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This study describes the discovery and characterization of raffinocyclicin, a novel plasmid-encoded circular bacteriocin, produced by the raw milk isolate Lactococcus raffinolactis APC 3967. This bacteriocin has a molecular mass of 6,092 Da and contains 61 amino acids with a three-amino acid leader peptide. It shows the highest identity to the circular bacteriocins bacicyclicin XIN-1 (42.62%), aureocyclicin 4185 (42.62%), and garvicin ML (41.53%). A broad inhibitory spectrum includes strains from Staphylococcus, Enterococcus, Streptococcus, Micrococcus, Lactobacillus, Leuconostoc, and Listeria, in addition to a pronounced inhibitory effect against Lactococcus and Clostridium. It displays low sensitivity to trypsin, most likely as a result of its circular nature. The raffinocyclicin gene cluster is composed of 10 genes: 6 core genes, genes encoding an accessory three-component ABC transporter (rafCDE), and a putative transcriptional regulator related to the MutR family. A lack of inhibitory activity in the cell-free supernatant combined with the pronounced activity of cell extracts suggests that the majority of raffinocyclicin is associated with the cell rather than being released to the extracellular environment. This is the first report of a bacteriocin produced by the L. raffinolactis species.IMPORTANCEThe present study aimed to characterize raffinocyclicin, a novel circular bacteriocin produced by the lactic acid bacteria Lactococcus raffinolactis APC 3967. Bacteriocins are generally cationic and hydrophobic peptides with antimicrobial activity, which present diverse biotechnological properties of interest for the food industry. Raffinocyclicin inhibits a wide range of bacteria, including foodborne pathogens, and is stable against different treatments which suggest its potential as a natural biopreservative. Whole-genome sequencing and the genetic analysis of the raffinocyclicin gene cluster showed that it is encoded by plasmid that could be used in the future to transfer the ability to produce the bacteriocin to other lactic acid bacteria for industrial applications. These results together highlight the potential of this novel antimicrobial as a biopreservative to be used by the food industry.
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Global food security has recently been under serious threat from the rapid rise in the world's population, the problems brought on by climate change, and the appearance of new pandemics. As a result, the need for novel and innovative solutions to solve the existing problems and improve food sustainability has become crucial. 3D printing is expected to play a significant role in providing tangible contributions to the food industry in achieving sustainable development goals. The 3D food printing holds the potential to produce highly customized food in terms of shape, texture, flavor, structure and nutritional value and enable us to create new unique formulations and edible alternatives. The problem of whether the cost of the printed meal and 3D printing itself can be sustainably produced is becoming more and more important due to global concerns. This review intends to provide a comprehensive overview of 3D printed foods with an overview of the current printing methodologies, illustrating the technology's influencing factors, and its applications in personalized nutrition, packaging, value addition, and valorization aspects to fully integrate sustainability concerns thus exploring the potential of 3D food printing.
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The storage mite Tyrophagus putrescentiae infests a wide range of food products including pet food. Control of this mite depends on chemical methods such as fumigation and spraying with insecticides. Methyl bromide was used as a fumigant for high-value stored products, especially to control mite infestation in dry-cured hams and cheeses, but it is now banned for most uses in many countries because of its atmospheric ozone-depleting effects. Effective alternatives to methyl bromide are needed to develop integrated pest management programs for this pest. Liquid smoke is a naturally derived flavoring and preservative with known antimicrobial properties. The objective of this study was to investigate the efficacy of liquid smoke preparations, with varying phenol and carbonyl concentrations and pH, on the survivability and orientation behavior of T. putrescentiae in a model semi-moist pet food. The mite survivability assays using liquid smoke-treated and untreated semi-moist pet food samples indicated that there was no difference among treatments (p > 0.05) for mite infestation and survival. Two-choice behavioral assays using semi-moist pet food cubes dipped in varying concentrations (0%, 0.3%, 1%, 5%, 10%, 25%, 50%, or 100% v/v) of liquid smoke preparations found that some of the liquid smoke preparations containing medium to high carbonyl content repelled the mites. In conclusion, liquid smoke did not kill or inhibit the mite population growth in semi-moist pet food. However, some liquid smoke fractions containing medium to high carbonyl content were repellent to mites and may retard mite infestation in stored semi-moist foods.
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Liquid smoke is a naturally derived flavor component and preservative with known antimicrobial properties. To our knowledge, there is a paucity of information on antifungal potential of liquid smoke against toxigenic fungi like Aspergillus flavus that produce mycotoxins in human and pet foods. Semi-moist pet food with high moisture content (20-30%) is susceptible to mold contamination and requires intervention. The objectives of this study were to determine the effects of liquid smoke preparations on the growth of wild-type mold and A. flavus in semi-moist pet food. Semi-moist pet food was formulated with eight different liquid smoke preparations (S1-S8) containing varying amounts of organic acids, phenol and carbonyl compounds (ranging from low to high) at 0% (untreated), 0.5, 1, 2, and 4% (w/w). A positive control consisted of 0.2% potassium sorbate known to inhibit mold growth. Shelf life was estimated by storing the samples at 28°C and 65-70% RH over 30 days and recording the number of days until the appearance of visible wild-type mold. In another experiment, samples were spot inoculated with A. flavus (â¼10,000 CFU/mL), incubated at 25°C, and analyzed for fungal growth at sampling intervals of 2 days over a 35-day period. Liquid smoke at 0.5, 1, 2, and 4% extended the shelf life of samples on an average by a total of 11.6, 12.5, 17.2, and 24.1 days when compared to the untreated samples (7.7 days). The smoke preparations Cloud S-C100 (S3) and Code-10 (S6) (high carbonyl, medium/low phenol) were the most effective (P < 0.05) in prolonging the number of days to visible mold growth (26-28 days). In the challenge study with A. flavus, Cloud S-C100 (S3), Cloud S-AC15 (S8) (high to medium carbonyl, low phenol), and Code 10 (S6) (base smoke) reduced (P < 0.05) mold counts by 1.0, 1.7, and 2.5 logs when compared to the untreated samples at 1, 2, and 4%, respectively. Addition of smoke at 0.5% did not reduce mold counts. The carbonyl preparations of liquid smoke were the most effective at enhancing shelf life of semi-moist pet food, and at inhibiting A. flavus growth.
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Post-processing operations of extruded pet food kibbles involve coating the product with fats and flavorings. These processes increase the risk for cross-contamination with food-borne pathogens such as Salmonella and Shiga toxin-producing Escherichia coli (STEC), and mycotoxin-producing molds such as Aspergillus spp. after the thermal kill step. In this study, the antimicrobial effects of two types of organic acid mixtures containing 2-hydroxy-4-(methylthio) butanoic acid (HMTBa), Activate DA™ and Activate US WD-MAX™, against Salmonella enterica, STEC and Aspergillus flavus when used as a coating on pet food kibbles were evaluated. Using canola oil and dry dog digest as fat and flavor coatings, the efficacy of Activate DA (HMTBa + fumaric acid + benzoic acid) at 0%, 1% and 2%, and Activate US WD-MAX (HMTBa + lactic acid + phosphoric acid) at 0%, 0.5% and 1% was tested on kibbles inoculated with a cocktail of S. enterica serovars (Enteritidis, Heidelberg and Typhimurium) or Shiga toxin-producing E. coli (STEC) serovars (O121, and O26) at 37 °C for 0, 12, 24, 48, 72 h, 30 and 60 days. Similarly, their efficacy was tested against A. flavus at 25 °C for 0, 3, 7, 14, 21, 28 and 35 days. Activate DA at 2% and Activate US WD-MAX at 1% reduced Salmonella counts by ~3 logs after 12 h and 4-4.6 logs after 24 h. Similarly, STEC counts were reduced by ~2 logs and 3 logs after 12 h and 24 h, respectively. Levels of A. flavus did not vary up to 7 days, and afterwards started to decline by >2 logs in 14 days, and up to 3.8-log reduction in 28 days for Activate DA and Activate US WD-MAX at 2% and 1%, respectively. The results suggest that the use of these organic acid mixtures containing HMTBa during kibble coating may mitigate post-processing enteric pathogen and mold contamination in pet food kibbles, with Activate US WD-MAX being effective at a lower concentration (0.5-1%) compared to Activate DA.
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Contaminated surfaces can transmit pathogens to food in industrial and domestic food-handling environments. Exposure to pathogens on food contact surfaces may take place via the cross-contamination of pathogens during postprocessing activities. Formaldehyde-based commercial sanitizers in recent years are less commonly being used within food manufacturing facilities due to consumer perception and labeling concerns. There is interest in investigating clean-label, food-safe components for use on food contact surfaces to mitigate contamination from pathogenic bacteria, including Salmonella. In this study, the antimicrobial effects of two types of organic acid mixtures containing 2-hydroxy-4-(methylthio) butanoic acid (HMTBa), Activate DA™ and Activate US WD-MAX™, against Salmonella when applied onto various food contact surfaces were evaluated. The efficacy of Activate DA (HMTBa + fumaric acid + benzoic acid) at 1% and 2% and Activate US WD-MAX (HMTBa + lactic acid + phosphoric acid) at 0.5% and 1% against Salmonella enterica (serovars Enteritidis, Heidelberg, and Typhimurium) were evaluated on six different material surfaces: plastic (bucket elevator and tote bag), rubber (bucket elevator belt and automobile tire), stainless steel, and concrete. There was a significant difference in the Salmonella log reduction on the material surfaces due to the organic acid treatments when compared to the untreated surfaces. The type of material surface also had an effect on the log reductions obtained. Stainless steel and plastic (tote) had the highest Salmonella log reductions (3-3.5 logs), while plastic (bucket elevator) and rubber (tire) had the lowest log reductions (1-1.7 logs) after treatment with Activate US WD-MAX. For Activate DA, the lowest log reductions (~1.6 logs) were observed for plastic (bucket elevator) and rubber (tire), and the highest reductions were observed for plastic (tote), stainless steel, and concrete (2.8-3.2 logs). Overall, the results suggested that Activate DA at 2% and Activate US WD-MAX at 1% are potentially effective at reducing Salmonella counts on food contact surfaces by 1.6-3.5 logs.
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Postbiotics are non-viable bacterial products or metabolic byproducts produced by probiotic microorganisms that have biologic activity in the host. Postbiotics are functional bioactive compounds, generated in a matrix during anaerobic fermentation of organic nutrients like prebiotics, for the generation of energy in the form of adenosine triphosphate. The byproducts of this metabolic sequence are called postbiotics, these are low molecular weight soluble compounds either secreted by live microflora or released after microbial cell lysis. A few examples of widely studied postbiotics are short-chain fatty acids, microbial cell fragments, extracellular polysaccharides, cell lysates, teichoic acid, vitamins, etc. Presently, prebiotics and probiotics are the products on the market; however, postbiotics are also gaining a great deal of attention. The numerous health advantages of postbiotic components may soon lead to an increase in consumer demand for postbiotic supplements. The most recent research aspects of postbiotics in the food and pharmaceutical industries are included in this review. The review encompasses a brief introduction, classification, production technologies, characterization, biological activities, and potential applications of postbiotics.
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The occurrence of recalls involving pathogenic Escherichia coli-contaminated wheat flours show the need for incorporating antimicrobial interventions in wheat milling. The objectives of this study were to assess the efficacy of sodium bisulfate (SBS) tempering in reducing E. coli O121 (ATCC 2219) and O26 (ATCC 2196) wheat load and to evaluate the impact of effective (≥3.0 log reductions) SBS treatments on wheat flour quality. Wheat grains were inoculated with E. coli (~6 log CFU/g) and tempered (17% moisture, 24 h) using the following SBS concentrations (%wheat basis): 0, 0.5, 0.75, 1.0, 1.25, and 1.5% SBS. Reductions in E. coli O121 and O26 wheat load at different time intervals (0.5, 2, 6, 12, 18, and 24 h) during tempering were evaluated. The addition of SBS during tempering resulted in E. coli (O121 and O26) log reductions of 2.0 (0.5% SBS) to >4.0 logs (1.5% SBS) (p ≤ 0.05). SBS tempering (1.25 and 1.5% SBS) produced acidic wheat flours (pH = 4.51-4.60) but had comparable wheat flour properties in terms of composition, dough, and bread-making properties relative to the control (0% SBS). SBS tempering reduced the E. coli O121 and O26 load of wheat after tempering with minimal effects on wheat flour quality.
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In recent years, several pet food recalls have been attributed to Salmonella contamination. In addition to the negative impacts on animal health, Salmonella-contaminated pet foods have been linked to infection in humans. With that in mind, the U.S. Food and Drug Administration has set forth a zero-tolerance policy for Salmonella in pet foods. Typically, pet foods are extruded or processed at high temperatures that are sufficient to reduce pathogenic bacteria. However, the possibility for postextrusion contamination still exists. One potential method to reduce the risk of postextrusion contamination of pet foods with Salmonella is through the addition of a chemical additive coating. The objective of this research was to evaluate the ability of ß-hydroxy-ß-methylbutyrate (HMB), in either free acid (HMBFA) or calcium salt (CaHMB) form, to reduce postextrusion contamination of dry extruded dog kibble with Salmonella. Three trials were conducted with HMBFA and CaHMB coated onto the kibbles at levels of 0, 0.1, 0.3, 0.5, 0.9, and 1.5% (w/w). The coated kibbles were then inoculated with Salmonella enterica subsp. enterica Enteritidis (ATCC 13076), with enumeration done on days 0, 1, 2, 7, and 14 postinoculation. Subsamples on each day were serially diluted, spread plated to xylose lysine deoxycholate agar, and incubated at 37°C for 24 h. Salmonella colonies were then counted and log CFU per gram was calculated. The 1.5% HMBFA reduced counts by 4.9 ± 0.2 log units on day 1, whereas the positive control only decreased 2.2 ± 0.1 log units (P < 0.0001). The 1.5% CaHMB level decreased counts by 7.1 ± 0.04 log units by day 7 compared with the control decrease of 2.1 ± 0.1 log units (P < 0.0001). All HMBFA and CaHMB treatments resulted in the elimination of detectable Salmonella counts by day 14 (P < 0.0001 versus controls). In conclusion, HMB coating was effective at reducing Salmonella artificially inoculated to dog kibbles in a model of postextrusion contamination.
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Manipulación de Alimentos/métodos , Salmonella enteritidis/efectos de los fármacos , Valeratos/farmacología , Animales , Recuento de Colonia Microbiana , Perros , Contaminación de Alimentos/prevención & control , Microbiología de Alimentos , Hemiterpenos , Humanos , Ácidos Pentanoicos , Salmonella enteritidis/crecimiento & desarrolloRESUMEN
Animal feed and ingredients are potential vectors of pathogenic bacteria. Contaminated ingredients can contaminate facility equipment, leading to cross-contamination of other products. This experiment was conducted to evaluate a standardized protocol for decontamination of an animal feed manufacturing facility using Enterococcus faecium (ATCC 31282) as an indicator. A pelleted swine diet inoculated with E. faecium was manufactured, and environmental samples (swabs, replicate organism detection and counting plates, and air samples) were collected (i) before inoculation (baseline data), (ii) after production of inoculated feed, (iii) after physical removal of organic material using pressurized air, (iv) after application of a chemical sanitizer containing a quaternary ammonium-glutaraldehyde blend, (v) after application of a chemical sanitizer containing sodium hypochlorite, (vi) after facility heat-up to 60 8 C for 24 h, (vii) for 48 h, and (viii) for 72 h. Air samples collected outside the facility confirmed pathogen containment; E. faecium levels were equal to or lower than baseline levels at each sample location. The decontamination step and its associated interactions were the only variables that affected E. faecium incidence (P < 0.0001 versus P > 0.22). After production of the inoculated diet, 85.7% of environmental samples were positive for E. faecium. Physical cleaning of equipment had no effect on contamination (P = 0.32). Chemical cleaning with a quaternary ammonium-glutaraldehyde blend and sodium hypochlorite each significantly reduced E. faecium contamination (P < 0.0001) to 28.6 and 2.4% of tested surfaces, respectively. All samples were negative for E. faecium after 48 h of heating. Both wet chemical cleaning and facility heating but not physical cleaning resulted in substantial E. faecium decontamination. These results confirmed both successful containment and decontamination of biological pathogens in the tested pilot-scale feed mill.