Occurrence and diversity of Listeria spp. in seafood processing plant environments

Listeria contamination in processing plant environments is a major issue for the seafood industry worldwide; faster and more reliable results are therefore desired for early detection and monitoring of environmental Listeria spp. This study aimed to gain a better understanding of the prevalence and diversity of Listeria spp., and to evaluate a rapid detection method, the 3M Molecular Detection Assay (MDA) Listeria, for its ability to detect Listeria spp. in environmental samples from seafood processing plants. Duplicate environmental sponge samples (n = 444) were collected from 152 different sites within three seafood processing plants, and analyzed for Listeria spp. by the MDA method (after 26 and 48 h of enrichment) and the U.S. Food and Drug Administration Bacteriological Analytical Manual method. Overall, detection of Listeria spp. by the two methods did not differ significantly (p > 0.05); 11 (4.9%) and 13 (5.9%) samples were positive for Listeria spp. by the MDA and FDA-BAM method, respectively. The sensitivity of the MDS was 87.0% (95% CI: 77.4–96.6%), specificity was 97.6% (95% CI: 95.5–99.7%), accuracy was 95.3%, and the positive predictive value was 89.4% (95% CI: 80.5–98.2%). Classification of 19 Listeria isolates by partial SigB sequencing analysis identified three allelic types. Twelve of these isolates were ATs 58 and 60 which were classified as Listeria monocytogenes lineage I and serotypes 1/2b, 3b, 4b, 4d, 4e, by multiplex-PCR serotyping. Six Listeria isolates were classified as Listeria innocua (AT31). Our data show that the 3M Molecular Detection Assay Listeria provides rapid and reliable results for detection and monitoring of Listeria spp., which are important for seafood processing plants. Effective Listeria monitoring programs will allow for improved development of Listeria control measures in order to minimize cross-contamination in finished products.     

Spatio-temporal variability in the fatty acid profile of the adductor muscle of the common cockle Cerastoderma edule and its relevance for tracing geographic origin

The present study aimed to identify the potential existence of spatio-temporal variability in the fatty acid (FA) profile of the adductor muscle (AM) of a commercially important bivalve, the common cockle Cerastoderma edule, and determine if such variability can be used to trace their geographic origin post-harvesting. Common cockles were sampled in eight ecosystems along the coast of mainland Portugal, as well as in two different channels within one of those ecosystems over two consecutive years. Results showed significant differences in FA profiles among ecosystems, namely due to different levels of eicosapentaenoic acid (EPA; 20:5n-3), arachidonic acid (20:4n-6), ratio of polyunsaturated FA (PUFA) n-3/n-6, monounsaturated FA (MUFA; 18:1n-9 and 20:1n-9/11) and bacterial FA (15:0, 17:0 and 18:1n-7). FA profiles also displayed significant differences between two consecutive years in channels from the same ecosystem. Overall, while the FA profile displayed by the AM of C. edule can be successfully used to trace their geographic origin, the existence of temporal variability requires a periodical verification of FA signatures to identify potential shifts, namely when comparing specimens from the same ecosystem.     

Italian market fish species identification and commercial frauds revealing by DNA sequencing

To guarantee consumers about the security of what they eat is the main goal of all the institutions working to assure food safety. In seafood field it appears of prime importance to be able to identify fish species throughout the production cycle also when the product has been already processed (breaded fish, fillets etc). In this context the latest system proposed for species identification is the so called “DNA-Barcoding”, made by carrying out the DNA sequencing of a standard barcode region. In the present study we carried out bi-directional sequence analysis of mitochondrial cytochrome b gene (Cytb) and cytochrome oxidase subunit I gene (COI). The main goal of the research was to unambiguously classify the most common fishes traded in Italy, being able to ascertain possible labeling frauds made substituting value species with less precious ones. With both genes we could correctly identify almost all the samples in study: respectively 58/58 with COI and 56/58 with Cytb. This study confirms mitochondrial genes Cytb and COI as good candidates for fish species identification by DNA sequencing. This method appears particularly suitable especially when morphological characterization is difficult, (for very close species), or impossible (for transformed foodstuffs) to carry out. What's more we could efficiently identify many fishes belonging to the Mediterranean fauna, which, to our knowledge, underwent for the first time this kind of analysis. Further development will concern the extension of the analysis to a largest number of local species.     

Development of lateral flow immunoassay for rapid control and quantification of the presence of the colorant Sudan I in spices and seafood

The lateral flow immunoassay for colorant Sudan I detection was developed using the monoclonal antibody to Sudan I conjugated with gold nanoparticles. Conditions of the assay were optimized for both qualitative control and quantification of Sudan I in food matrixes with minimal dilution of the extracting media (final content of methanol – 50%). In the case of photometric registration, the limit of Sudan I detection in extracts was 2.5 ng/mL (which corresponds to 10 μg/kg of food and spices), and the working range of determined concentrations was 13–240 ng/mL. The average error of measurements varied from 5.5 to 16.9%. The visual limit of detection, which is the minimum concentration causing absence of visible coloration at the test zone, was 1 μg/mL. The assay efficiency was confirmed by its application to determine Sudan I content in highly colored spices (turmeric, curry) and seafood (caviar, mussels, fish). Thus, the developed assay is suitable as a rapid, inexpensive tool for revealing Sudan I quantification by mobile laboratories and in out-of-laboratory conditions.     

Labelling accuracy in Tasmanian seafood: An investigation using DNA barcoding

Mislabelling of seafood products has been documented in numerous countries for over three-quarters of a century. With a trend towards increased consumption of seafood, the informed consumer demands accurately labelled products that provide full disclosure of composition. DNA barcoding can be used to accurately identify a seafood product to species based on its genetic signature, and so provides a means to test the authenticity and accuracy of seafood labelling. This can be especially useful for products such as fillets which have few or no unambiguous identifying characters, and can easily be mislabelled. We investigated labelling accuracy in seafood retailers in Tasmania, Australia. Thirty-eight seafood products were obtained from seafood retailers, sequenced for the barcoding gene region cytochrome oxidase subunit 1(CO1), and subsequently identified to species level by querying GenBank and Barcode of Life Data Systems (BOLD) DNA sequence records. Results were compared with standard fish names (SFN) prescribed under the Australian Fish Names Standard (AFNS) and FishBase. Of the 38 samples, none were deemed to have been mislabelled under Australian regulation, although in some cases naming discrepancies and ambiguity may cause confusion for some consumers. Our work, while reflecting high standards in Tasmanian seafood, highlights the need for mandatory standard labelling across all seafood products so as to eliminate any possible misrepresentation.     

A bacteriocin-like substance produced from Lactobacillus pentosus 39 is a natural antagonist for the control of Aeromonas hydrophila and Listeria monocytogenes in fresh salmon fillets

We studied the ability of Lactobacillus pentosus 39, a BLS (Bacteriocin-like substance)-producing strain, to control the growth of Aeromonas hydrophila ATCC 14715 and Listeria monocytogenes ATCC 19117 artificially added to fresh salmon fillets at refrigeration temperatures and under simulated cold-chain break conditions.At refrigeration temperatures, Lb. pentosus 39 protective culture and its putative bacteriocin significantly reduced A. hydrophila counts compared with the control (2.1 and 1.4 log CFU/g reductions, respectively). Similar behaviour was observed for L. monocytogenes (3.6 and 1.3 log CFU/g reductions, respectively).Under simulated cold-chain break conditions, an increase in temperature (30°C for 12h) produced an evident increase in the development of A. hydrophila, L. monocytogenes, but also of Lb. pentosus 39, with a consequent increase in BLS production. This condition resulted in a greater reduction of both pathogens compared with samples stored at 4°C throughout the experiment (2.8 log CFU/g reduction for A. hydrophila, 5.8 log CFU/g reduction for L. monocytogenes). In samples treated with the putative bacteriocin alone, a less marked decrease was observed.Our study demonstrates the capability of Lb. pentosus 39 to control the growth of psychrotrophic bacteria in an experimental seafood model system. A similar biopreservation technology could provide more prolonged shelf-life during storage of ready-to-eat seafood, ensuring safety, even under extreme conditions.     

Some Ozone Applications in Seafood

The use of ozonized seawater to reduce and eliminate bacterial pathogens in mariculture facilities and to extend shelf life of marine food products is demonstrated. Consequent benefits of this treatment are also discussed. Laboratory and pilot experiments were conducted using ozone gas to reduce disease-producing Vibrio sp. bacteria at a shrimp (Litopenaeus vannameii) hatchery in Ecuador, South America. Pacific Ocean seawater was treated in a 1,540 L capacity fiberglass contact tower (5–7 min retention) with an ozone oxidant residual of 0.07 mg/L. Prior to ozone treatment, Vibrio determined by TCBS plating was too numerous to count, causing shrimp to die of disease (30 tanks of 13,000 L each). After treatment, Vibrio counts and shrimp disease were eliminated, ozonized seawater decreased the time required for normal molting of shrimp and the total growth cycle was reduced by three days versus control water. From June 1991 until September 1992, survival rates of larval shrimp were robust, routine antibiotic addition was reduced, and one additional growth cycle was realized. Ozonized ice (fresh water) was prepared in the Milford Laboratory CT, USA; Gloucester Food Tech Lab, MA, USA; and a field station (brine water) for sockeye salmon (Oncorhynchus nerka) in Homer, AK. USA. In these studies, squid (Loligo pealei) and commercially captured salmon demonstrated a reduction in spoilage and extension of shelf life of 3 to 5 days' time using ozonized ice. Bacteria associated with commercial ice-producing machines were reduced by 4 logs using ozone treatment. In addition, no flavor aberration was noted using ozonized ice.     

Seafood safety and quality: An analysis of the supply chain in the Sultanate of Oman

The globalization of trade in fish has created many challenges for the developing world specifically with regard to food safety and quality. International organisations have established a good basis for standards in international trade. Whilst these requirements are frequently embraced by the major importers (such as Japan, the EU and the USA), they often impose additional safety requirements and regularly identify batches which fail to meet their strict standards. Creating an effective national seafood control system which meets both the internal national needs as well the requirements for the export market can be challenging. Many countries adopt a dual system where seafood products for the major export markets are subject to tight control whilst the majority of the products (whether for the local market or for more regional trade) are less tightly controlled. With regional liberalization also occurring, deciding on appropriate controls is complex. In the Sultanate of Oman, fisheries production is one of the countries' chief sources of economic revenue after oil production and is a major source of the national food supply. In this paper the structure of the fish supply chain has been analysed and highlighted the different routes operating for the different markets. Although much of the fish are consumed within Oman, there is a major export trade to the local regional markets. Much smaller quantities meet the more stringent standards imposed by the major importing countries and exports to these are limited. The paper has considered the development of the Omani fish control system including the key legislative documents and the administrative structures that have been developed. Establishing modern controls which satisfy the demands of the major importers is possible but places additional costs on businesses. Enhanced controls such as HACCP and other management standards are required but can be difficult to justify when alternative markets do not specify these. These enhanced controls do however provide additional consumer protection and can bring benefits to local consumers. The Omani government is attempting to upgrade the system of controls and has made tremendous progress toward the implementation of HACCP and introducing enhanced management systems into its industrial sector. The existence of strengthened legislative and government support, including subsidies, has encouraged some businesses to implement HACCP. The current control systems have been reviewed and an SWOT analysis approach used to identify key factors for their future development. The study shows that seafood products in the supply chain are often exposed to lengthy handling and distribution process before reaching the consumers, a typical issue faced by many developing countries. As seafood products are often perishable, they safety is compromised if not adequately controlled. The enforcement of current food safety laws in the Sultanate of Oman is shared across various government agencies. Consequently, there is a need to harmonize all regulatory requirements, enhancing the domestic food protection and to continue to work towards a fully risk-based approach in order to compete successfully in the global market.     

生吃水产品常见病原性生物污染的研究

为了解广东省生吃水产品以及生吃水产品养殖水等中常见病原性生物的污染情况，从1999年6月至2000年12月，对广东省18家酒店的生食生产品、养殖水和砧板进行卫生检验。结果从325份样品中，检出常见病原菌79株，包括7种致病菌，检出阳性率为24．31％，其中罗氏虾的阳性率高达50％。另外25份生鱼肉中的5份检出华支睾吸虫，检出率为20％。结果显示广东省生吃水产品中普遍存在常见病原性生物的污染，是食用后引起腹泻等肠胃不适症状以及相关疾病发病率增加的重要原因，应该加强食品卫生管理和食用安全性研究。     

New tools to assess toxicity, bioaccessibility and uptake of chemical contaminants in meat and seafood

Meat and seafood chemical contaminants can be very dangerous for human health. The way in which food chemical contaminants are currently controlled is not optimal as the food cooking, processing and eating habits are generally not considered by authorities. The current review discusses the available information related to the toxicity of the most relevant meat and seafood chemical contaminants, their bioaccessibility after cooking or processing, and the implications for human health. In addition, the current in vivo toxicity and alternative tests carried out for testing the effects of food chemical contaminants are illustrated, as well as new detection tools. The use of non-carcinogenic functional cell models, of alternative animal models like zebrafish embryos, and a toxicogenomic approach seem to be the most promising strategy for the toxicity assessment of food chemical contaminants.