Alicyclobacillus spp. is a potential spoiling agent of acidic products and citrus drinks, leading to sensory alterations in contaminated products and consequent economic losses. Treatments such as pasteurization eliminate vegetative cells, but also create a favorable atmosphere for spore germination. To guarantee quality and safety, the application of natural substances as bioconservatives is a considerable and promising alternative for the food industry. This study evaluated the effect of hexane extract of Matricaria chamomilla L. (HE), Nisin (N) and their combination (HE + N). These compounds are present in some studies describing their antibacterial action, but no studies were found on the association of these compounds against the species Alicyclobacillus spp. This study aimed to analyze the antioxidant activity (AA) for the DPPH• (0,23 μmol Trolox/mg) and ABTS (27.93 μmol Trolox/mg), the Checkboard test revealed synergism between HE and N with a fractional inhibitory index (FIC) of 0.068., and to study the antibacterial and sporicidal effect. The antibacterial and sporicidal activity was satisfactory against Alicyclobacillus acidoterrestris with MIC and MBC of 1.95 μg/mL and MSC of 7.81 μg/mL in analyzes using HE + N. The application in orange juice proved to be effective, with an MBC of 0.007 μg/mL. The MIC results served as a parameter for other tests carried out in this study, such as flow cytometry and Scanning Electron Microscopy (SEM), and for the evaluation of sensory characteristics with Electronic Nose (E-nose).

Aim To evaluate and compare the antimicrobial efficacy of nisin in different carriers against Enterococcus faecalis. Materials and methods Test materials were divided into four groups of five samples each as follows: group 1 = nisin + 17% ethylenediaminetetraacetic acid (EDTA); group 2 = nisin + 2% chitosan; group 3 = nisin + 2% chlorhexidine; group 4 = nisin + distilled water (control). The antimicrobial effectiveness was assessed using the direct contact method, where a standardized E. faecalis suspension was applied to the test materials. Optical density (OD) was assessed using enzyme-linked immunosorbent assay (ELISA) at the end of days one and seven. Data were analyzed using ANOVA and Tukey\'s post hoc analysis. The level of significance was set at p < 0.05. Results On day one, there was a significant difference in the mean OD values (p < 0.001) with group 3 showing the highest, followed by groups 1, 2, and 4. On day seven, all groups demonstrated antibacterial activity (group 1 > group 3 > group 4 > group 2) but the differences were not statistically significant (p = 0.393). Intragroup analysis showed a decrease in the OD values from day one to day seven, the difference of which was not significant in all groups except group 1, which showed a significant difference (p = 0.035). Conclusion The antibacterial efficacy of nisin was synergistically enhanced with the addition of 17% EDTA and 2% chlorhexidine over seven days against E. faecalis.

The accumulation of nisin in the fermentation medium can reduce the process\'s productivity. This research studied the potential of Nymphaea alba leaf powder (NALP) as a hydrophobic biosorbent for efficient in-situ nisin adsorption from the fermentation medium by docking and experimental analysis. Molecular docking analysis showed that di-galloyl ellagic acid, a phytochemical compound found in N. alba, had the highest affinity towards nisin. Enhancements in nisin adsorption were seen following pre-treatment of NAPL with HCl and MgCl2. A logistic growth model was employed to evaluate the growth dynamics of the biosorption capacity, offering valuable insights for process scalability. Furthermore, optimization through Response Surface Methodology elucidated optimal nisin desorption conditions by Liebig\'s law of the minimum, which posits that the scarcest resource governs production efficiency. Fourier Transform Infrared (FTIR) spectroscopy pinpointed vital functional groups involved in biosorption. Scanning electron microscopy revealed the changing physical characteristics of the biosorbent after exposure to nisin. The findings designate NALP as a feasible adsorbent for nisin removal from the fermentation broth, thus facilitating its application in the purification of other biotechnological products based on growth and production optimization principles.

Periodontitis-related oral microbial dysbiosis is thought to contribute to adverse pregnancy outcomes (APOs), infertility, and female reproductive inflammation. Since probiotics can modulate periodontitis and oral microbiome dysbiosis, this study examined the effects of a probiotic bacteriocin, nisin, in modulating the reproductive microbiome and inflammation triggered by periodontitis. A total of 24 eight-week-old BALB/cByJ female mice were randomly divided into four treatment groups (control, infection, nisin, and infection+nisin group), with 6 mice per group. A polymicrobial (Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum) mouse model of periodontal disease was used to evaluate the effects of this disease on the female reproductive system, with a focus on the microbiome, local inflammation, and nisin\'s therapeutic potential in this context. Moreover, 16s RNA sequencing was used to evaluate the changes in the microbiome and RT-PCR was used to evaluate the changes in inflammatory cytokines. Periodontal pathogen DNA was detected in the reproductive organs, and in the heart and aorta at the end of the experimental period, and the DNA was especially elevated in the oral cavity in the infection group. Compared to the control groups, only P. gingivalis was significantly higher in the oral cavity and uterus of the infection groups, and T. forsythia and F. nucleatum were significantly higher in the oral cavity of the infection groups. The infection and nisin treatment group had significantly lower levels of P. gingivalis, T. forsythia, and F. nucleatum in the oral cavity compared with the infection group. Since periodontal pathogen DNA was also detected in the heart and aorta, this suggests potential circulatory system transmission. The polymicrobial infection generally decreased the microbiome diversity in the uterus, which was abrogated by nisin treatment. The polymicrobial infection groups, compared to the control groups, generally had lower Firmicutes and higher Bacteroidota in all the reproductive organs, with similar trends revealed in the heart. However, the nisin treatment group and the infection and nisin group, compared to the control or infection groups, generally had higher Proteobacteria and lower Firmicutes and Bacteroidota in the reproductive organs and the heart. Nisin treatment also altered the microbiome community structure in the reproductive tract to a new state that did not mirror the controls. Periodontal disease, compared to the controls, triggered an increase in inflammatory cytokines (IL-6, TNF-α) in the uterus and oral cavity, which was abrogated by nisin treatment. Polymicrobial periodontal disease alters the reproductive tract\'s microbial profile, microbiome, and inflammatory status. Nisin modulates the microbial profile and microbiome of the reproductive tract and mitigates the elevated uterine inflammatory cytokines triggered by periodontal disease.

The wild-type Lactococcus lactis strain LAC460 produces two bacteriocin-like phage lysins, LysL and LysP. This study aimed to produce and secrete LysL in various heterologous hosts and an in vitro cell-free expression system for further functional studies. Initially, the lysL gene from L. lactis LAC460 was cloned into Lactococcus cremoris NZ9000 and L. lactis N8 strains, with and without the usp45 signal sequence (SSusp45), under a nisin-inducible promoter. Active LysL was primarily produced intracellularly in recombinant L. lactis N8, with some secretion into the supernatant. Recombinant L. cremoris NZ9000 lysed upon nisin induction, indicating successful lysL expression. However, fusion with Usp45 signal peptide (SPUsp45-LysL) weakened LysL activity, likely due to incomplete signal peptide cleavage during secretion. Active LysL was also produced in vitro, and analysed in SDS-PAGE, giving a 42-kDa band. However, the yield of LysL protein was still low when produced from recombinant lactococci or by in vitro expression system. Therefore, His-tagged LysL was produced in Escherichia coli BL21(DE3). Western blot confirmed the intracellular production of about 44-kDa His-tagged LysL in E. coli. His-tagged active LysL was then purified by Ni-NTA affinity chromatography yielding sufficient 4.34 mg of protein to be used in future functional studies.

UNASSIGNED: This study was conducted to compare the effects of nisin (NIS) and ionophore antibiotic monensin (MON) on the growth performance, rumen fermentation, nutrient digestion and plasma metabolites of fattening Hu sheep.
UNASSIGNED: Thirty-six male Hu sheep (23.5 ± 1.0 kg) were divided into two blocks based on BW (low BW and high BW). Sheep within each block were then allotted to 9 pens respectively (two sheep/pen). Pens within each block were randomly assigned to one of three dietary treatments: (1) basal diet (CON); (2) basal diet + 40 mg/kg DM of MON; (3) basal diet + 274.5 mg/kg DM of NIS. The study lasted 9 weeks, with the initial 2 weeks for adaptation and the subsequent 7 weeks for treatment.
UNASSIGNED: The results showed that both NIS and MON addition had no impacts on average daily gain (ADG), dry matter intake (DMI), and feed conservation rate (G:F) of sheep (p > 0.05). The digestibility of ether extract (EE) was lower in the MON-fed and NIS-fed sheep (p < 0.01) than in the CON group, whereas crude protein (CP) digestibility was higher in the MON-fed sheep compared to those fed NIS (p < 0.05). Both NIS and MON supplementation decreased acetate levels and acetate/propionate ratio in the rumen of Hu sheep (p < 0.05). Sheep fed MON exhibited higher total cholesterol concentrations (p < 0.05) compared to the CON and NIS groups. However, there were no significant differences in other plasma metabolites, including blood urea nitrogen (BUN), total bile acid, triglyceride, total protein, albumin, globulin, glucose, etc., among the three groups (p > 0.05).
UNASSIGNED: In conclusion, dietary addition of NIS and MON altered the rumen fermentation mode by reducing acetate levels, with no discernible effects on the growth performance of the fattening Hu sheep.

In this study, Listeria monocytogenes from minced pork was evaluated for changes in resistance to thermal treatment and gastric fluid following environmental stresses during food processing. Bacteria were exposed to cold stress, followed by successive exposures to different stressors (lactic acid (LA), NaCl, or Nisin), followed by thermal treatments, and finally, their gastrointestinal tolerance was determined. Adaptation to NaCl stress reduced the tolerance of L. monocytogenes to subsequent LA and Nisin stress. Adaptation to LA stress increased bacterial survival in NaCl and Nisin-stressed environments. Bacteria adapted to Nisin stress showed no change in tolerance to subsequent stress conditions. In addition, treatment with NaCl and LA enhanced the thermal tolerance of L. monocytogenes, but treatment with Nisin decreased the thermal tolerance of the bacteria. Almost all of the sequential stresses reduced the effect of a single stress on bacterial thermal tolerance. The addition of LA and Nisin as a second step of stress reduced the tolerance of L. monocytogenes to gastric fluid, whereas the addition of NaCl enhanced its tolerance. The results of this study are expected to inform processing conditions and sequences for meat preservation and processing and reduce uncertainty in risk assessment of foodborne pathogens due to stress adaptation.

The exceptional biodegradability and active biological functions of bio-based packaging materials have attracted increasing interest. In this study, a bioplastic film was developed by introducing simultaneously polyphenols (tea polyphenols, TPs) and peptides (nisin) into a soy protein isolate/sodium alginate (SPI/SA) based film-forming matrix. The research results revealed that the dynamic coordinated interaction between TPs and nisin enhanced mechanical properties, UV-resistance, and thermal stability of bioplastic films. Furthermore, the bioplastic film exhibited antibacterial activity and antioxidant properties. Significantly, biofilm growth of Staphylococcus aureus treated with TPs-5/Nisin-5 bioplastic film was inhibited by 91.12% compared to the blank group. The shelf life of beef with TPs-5/Nisin-5 bioplastic film was prolonged by 2 days because of the synergistic effect of TPs and nisin. Additionally, the bioplastic film biodegraded in the natural environment about 21 days. This environmentally friendly regeneration strategy and the integration of advantageous functions provided ideas for the development of active food packaging.

BACKGROUND: Antibiotic resistance is a global threat to human health that leads to disasters. Acinetobacter baumannii cannot be controlled by the existing antibiotics, and it became challenging. Therefore, novel antibacterial agents are required to combat such threats. The aim of this project is to find a novel antimicrobial agent to treat this multi-drug resistant bacterium.
METHODS: The NisA gene was isolated from Lactococcus lactis spp. lactis and cloned into the pET-3a plasmid using Gibson cloning assembly. Purified Nisin from cloning was conjugated with silver nanoparticles. Finally, an assessment of antibacterial activity for each of the purified Nisin, Silver nanoparticles, and Nisin-Silver nanoparticles conjugate against the extensively drug-resistant A. baumannii was performed.
RESULTS: Nisin was successfully purified from cloned bacteria, and the concentration was 416 µg/ml. The conjugation of nisin and silver nanoparticles was analyzed by electron microscopy. The minimum inhibitory concentration of Nisin and silver nanoparticles against A. baumannii were 104 µg/ml and 125 µg/ml, respectively. While Nisin-silver nanoparticle conjugates showed potent antimicrobial activity with MIC 125-52 µg/ml in which silver nanoparticles increased the antimicrobial activity of nisin beyond its optimum concentration (104 µg/ml). ‎ CONCLUSION: The development of new antibacterial agents is necessary to control extensively drug-resistant bacteria. Nisin-silver conjugates showed more potent antimicrobial activity than when applied separately and gave hope to combat the multi-drug resistant A. baumannii.

Edible films are effective alternatives to plastic packaging, however, the hydrophilicity of edible films based on protein and polysaccharide limits the application. Therefore, we fabricated a water-stable hybrid film with a linear-spherical interpenetrating molecular topology network using egg white (EW), chitosan (CS), and pectin. Meanwhile, the nisin-tannin acid self-assembly complex nanoparticles were employed as a multifunctional cross-linker, antibacterial and antioxidant agent to improve the performance of films. The FTIR, XRD, and SEM analysis revealed that the conformation and crystalline structure rearrangement of chitosan induced by the alkaline environment provided by egg white enhanced the network structure of films, effectively avoided the addition of modifying reagents. The proposed hybrid films exhibited excellent properties, with EW/TNPCS3 showing the best overall performance. The water contact angle (WCA) increased to 105.27 ± 1.62°, and its dissolution and swelling rates were significantly lower than pure egg white and pure chitosan films. Moreover, tannin-nisin (TN) nanoparticles endowed the films with excellent antimicrobial activity against the common Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. Thus, the prepared blending films have great application potential in food preservation, especially to maintain stable performance in high humidity environment.