Dual bacterial strains TTI for monitoring fish quality in food cold chain.

Most microbial time-temperature indicators (TTIs) considered only one spoilage strain. This research compared single and dual spoilage strains-based microbial TTI for quality changes of chilled grouper fish (Epinephelus fuscoguttatus x E. lanceolatus) fillet products during distribution. The next-generation sequencing (NGS) and traditional plate count approach showed that Pseudomonas fragi and Vibrio parahaemolyticus were specific spoilage bacteria at 7 and 15°C. A dual-strain TTI response provides more accurate results than a single-strain TTI and provides an irreversible color change from yellow to reddish-brown, showing levels of fish freshness. The microbial TTI comprises fish spoilage bacteria strains with 3 log CFU/ml, a nutrient broth supplemented with 2% NaCl as a medium, and phenol red with 0.25 mg/ml as a pH indicator. Overall, this study points to the applicability of a dual-strain microbial TTI as a valuable tool for monitoring fish quality changes during cold chain break condition.

Applications of Nisin and EDTA in Food Packaging for Improving Fabricated Chitosan-Polylactate Plastic Film Performance and Fish Fillet Preservation.

This study aimed to increase the antibacterial activity of chitosan-polylactic acid (PLA) composite film by adding nisin and ethylenediaminetetraacetic acid (EDTA). We evaluated the mechanical, physicochemical, and antibacterial properties of various PLA composite films, as well as the enhancement effect of PLA composite films with EDTA + nisin on the preservation of grouper fillets. Films of PLA alone, PLA plus chitosan (C5), PLA plus nisin + EDTA (EN2), and PLA plus chitosan plus nisin + EDTA (C5EN1 and C5EN2) were prepared. The addition of EDTA + nisin to the chitosan-PLA matrix significantly improved the antibacterial activity of the PLA composite film, with C5EN1 and C5EN2 films showing the highest antibacterial activity among the five films. Compared with the fish samples covered by C5, the counts of several microbial categories (i.e., mesophilic bacteria, psychrotrophic bacteria, coliforms, Aeromonas, Pseudomonas, and Vibrio) and total volatile basic nitrogen content in fish were significantly reduced in the samples covered by C5EN1. In addition, the counts of samples covered by C5EN1 or C5 were significantly lower compared to the uncovered and PLA film-covered samples.

Antibacterial Activity of Chitosan-Polylactate Fabricated Plastic Film and Its Application on the Preservation of Fish Fillet.

This research prepared chitosan-PLA plastic films by extrusion, analyzed the physical and mechanical properties and antibacterial activity of the fabricated plastic films, and used them to preserve grouper fillet. We added chitosan (220 kDa, 93% DD) in the weight ratio of 0.5-2% into the PLA to prepare the chitosan-PLA films. With the increasing chitosan dosage, both the water vapor transmission rate and moisture content of chitosan-PLA films increased. Among the three doses of chitosan (0.5%, 1%, and 2%) added to PLA, 0.5% chitosan-PLA film had the highest antibacterial activity. This plastic film had an inhibitory efficiency of over 95% against Escherichia coli, Pseudomonas fluorescens, and Staphylococcus aureus. The action of covering the fish fillet with 0.5% chitosan-PLA film significantly reduced several microbes' counting (i.e., mesophiles, psychrophiles, coliforms, Pseudomonas, Aeromonas, and Vibrio) and total volatile basic nitrogen (TVBN) value in the grouper fillets stored at 4 °C. Thus, such action prolongs the fish fillets' shelf life to up to at least nine days, and this 0.5% chitosan-PLA film shows promising potential for preserving refrigerated fish.

Suppressive effects of an apoptotic mimicry prepared from jumbo-flying squid-skin phospholipids on the osteoclastogenesis in receptor activator of nuclear factor kappa B ligand/macrophage colony-stimulating factor-induced RAW 264.7 cells.

BACKGROUND: Liposomes containing docosahexaenoic acid (DHA) and phosphatidylserine were claimed to inhibit osteoclast formation and bone resorption in the inflammatory status. Herein, we proposed that an apoptotic mimicry (SQ liposome) prepared from squid-skin phospholipids can explore the suppressive osteoclastogenesis. METHODS: The intermolecular fatty-acid composition in the phospholipid of squid-skin extract was analyzed by GC-FID. The SQ liposome structure was characterized by size distribution and zeta potential (ζ). RAW 264.7 cell is used to study the effect of SQ liposomes on osteoclast differentiation. Secretion of prostaglandin E2 (PGE2) and transforming growth factor-ß (TGF-ß) from RAW 264.7 cells were assayed. Antiosteoclastogenesis effects were performed via the tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cell (MNC) counting, bone resorption pit assay, and TRAP activity analysis. The specific gene expressions related to antiosteoclastogenesis were also detected. RESULTS: An apoptotic mimicry through the use of a single-layer liposome (SQ liposome) with phosphatidylserine exposure contains DHA (28.7%) and eicosapentaenoic acid (EPA, 11.8%). Co-treatment with receptor activator of nuclear factor kappa B ligand (RANKL)/macrophage colony-stimulating factor induced RAW 264.7-cell differentiation into mature osteoclasts, thus enhancing PGE2 and TGF-ß secretion. However, cotreatment with 1 mg/mL of SQ liposome restored (p < 0.05) the cell viabilities under the RANKL stress. Increased PGE2 levels was downregulated (p < 0.05) in cotreatments with 0.11 and 0.33 mg/mL of SQ liposome, but on the TGF-ß levels were not (p > 0.05) influenced in SQ liposome cotreatments. Cotreatments with 0.33-1 mg/mL of SQ liposome suppressed (p < 0.05) the osteoclast maturation (such as decreased MNCs and bone pit formation), inhibited TRAP activities, and downregulated the osteoclastogenesis-related gene expressions. CONCLUSION: In summary, current data support that a possible prevention of our prepared SQ liposomes which are rich in DHA and EPA on bone loss is through the suppression of osteoclastogenesis. Moreover, based on the results from this study an in vivo study warrants a further investigation.

Manufacture and characterization of anti-inflammatory liposomes from jumbo flying squid (Dosidicus gigas) skin phospholipid extraction.

The anti-inflammation properties of marine phospholipids enriched with n-3 fatty acids contribute to anti-inflammatory and inflammation-resolving mediators. Functional squid-skin (SQ) liposomes were manufactured from squid-skin phospholipids, and their anti-inflammatory effects were investigated. SQ liposomes included phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylcholine (PC), and lysophosphatidylcholine (Lyso-PC), and had an approximate diameter of 100 mm. When RAW264.7 cells were treated with the SQ liposome, no (p > 0.05) cytotoxicity was observed below a concentration of 7.5 mg mL-1. An SQ-liposome pretreatment of lipopolysaccharide (LPS)-induced RAW 264.7 cells showed decreased (p < 0.05) prostaglandin E2 (PGE2), nitric oxide (NO), interleukin-1beta (IL-1ß), IL-6, and tumor necrosis factor-alpha (TNF-α). The engulfment of SQ liposomes by the RAW264.7 cells resulted in lower (p < 0.05) LPS-induced intracellular levels of reactive oxygen species. Furthermore, an SQ-liposome administration ameliorated (p < 0.05) carrageenan-induced paw edema in mice. SQ liposomes may act via apoptotic mimicry to elicit the resolution of inflammation and prevent chronic inflammation-related diseases.

Effects of sizes and conformations of fish-scale collagen peptides on facial skin qualities and transdermal penetration efficiency.

Fish-scale collagen peptides (FSCPs) were prepared using a given combination of proteases to hydrolyze tilapia (Oreochromis sp.) scales. FSCPs were determined to stimulate fibroblast cells proliferation and procollagen synthesis in a time- and dose-dependent manner. The transdermal penetration capabilities of the fractionationed FSCPs were evaluated using the Franz-type diffusion cell model. The heavier FSCPs, 3500 and 4500 Da, showed higher cumulative penetration capability as opposed to the lighter FSCPs, 2000 and 1300 Da. In addition, the heavier seemed to preserve favorable coiled structures comparing to the lighter that presents mainly as linear under confocal scanning laser microscopy. FSCPs, particularly the heavier, were concluded to efficiently penetrate stratum corneum to epidermis and dermis, activate fibroblasts, and accelerate collagen synthesis. The heavier outweighs the lighter in transdermal penetration likely as a result of preserving the given desired structure feature.