Effects of curcumin-mediated photodynamic treatment on lipid degradation of oysters during refrigerated storage.

BACKGROUND: Oyster's lipid degradation leads to a decrease in edible and nutritional value. Curcumin-mediated photodynamic treatment (PDT) is an innovative non-thermal technology, although evaluation of the oyster's lipid degradation has been scarce. In the present study, we investigated peroxide value, thiobarbituric acid reactive substance, triacylglycerol and free fatty acids to evaluate the effect of curcumin-mediated PDT on lipid degradation of oysters during refrigerated storage. RESULTS: The results showed that curcumin-mediated PDT could delay oyster's lipid degradation. Next, the activities of enzymes were detected to determine the mechanisms behind the effects of curcumin-mediated PDT. It was revealed that the activities of lipase, phospholipase A2 (PLA2 ), phospholipase C (PLC), phospholipase D (PLD) and lipoxygenase (LOX) were significantly inhibited after curcumin-mediated PDT (P < 0.05). Furthermore, 16 s rRNA analysis established that the relative abundances of Pseudoalteromonas and Psychrilyobacter were reduced by 51.58% and 43.82%, respectively, after curcumin-mediated PDT. CONCLUSION: Curcumin-mediated PDT could delay oyster's lipid degradation by inhibiting the activities of lipase, PLA2 , PLC, PLD and LOX, as well as by changing the oyster's microbial composition, reducing the relative abundance of Pseudoalteromonas and Psychrilyobacter. © 2021 Society of Chemical Industry.

Photodynamic effect of curcumin on Vibrio parahaemolyticus.

Vibrio parahaemolyticus (V. parahaemolyticus) is currently a major cause of bacterial diarrhoea associated with seafood consumption. The objective of this study was to determine the inactivation effect of curcumin-mediated photodynamic action on V. parahaemolyticus. First of all, V. parahaemolyticus suspended in PBS buffer was irradiated by a visible light from a LED light source with an energy density of 3.6J/cm(2). Colony forming units (CFU) were counted and the viability of V. parahaemolyticus cells was calculated after treatment. Singlet oxygen ((1)O2) production after photodynamic action of curcumin was evaluated using 9,10-Anthracenediyl-bis (methylene) dimalonic acid (ADMA). Bacterial outer membrane protein was extracted and analyzed using electrophoresis SDS-PAGE. DNA and RNA of V. parahaemolyticus were also extracted and analyzed using agarose gel electrophoresis after photodynamic treatment. Finally, the efficacy of photodynamic action of curcumin was preliminarily evaluated in the decontamination of V. parahaemolyticus in oyster. Results showed that the viability of V. parahaemolyticus was significantly decreased to non-detectable levels over 6.5-log reductions with the curcumin concentration of 10 and 20µM. Photodynamic action of curcumin significantly increased the singlet oxygen level with the curcumin concentration of 10µM. Notable damage was found to bacterial outer membrane proteins and genetic materials after photodynamic treatment. Photodynamic action of curcumin reduced the number of V. parahaemolyticus contaminating in oyster to non-detectable level. Our findings demonstrated that photodynamic action of curcumin could be a potentially good method to inactivate Vibrio parahaemolyticus contaminating in oyster.

Virus, protozoa and organic compounds decay in depurated oysters.

AIMS: (1) Evaluate the dynamic of the depuration process of Crassostrea gigas oysters using different ultraviolet doses with different amounts of contaminants (virus, protozoa and organic contaminants) and (2) investigate the morphological changes in the oysters' tissues produced by the depuration procedures. METHODS: The oysters were allocated in sites with different degrees of contamination and analyzed after 14 days. Some animals were used as positive controls by artificial bioaccumulation with HAdV2 and MNV1 and subjected to depuration assays using UV lamps (18 or 36 W) for 168 h. The following pollutants were researched in the naturally contaminated oysters, oysters after 14 days in sites and oysters during the depuration processes: virus (HAdV, HAV, HuNoV GI/GII and JCPyV), by (RT) qPCR; protozoa (Cryptosporidium and Giardia species), by immunomagnetic separation and immunofluorescence; and organic compounds (AHs, PAHs, LABs, PCBs and organochlorine pesticides-OCs), by chromatography. Changes in the oysters' tissues produced by the depuration processes were also evaluated using histochemical analysis by light microscopy. In the artificially bioaccumulated oysters, only HAdV2 and MNV1 were investigated by (RT) qPCR before the depuration procedures and after 96 and 168 h of these procedures. RESULTS: At 14 days post-allocation, HAdV was found in all the sites (6.2 × 105 to 4.4 × 107 GC g(-1)), and Giardia species in only one site. Levels of PCBs and OCs in the oyster's tissues were below the detection limit for all samples. AHs (3.5 to 4.4 µg g(-1)), PAHs (11 to 191 ng g(-1)) and LABs (57 to 751 ng g(-1)) were detected in the samples from 3 sites. During the depuration assays, we found HAdV, Giardia and Cryptosporidium species until 168 h, independent of UV treatment. AHs, PAHs and LABs were found also after 168 h of depuration (36 W and without UV lamp). The depuration procedures did not produce changes in the oysters' tissues. In the artificially contaminated and depurated oysters, we detected HAdV until 168 h and MNV1 until 96 h of depuration. CONCLUSION: The applied depuration treatments were unable to eliminate the protozoa or to degrade the HAdV genomes but were able to degrade the MNV1 genomes. Similarly, the UV water treatment was not efficient for aliphatic hydrocarbons, PAHs and LABs, as their concentrations were equivalent or higher to the concentrations of the control samples and samples from depuration tanks without UV treatment.

Feasibility studies into the production of gamma-irradiated oyster tissue reference materials for paralytic shellfish poisoning toxins.

A study was conducted to assess the feasibility for the production of sterile, stable and homogenous shellfish reference materials containing known concentrations of paralytic shellfish poisoning (PSP) toxins. Pacific oysters were contaminated with toxins following mass culturing of toxic algae and shellfish feeding experiments. Live oysters were shucked and tissues homogenised, before measuring into multiple aliquots, with one batch subjected to gamma irradiation treatment and the other remaining untreated. The homogeneity of both batches of samples was assessed using a pre-column oxidation liquid chromatography with fluorescence detection (Pre-COX LC-FLD) method and shown to be within the limits of normal within-batch repeatability. A twelve-month stability experiment was conducted for both untreated and gamma irradiated batches, specifically examining the effects of long term storage at -20 °C, +4 °C and +40 °C. Results indicated mostly good stability of PSP toxins in both materials when stored frozen at -20 °C, but with the instability of GTX2&3 concentrations in the untreated tissues eliminated in the irradiated tissues. Analysis using a post-column oxidation (PCOX) LC-FLD method also showed epimerisation in both GTX1&4 and GTX2&3 epimeric pairs in untreated samples after only 6 months frozen storage. This issue was not present in the tissues irradiated before long term storage. Biological activity testing confirmed the absence of bacteria in the irradiated samples throughout the 12 month study period. With such results there was clear evidence for the potential of increasing the scale of the mass culturing and shellfish feeding for the production of large batches of tissue suitable for the preparation of a certified matrix reference material. Overall results demonstrated the feasibility for production of oyster reference materials for PSTs, with evidence for prolonged stability following gamma irradiation treatment and storage at -20 °C.

Identifying irradiated oysters by luminescence techniques (TL & PSL).

This study shows an exhaustive comparison of different methods, based on luminescence techniques, to identify X-ray irradiated oysters at five different dose levels in the range 0.1-2 kGy and suggests a simple, fast and sequential routine analysis protocol. A total number of 50 oysters from North Sea, including 10 control samples, were analysed by using two photo-stimulated luminescence (PSL) methods (named A and B) and three thermoluminescence (TL) methods (named C, D and E), either on constituents or contaminating minerals from shells and intestines. Setting a lower threshold value T1 (1000 counts/60s) all of control specimens gave negative screening results while photo counts from irradiated samples were found to be higher than upper threshold T2 (4000 counts/60s), except at the lowest dose level 0.1 kGy for procedure A. All PSL calibrated analysis were successful and a sensitivity index was also determined to better classify obtained data according to the revised European Standard (EN 13751:2009). TL ratios, Glow 1 over second Glow 2, the latter after irradiating at 1 kGy and remeasuring the same minerals for each sample, showed values less than 0.1 related to untreated samples or higher than 0.1 for irradiated ones. Reported procedures were also tested over 60 days, longer than oyster shelf life confirming the applicability and feasibility of the proposed methods.

Thermoluminescence analysis of irradiated oyster shells.

This paper reports the thermoluminescence (TL) analysis performed on the oyster shells powder. TL response of (60)Co gamma-rays irradiated samples were studied in the range from 80 Gy to 8 kGy doses. TL signal of irradiated shell powder was higher as compared to the unirradiated control samples, which allowed to identify the irradiated oysters. Results show that the oyster shells have good TL properties and can be useful for the identification of irradiated seafood as well as for the evaluation of the treatment dose.

Dose characterization of the rad source 2400 X-ray irradiator for oyster pasteurization.

The RS 2400's cylindrical X-ray source yields dose rates high enough to allow the irradiator to replace widely used gamma irradiators. Except for the leftmost 5 cm, beam uniformity is within 10% at the tube surface. At maximum operating parameters, the beam has HVL(1)=13.66 mm aluminum, HC=0.47, and hv(eq)=88.5 keV. Maximum dose rates to tissue are 65 Gy min(-1)+/-3.1% at tube surface, 37 Gy min(-1)+/-3.1% at center of canisters, 14.1 Gy min(-1)+/-6.5% for thin-shelled oysters, and 12.3 Gy min(-1)+/-6.2% for thick-shelled oysters.

A continuous microwave system for prevention of invasive species during de-ballasting operation--death kinetics.

A continuous microwave heating system was tested for its effectiveness at removing potentially invasive organisms during deballasting operations. Four different organisms, namely Nannochloropsis oculata (microalgae), Artemia nauplii, Artemia adults and Crassosstrea virginica (oyster larvae) normally found in ballast water were investigated in a controlled study to quantify their survival after continuous microwave heating of synthetic ballast water. The experiments were performed in the microwave system using a 2 x 2 factorial design with power (2.5 and 4.5 kW) and flow rate (1.0 and 2.0 lpm) and the organisms subsequently subjected to different holding times. The control treatment was performed in a water bath using the same temperatures and holding times as in the case of the microwave treatment. Overall, the results obtained indicated that the microwave system was more effective in eliminating the organisms when compared with the control treatment. In most cases there were no survivors present after the microwave treatment at holding times above 100 s, and temperatures as low as 50 degrees C particularly for oyster larvae and Artemia adults. The results are promising, indicating that this technology has the potential to be an effective tool in controlling/preventing the introduction of invasive species into native environments.

Induction of haploid androgenesis in Pacific oyster by UV irradiation.

Androgenesis, development from paternal but not maternal chromosomes, can be induced in some organisms including fish, but has not been induced previously in mollusk. In this study we investigated the induction of haploid androgenesis in the Pacific oyster by ultraviolet irradiation and observed nuclear behavior in the androgenetic eggs. Irradiation for 90 seconds at a UV intensity of 72 erg/mm2 per second (6480 erg/mm2) was the optimal dose to achieve haploid androgenesis. The fertilization and development rates of D-shaped larvae decreased with increasing exposure time, and the development of the genetically inactivated eggs terminated before reaching the D-shaped stage. Cytologic observations showed that UV irradiation did not affect germinal vesicle breakdown or chromosomal condensation but caused various nuclear behavioral patterns during meiosis and first mitosis: 21.7% of eggs extruded all maternal chromosomes as 2 or 3 polar bodies, and 59.1% of eggs formed one female pronucleus. The maternally derived nucleus did not participate, or partially participated, in the first karyokinesis. The cytologic evidence demonstrates that the male genome is directing development in haploids produced by UV irradiation.

Inactivation by ionizing radiation of Salmonella enteritidis, Salmonella infantis, and Vibrio parahaemolyticus in oysters (Crassostrea brasiliana).

Irradiation is considered one of the most efficient technological processes for the reduction of microorganisms in food. It can be used to improve the safety of food products, and to extend their shelf lives. Oysters are considered one of the most important vehicles for pathogenic bacteria because of their feeding characteristics. The aim of this study was to evaluate the influence of a gamma radiation process on high levels of Salmonella Enteritidis, Salmonella Infantis, and Vibrio parahaemolyticus incorporated by oysters (Crassostrea brasiliana), as well as the effects of the process on the survival of the oysters and on their sensory attributes. The oysters were exposed to gamma radiation (60Co) in doses ranging from 0.5 to 3.0 kGy. A dose of 3.0 kGy was generally sufficient to reduce the level of Salmonella serotypes by 5 to 6 log10 units. A dose of 1.0 kGy was sufficient to produce a 6-log10 reduction in the level of V. parahaemolyticus. The highest irradiation dose did not kill the oysters or affect their sensory attributes. Hence, a dose of 3.0 kGy can be considered effective in inactivating Salmonella and V. parahaemolyticus in oysters without changing their odor, flavor, or appearance.

Multifrequency EPR study of carbonate- and sulfate-derived radicals produced by radiation in shells and corallite.

Shells of two sea mollusks (Venus sp.), pearl oyster (Meleagrina vulgaris) and corallite (white coral) were exposed to ionizing radiation (gamma and X rays) and then examined by EPR spectroscopy in X, Q and W band. The resulting spectra were analyzed and the g values of the EPR lines in the multicomponent spectra were determined. The increased resolution in Q- and W-band spectra allowed us to assign the observed lines to CO(2)(-) ion radicals (isotropic and orthorhombic), SO(2)(-) isotropic, SO(3)(-) (isotropic and axial), and Mn(2+) species. The assignments were confirmed by simulations of the spectra. Practical implications for the use of Q and/or W band in low-dose quantitative EPR measurements for dating and for accidental dose estimation are discussed.

Persistence of Vibrio vulnificus in tissues of Gulf Coast oysters, Crassostrea virginica, exposed to seawater disinfected with UV light.

Vibrio vulnificus is an estuarine bacterium which can cause opportunistic infections in humans consuming raw Gulf Coast oysters, Crassostrea virginica. Although V. vulnificus is known as a ubiquitous organism in the Gulf of Mexico, its ecological relationship with C. virginica has not been adequately defined. The objective of the present study was to test the hypothesis that V. vulnificus is a persistent microbial flora of oysters and unamenable to traditional methods of controlled purification, such as UV light depuration. Experimental depuration systems consisted of aquaria containing temperature-controlled seawater treated with UV light and 0.2-microns-pore-size filtration. V. vulnificus was enumerated in seawater, oyster shell biofilms, homogenates of whole oyster meats, and tissues including the hemolymph, digestive region, gills, mantle, and adductor muscle. Results showed that depuration systems conducted at temperatures greater than 23 degrees C caused V. vulnificus counts to increase in oysters, especially in the hemolymph, adductor muscle, and mantle. Throughout the process, depuration water contained high concentrations of V. vulnificus, indicating that the disinfection properties of UV radiation and 0.2-microns-pore-size filtration were less than the rate at which V. vulnificus was released into seawater. Approximately 10(5) to 10(6) V. vulnificus organisms were released from each oyster per hour, with 0.05 to 35% originating from shell surfaces. These surfaces contained greater than 10(3) V. vulnificus organisms per cm2. In contrast, when depuration seawater was maintained at 15 degrees C, V. vulnificus was not detected in seawater and multiplication in oyster tissues was inhibited.

Microbial flora of Pacific oysters (Crassostrea gigas) subjected to ultraviolet-irradiated seawater.

The ability of oysters to purge themselves of microbial contaminants was investigated by identifying the microorganisms retained by oysters after they have been subjected to ultraviolet (UV) light-treated seawater. A UV intensity of 960 muw per min per cm(2) reduced the microbial count of seawater from 263 to 13 per ml. The coliform multitube test (MPN) was reduced from a high of 17 to <0.18 per 100 ml. Over 75% of the microorganisms found in treated seawater were Acinetobacter/Moraxella, Vibrio/Pseudomonas type II, and Flavobacterium/Cytophaga. With the exception of coliforms, the microbial composition of oysters subjected to UV-treated seawater remained at levels comparable to the control oysters held in untreated seawater. Total counts ranged between 10(3) and 10(5)/g. The microorganism most frequently encountered were Flavobacterium/Cytophaga, Vibrio/Pseudomonas type II, Pseudomonas type III or IV, Acinetobacter/Moraxella, gram-positive cocci and Bacillus. Together they comprised over 90% of the flora. Coagulase-positive, deoxyribonuclease-positive, and beta-hemolytic cocci were found in some samples, as were V. parahaemolyticus, V. aliginolyticus, and Aeromonas species.