogenes Scott A at 5 degrees C were also estimated by growth/no growth turbidity data, modeled by logistic polynomial regression. The concordance of logistic models, were 99.6 and 99.8% for NaCl and KCl, respectively., Conclusions: The growth interfaces derived by both NaCl and KCl models were almost identical. Hence, NaCl can be replaced by KCl without risking the microbiological safety of the product. Increasing nisin concentrations markedly affected the interface resulting in a more inhibitory environment for L. monocytogenes Scott A. Low to medium salt concentrations (0.3-0.7 mol l(-1) of either NaCl or KCl) provided a protective effect against inhibition of L. monocytogenes Scott A by nisin., Significance and Impact of the Study: Modelling the growth boundaries not only contributes to the development of safer food by providing useful data, but can also be used to study interactions between factors affecting initiation of growth of pathogenic micro-organisms.

Nanoencapsulation may improve activity of protein or polypeptide antimicrobials against a variety of microorganisms. In this study, nanoliposomes prepared from different lipids (Phospholipon 90H, Phospholipon 100H, dipalmitoylphosphatidylcholine (DPPC), stearylamine (SA), dicetyl phosphate (DCP) and cholesterol) by a new, non-toxic and scalable method, were tested for their capacity to encapsulate nisin Z and target bacteria (Bacillus subtilis and Pseudomonasaeruginosa). Factors affecting the entrapment efficiency (charge and cholesterol concentration in the vesicles) and stability of nanoliposomes were assessed. The nanoliposomes and their bacterial targeting were visualised, using different microscopes under air and liquid environments. Nisin was entrapped in different nanoliposomes with encapsulation efficiencies (EE) ranging from 12% to 54%. Anionic vesicles possessed the highest EE for nisin while increase in cholesterol content in lipid membranes up to 20% molar ratio resulted in a reduction in EE. Stability of nanoliposome-encapsulated nisin was demonstrated for at least 14 months at 4 degrees C (DPPC:DCP:CHOL vesicles) and for 12 months at 25 degrees C (DPPC:SA:CHOL vesicles).