Antibacterial activity of egg yolk antibody (IgY) against Listeria monocytogenes and preliminary evaluation of its potential for food preservation

BACKGROUND: Egg yolk antibody (IgY) is a unique type of immunoglobulin found in egg yolks, and many reports have described its ability to inhibit corresponding antigen bacteria. The objective of this study was to evaluate the antibacterial activity of IgY specific to Listeria monocytogenes, an important food pathogen to both humans and animals, as well as its potential use for food preservation. RESULTS: Specific IgY was generated by immunising Leghorn chickens with whole cells of L. monocytogenes, and its inhibitory effect on bacterial growth was tested in liquid medium and food samples. After 8 h of incubation with specific IgY, there was a significant decrease in the growth (absorbance at 600 nm) of L. monocytogenes in comparison with controls. IgY also inhibited the growth of L. monocytogenes inoculated onto fresh or smoked salmon samples. Compared with those of blanks, numbers of L. monocytogenes were reduced by more than 2 log units after 15 days of storage at 6 ± 1 °C in the presence of specific IgY. CONCLUSION: The results suggest the potential application of specific IgY as a natural antimicrobial agent for food preservation. Copyright © 2011 Society of Chemical Industry     

Application of nisin,CO2 and a permeabilizing agent in the preservation of refrigerated blue whiting (Micromesistius poutassou)

Gram‐negative bacteria are mainly responsible for spoilage of refrigerated fish. Nevertheless, to preserve refrigerated fresh fish no additives are permitted and only packaging as in a modified atmosphere can be used. Despite the fact that the present‐day application of nisin has been extended to dairy and meat products, its ineffectiveness against Gram‐negative bacteria complicates application to fresh fish unless combined with other additives such as chelators. With this aim, agents that increase the permeability of the membrane and facilitate the diffusion of many hydrophobic antimicrobial agents were screened for their combined activity with nisin against different genera of Gram‐negative bacteria. Quantification of the synergistic effects has shown sodium hexamethaphosphate (SMP) and ethylenediaminetetraacetic acid (EDTA) were the most effective. Further studies were done on the combined effects of nisin, SMP and CO₂ in the preservation of fresh fish. A factorial experimental design was used to determine synergism between these variables. Logarithmic values of total viable counts and total volatile bases after 12 days of storage were described by empirical equations. Analysis of the results showed a marked effect of CO₂ in decreasing both variables. Whereas CO₂ interacted positively with nisin and SMP, a non‐significant interaction was observed between nisin and SMP. It is concluded that CO₂ interacts readily with SMP, and may offer an additional advantage for CO₂‐rich atmospheres in food preservation. Copyright © 2005 Society of Chemical Industry     

Quorum Sensing Involved in the Spoilage Process of the Skin and Flesh of Vacuum‐Packaged Farmed Turbot (Scophthalmus maximus) Stored at 4 °C

Fish skin has both positive and negative effects on the shelf‐life of the fish. This study aimed to investigate the spoilage process of the skin and flesh of refrigerated farmed turbot (Scophthalmus maximus) with vacuum packaging. Microbial community changes were analyzed by combining culture‐dependent and culture‐independent methods. The results indicated that the shelf‐life of vacuum‐packaged refrigerated turbot was 16 d; skin mucus was the interference factor of turbot quality. The culture‐dependent analysis demonstrated that the total viable counts and the population of H₂S‐producing bacteria, Pseudomonas spp., Enterobacteriaceae spp., and lactic acid bacteria in skin had a range of 0.45 to 1.40 log (CFU/g) higher than the microbial numbers in flesh after 16 d in storage. 16S high throughout sequencing results demonstrated that the compositions of spoilage microbes were similar in skin and flesh. Shewanella spp., followed by Carnobacterium spp., was the dominant spoilage organism at day 16. Quorum sensing (QS) signaling activity increased during the storage. Exogenous N‐butanoyl‐L‐homoserinelactone(C4‐HSL) and N‐hexanoyl‐Lhomoserine lactone (C6‐HSL) significantly accelerated the spoilage process of refrigerated turbot, while the addition of 4, 5‐Dihydroxypentane‐2, 3‐dione (DPD) prolonged the lag phase duration. Therefore, QS may be involved in the spoilage process of refrigerated turbot.     

Antimicrobial and antioxidant properties of burgers with quinoa peptide-loaded nanoliposomes

This study was aimed to examine the antioxidant (thiobarbituric acid reactive substances [TBARS], peroxide value [PV], and as well as antimicrobial (total bacterial count, Staphylococcus aureus [S. aureus], mold and yeast counts) activities and spoilage indices (total volatile base nitrogen [TVB-N]) of quinoa peptide-loaded liposomes incorporated into burger during 12 days of refrigerated storage. Among four prepared batches, the lowest TBARS, PV, and TVB-N values (0.281 ± 0.05 mg MDA/kg, 3.25 ± 0.34 mEq/kg and 18.65 ± 0.88%, respectively) were correlated with T3 treatment (5 mg/ml peptide). A significant different in the antimicrobial activity among each treatment after 12 days of storage was noted. At the end of refrigerated storage, the highest mean of total bacterial count, S. aureus, mold and yeast (8.36 ± 0.22, 4.19 ± 0.2, and 3.28 ± 0.05 log CFU/g, respectively) were observed in control group, while the lowest corresponded values (3.95 ± 0.2, 2.52 ± 0.25, and 1.52 ± 0.02 log CFU/g, respectively) were noted in T3 treatment. The results showed that the antimicrobial and antioxidant properties of the examined burgers were improved by incorporation of liposomes quinoa peptides encapsulated.     

Alginate–calcium coating incorporating nisin and EDTA maintains the quality of fresh northern snakehead (Channa argus) fillets stored at 4 °C

BACKGROUND: Antimicrobial packaging is a novel food packaging technology for controlling the growth of food‐borne pathogens or spoilage bacteria in ready‐to‐eat food products. Fresh fish are highly perishable foodstuffs and are extremely susceptible to microbial activities. An alginate–calcium coating incorporating nisin and ethylenediaminetetraacetic acid (EDTA) was used as an antimicrobial packaging to maintain the quality of northern snakehead (Channa argus) at refrigeration temperature (4 ± 1 °C). Northern snakehead fillets were left untreated (CK), or were treated with 1000 IU mL^?1 nisin and 150 µg mL^?1 EDTA (T1), alginate–calcium coating (T2), or alginate–calcium coating incorporating 1000 IU mL^?1 nisin and 150 µg mL^?1 EDTA (T3). RESULTS: All treatments retarded the decay of the fish fillets. T1 more efficiently inhibited the growth of total viable mesophilic bacteria (P < 0.05) and total psychrophilic bacteria (P > 0.05) than did T2 or T3. Coating treatments predominantly reduced chemical spoilage, reflected in total volatile base nitrogen (P > 0.05), trimethylamine (P > 0.05), pH (P < 0.05), and thiobarbituric acid (P < 0.05), retarded water loss (P < 0.05), and increased the overall sensory scores of fish fillets (P < 0.05) compared with those of T1. There was no significant difference between the coating treatments T2 and T3 (P > 0.05). CONCLUSION: Alginate–calcium coating treatments efficiently enhanced the quality of northern snakehead fillets during storage. Copyright © 2009 Society of Chemical Industry     

ANTIMICROBIAL SCREENING OF DECALEPIS HAMILTONII WIGHT AND ARN. (ASCLEPIADACEAE) ROOT EXTRACTS AGAINST FOOD-RELATED MICROORGANISMS

The antimicrobial activity of petroleum ether, benzene, chloroform, ethyl acetate and methanol extracts of Decalepis hamiltonii Wight and Arn. (Asclepiadaceae) against 15 different food‐related microorganisms was evaluated. Among the solvent extracts, methanolic and petroleum ether extracts are the most active exhibiting growth inhibition of 11 and nine of the 15 organisms, respectively. The inhibitory activity was comparable with the respective standards. The results suggest that methanolic and petroleum ether extracts can be used for the bioactivity‐guided isolation of novel antimicrobials which prevents food spoilage.     

Determination of volatile compounds of precooked prawn (Penaeus vannamei) and cultured gilthead sea bream (Sparus aurata) stored in ice as possible spoilage markers using solid phase microextraction and gas chromatography/mass spectrometry

BACKGROUND: Solid phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS) have been reported as useful techniques for analysing volatile compounds to monitor fish freshness. In this study, SPME/GC/MS was applied to cultured gilthead sea bream and precooked prawn stored in ice for 6 days in order to find possible markers of spoilage. Total volatile basic nitrogen (TVB‐N) was also determined as a common index of spoilage. RESULTS: The TVB‐N value at the end of the storage period for cultured gilthead sea bream (302.40 ± 8.50 mg kg^?1) was within the range of acceptability for edible fish (300–400 mg kg^?1) but could be considered at the beginning of spoilage. For precooked prawn the TVB‐N value at day 6 (863.04 ± 7.84 mg kg^?1) was not acceptable for human consumption. SPME/GC/MS identified 30 compounds in cultured gilthead sea bream and 49 compounds in precooked prawn. In particular, 3‐methyl‐1‐butanol, 2‐methylbutanal, 3‐methylbutanal and 3‐hydroxy‐2‐butanone increased during refrigerated storage both in the two species investigated here and in other species reported elsewhere and could be considered as markers of spoilage. CONCLUSION: This study confirmed that SPME/GC/MS can be considered an efficient method suitable for analysing the volatile compounds of both raw fish and fishery products in order to monitor loss of freshness. Copyright © 2008 Society of Chemical Industry     

The effectiveness of ozone and acidulant treatments in extending the refrigerated shelf life of fresh-cut potatoes

Abstract: The objective of the study was to determine the effectiveness of acidulant dip treatments (with or without aqueous ozone) to reduce enzymatic browning and to extend the shelf life of fresh‐cut potato slices during refrigerated storage (4 °C) for 28 d. Potato slices subjected to aqueous ozone (2 ppm) had significantly (P≤ 0.05) higher L‐values and lower a‐values, but ozone did not appear to have any effect on aerobic plate counts (APCs) or polyphenol oxidase (PPO) activity. NatureSeal (NS) and sodium acid sulfate (SAS) were the most effective acidulant treatments in reducing browning (significantly [P≤ 0.05] higher L‐values, lower a‐values, and browning index values) regardless of ozone treatment. NS and SAS also had lower PPO activity compared to other treatments on days 0 and 28, and significantly (P≤ 0.05) lower APCs (≤2.00 log CFU/g) over refrigerated storage. Therefore, the SAS treatment was comparable to NS, a commercially available product, and showed promise as an effective antibrowning dip to reduce browning and spoilage in fresh‐cut potato products. Practical Application: A 1% SAS dip treatment which included 1% citric and 1% ascorbic acid was found to be an effective antibrowning dip for fresh‐cut potatoes along with NatureSeal®'s PS‐10, compared to other treatments. They were both effective in maintaining low microbial counts over refrigerated storage. Additionally, aqueous ozone washes (2 ppm) showed significant benefits to reduce browning; however, ozone did not affect microbial counts or PPO enzyme activity. Therefore, the SAS treatment could have potential use in the fruit and vegetable industry to reduce browning and spoilage in fresh‐cut potato products.     

Microbiological aspects and shelf life of processed seafood products

BACKGROUND: Fresh fish and seafoods are very perishable products mainly owing to microbial activity of specific spoilage micro‐organisms. Application of hurdle technology leads to a variety of processed products with extended shelf life. In this study, sensory evaluation and microbiological analysis were carried out on 17 processed seafood products stored at 4 °C to determine their shelf life and the predominant spoilage micro‐organisms. RESULTS: Shelf life determined by sensory analysis varied from 66 to 180 days depending on the product. The cause of spoilage for most of the products was the development of off‐flavours/off‐odours, while two products were rejected owing to oil discolouration. Pseudomonads were in most cases below detection limit. H₂S‐producing bacteria, Brochothrix thermosphacta and Enterobacteriaceae were below detection limit throughout the experiment. The predominant spoilage micro‐organisms were lactic acid bacteria and yeasts. Hygiene indicators such as Staphylococcus spp. and total coliforms were also below detection limit in all samples. CONCLUSION: Primarily the initial pH and secondarily the NaCl content determined shelf life duration. Under the applied conditions, lactic acid bacteria and yeasts predominated. The contribution of chemical oxidation and/or autolysis to spoilage and shelf life might be important for most of the products. © 2012 Society of Chemical Industry     

Inhibitory activity of plumbagin produced by Drosera intermedia on food spoilage fungi

BACKGROUND: The aim of this study was to investigate the growth‐inhibiting efficacy of Drosera intermedia extracts (water, methanol and n‐hexane) against four food spoilage yeasts and five filamentous fungi strains responsible for food deterioration and associated with mycotoxin production, in order to identify potential antimycotic agents. RESULTS: The n‐hexane extract showed a broad activity spectrum against all tested microorganisms, followed, in activity, by the methanol and water extracts. The major component of the n‐hexane extract was purified using a solid‐phase extraction column and identified as plumbagin. Results show that high‐purity plumbagin can be produced from D. intermedia cultures following a simple and effective isolation procedure. A sample of purified plumbagin was tested against the same panel of microorganisms and high growth‐inhibiting capacity was observed. Minimum inhibitory concentrations less than 2 µg mL^?1 were obtained against the filamentous fungi. In the case of the species Aspergillus fumigatus, A. niger and A. flavus , activities comparable to miconazole were obtained. CONCLUSION: The results obtained provided evidence of the antimycotic activity of plumbagin, suggesting that D. intermedia could be the source of an interesting compound for the food industry as an alternative to preservatives. Copyright © 2011 Society of Chemical Industry     

Investigating the Antimicrobial Efficacy of a Lactococcal Bacteriocin for the Development of Microbiologically Stable Beer

Barley isolate Lactococcus lactis M30 produces an antimicrobial proteinaceous activity, which at least under laboratory conditions was shown to target beer spoiling lactic acid bacteria, including Lactobacillus brevis BSH9. The aim of this study was to investigate the application of this antibacterial activity at various stages of the brewing process and in packaged beer. Lactococcus lactis M30 was shown to produce the antimicrobial activity during growth under specific conditions in fortified unhopped wort. However, this activity was lost during wort boiling and yeast fermentation. When the bacteriocin was added directly to beer it retained in vitro activity following pasteurisation, while it was also shown to inhibit growth in situ when pasteurised beer was challenged with low levels of the beer spoiling Lactobacillus brevis BSH9 culture. The capacity of the bacteriocin to prevent microbial spoilage of beer was tested at various temperatures over a period of seven weeks. Storage of bacteriocin‐containing beer at 30°C or room temperature resulted in a decrease in antimicrobial activity over time, but when refrigerated or frozen, this beer retained sufficient activity to be effective against Lactobacillus brevis BSH9.     

The use of nisin as a preservative in fish sausage stored at ambient (28 ± 2 °C) and refrigerated (6 ± 2 °C) temperatures

Summary The efficacy of nisin at three different concentrations, 12.5, 25 and 50 ppm, on the keeping quality of fish sausage in synthetic casing at ambient (28 ± 2 °C) and refrigerated (6 ± 2 °C) temperatures was assessed. Gel strength, expressible water content, total volatile base nitrogen, total plate count and aerobic spore counts were affected by the storage temperatures and nisin concentrations used. Fish sausage treated with 50 ppm of nisin was acceptable after storage at ambient temperature for 20–22 days compared with the control, which were acceptable only for 2 days. The keeping quality of the sausages, at refrigerated temperature, varied from 30 days in the control to 150 days in 50‐ppm nisin‐treated samples. The residual nisin decreased slowly in samples stored at refrigerated temperature, whereas, in fish sausages stored at ambient temperature, the decrease was rapid. Nisin at 50‐ppm level showed a significant effect (P ≤ 0.05) on the gel strength and overall acceptability scores both at ambient and refrigerated temperature.     

Predictive models for bacterial growth in sea bass (Dicentrarchus labrax) stored in ice

The purpose of this paper was to estimate microbial growth through predictive modelling as a key element in determining the quantitative microbiological contamination of sea bass stored in ice and cultivated in different seasons of the year. In the present study, two different statistical models were used to analyse changes in microbial growth in whole, ungutted sea bass (Dicentrarchus labrax) stored in ice. The total counts of aerobic mesophilic and psychrotrophic bacteria, Pseudomonas sp., Aeromonas sp., Shewanella putrefaciens, Enterobacteriaceae, sulphide‐reducing Clostridium and Photobacterium phosphoreum were determined in muscle, skin and gills over an 18‐day period using traditional methods and evaluating the seasonal effect. The results showed that specific spoilage bacteria (SSB) were dominant in all tissues analysed, but were mainly found in the gills. Predictive modelling showed a seasonal effect among the fish analysed. The application of these models can contribute to the improvement of food safety control by improving knowledge of the microorganisms responsible for the spoilage and deterioration of sea bass.     

Microbial ecology of artisanal products from North West of Italy and antimicrobial activity of the autochthonous populations

A total of about 1000 autochthonous strains were isolated from fresh and fermented artisanal products (cheese and meat) of the Piedmont area (Northwest of Italy) and screened for their antimicrobial activity. Among them, 98 bacteriocin-producing strains were detected and identified by molecular methods and genetic determinants encoding the antimicrobial proteins were targeted by PCR. The bacteriocins produced by isolates showed antimicrobial activity towards different spoilage and pathogenic microorganisms such as Listeria monocytogenes, Staphylococcus aureus, Clostridium tyrobutyricum and Brochothrix thermosphacta. Identification of the bacteriocin producing strains resulted in a large number of Lactococcus and Enterococcus strains with a broad spectrum of activity towards more that one spoilage and/or pathogenic microorganisms tested. In several strains it was possible to detect a combination of different genes responsible for the bacteriocin activity. These newly isolated bacteriocin-producing strains represent potential starters or co-starters to be used in food stabilization using the approach of the biopreservation.     

Effect of Eryngium caeruleum essential oil on microbial and sensory quality of minced fish and fate of Listeria monocytogenes during the storage at 4°C

The present study investigated in vitro antimicrobial activity of Eryngium caeruleum essential oil (EEO) against five foodborne pathogenic bacteria based on microdilution and disk diffusion methods. Moreover, its effects on specific spoilage microorganisms, inoculated Listeria monocytogenes, and its sensory changes in minced fish were evaluated during 12 days of storage at refrigeration temperature. The results showed that Staphylococcus aureus and Escherichia coli were the most sensitive and the most resistant bacteria with a minimum inhibitory concentration of 0.125 and 1 mg/ml, as well as inhibition zones of 15.66 and 11.66 mm, respectively. Regarding the antimicrobial effect of EEO on the microbial profile and inoculated L. monocytogenes, treating with 0.4% EEO caused a significant decrease in the studied microorganisms when compared to the control group (p < 0.05). In addition, considering the sensory evaluation, the best scores were observed for the samples treated with 0.2% and 0.4% EEO. However, none of the groups obtained acceptable scores until the final day of storage except for the color attribute. In general, sensory evaluation and its correlation with microbial counting indicated that the treatment with 0.4% EEO was able to preserve the microbial quality of the minced fish at refrigeration temperature without any undesirable sensory effects.     

Effect of washing with tap and treated seawater on the quality of whole scad (<Emphasis Type="Italic">Trachurus trachurus</Emphasis>)

The effects of washing whole scad (Trachurus trachurus) with fresh water and seawater were evaluated. Fish evaluations for 12 days of storage in ice included sensory (EU and the QIM scheme), physical (Torrymeter and Freshmeter) and microbiological changes [counts of total bacterial numbers, specific spoilage organisms (SSO) and Pseudomonas]. The main conclusions are that both kinds of wash influenced physical and microbiological evaluations (mainly on Freshmeter values and counts of SSO). Washes seemed to interfere with properties of the fish skin cells that influenced the instrumental readings. By sensory means, the effects were not detected, resulting in similar shelf-lives, although some slight differences could be observed in mucus brightness in the first 2 days storage in ice. No significant differences were found between washing with tap water and treated seawater, but slightly lower values were obtained initially with seawater, in the first 24–48 h, with the Torrymeter and the Freshmeter.     

Growth Inhibitory Effect of Chicken Egg Yolk Antibody (IgY) on Escherichia coli O157:H7

Escherichia coli O157:H7‐specific antibodies (immunoglobulin Y [IgY]) were isolated by the water‐dilution method from the egg yolk of chickens that were immunized with E. coli O157:H7 whole cells. The specific‐binding activity of IgY against E. coli O157:H7 as determined by the enzyme immuno assay showed high levels of activity against bacterial whole cells. IgY binding activity was further demonstrated to have an inhibitory effect on E. coli O157:H7 growth in a liquid medium. The antibacterial function of IgY appeared to result from the interaction of IgY with surface components of E. coli O157:H7, as proven from observation of immunofluorescence and immunoelectron microscopy.     

Thymol and modified atmosphere packaging to control microbiological spoilage in packed fresh cod hamburgers

A study on the use of mild technologies to produce packaged fish hamburgers was presented. In particular, the antimicrobial effect of some natural compounds (carvacrol, eugenol, thymol, green tea extract, rosemary extract, grapefruit seed extract and lemon extract), at various concentrations (500–10 000 ppm), was screened in vitro against the main fish spoilage micro‐organisms (Shewanella putrefaciens and Photobacterium phosphoreum). Lemon extract and thymol, in combination with modified atmosphere packaging, showed the greatest inhibition activity, therefore, thymol was subsequently used as an ingredient for producing fish hamburgers. Results pointed out that this combination is effective in controlling the growth of microbial species mainly involved in fresh fish spoilage; in particular, it significantly (P < 0.05) reduced the growth rate of bacterial population, performing about 4.8 log CFU g^?1 and 6.5 log CFU g^?1 reduction of the hydrogen sulphide producing bacteria and psychrotrophic aerobic specific spoilage organisms cell load, respectively, if compared with the control.     

Development of a quality index method (QIM) sensory scheme and study of shelf-life of ice-stored blackspot seabream (Pagellus bogaraveo)

The aim of this work was to develop a quality index method (QIM) scheme for whole ice-boxed refrigerated blackspot seabream and to perform shelf-life evaluations, using sensory analysis, GR Torrymeter measurements and bacterial counts of specific spoilage organisms (SSO) during chilled storage. A QIM scheme based on a total of 30 demerit points was developed. Sensory, physical and microbiological data were integrated and used to determine the rejection point. Results indicated that the shelf-life of blackspot seabream is around 12–13 days.     

Antimicrobial activity of eucalyptus essential oil against Pseudomonas in vitro and potential application in refrigerated storage of pork meat

The objective of this study was to investigate the influence of eucalyptus essential oil (EO) on the Pseudomonas in vitro and application in the pork meat. Eucalyptus oil showed the greatest antimicrobial activity to Pseudomonas among five EOs (tea tree oil, eucalyptus oil, clove oil, mentha oil and garlic oil) in vitro. Eucalyptus oil treatment caused the increasing of membrane permeability and the modifications of ultrastructure and morphology of Pseudomonas spp. and P. aeruginosa. Treatment of refrigerated pork with eucalyptus oil led to a significant decrease P. spp. count, pH, total volatile basic nitrogen and 2‐thiobarbituric acid values, as well as higher sensory scores of custom acceptance than those in control at the end of cold storage. It was conclude that eucalyptus EO is a potential alternative to chemical treatment to control the spoilage and extend the shelf life of refrigerated pork meat.