Prevalence and serotyping of Listeria monocytogenes in Portuguese live bivalve molluscs sampled in various steps along the sanitary control process

The prevalence and contamination levels of Listeria monocytogenes were investigated in live bivalve molluscs for human consumption, collected in various steps of the commercial and sanitary circuits in the North of Portugal. Samples of different species were collected per lot before and after depuration treatment in two depuration units and further, when placed in retail markets. Listeria monocytogenes was isolated from 4% of the samples although with very low contamination levels (less than 100 MPN g^?1). None of the environmental (non‐depurated) samples were contaminated with the food‐borne pathogen. The positive samples involved depurated and commercialized samples from two lots, one in each circuit. Two serotypes (1/2a and 3b) were detected among the isolated strains. This study demonstrated the possibility of occurrence of L. monocytogenes contamination of live bivalve molluscs along the sanitary control circuit, including at the retail market level. As L. monocytogenes grows well at refrigerated temperatures and in high salt matrixes such as seafood its presence in these products should not be overlooked.     

Incidence,Growth, and Inactivation of Listeria monocytogenes in Cooked and Peeled Cold-Water Shrimp

ABSTRACT

Cooked and peeled cold-water shrimp (Pandalus jordani) naturally contaminated with Listeria monocytogenes were obtained from a processor for a series of studies to determine the level of contamination and growth characteristics of this bacterium in the naturally contaminated product. L. monocytogenes was isolated from every 25-g sample of individually quick frozen (IQF) shrimp that was tested. The level of contamination in each composite sample ranged from 5 to 16 colony forming units (CFU) per 25 g. When individual shrimp taken from the 25-g sample portions were tested separately, samples positive for L. monocytogenes ranged from 1 of 12 to 5 of 15 shrimp tested. The project also evaluated the effectiveness of three methods to inactivate the bacterium: ozone, chlorine dioxide, and steam as possible product reconditioning strategies. Ozone and chlorine dioxide were both found to be ineffective reconditioning treatments for shrimp naturally contaminated with L. monocytogenes. Experiments with steam conducted at the laboratory and later at the shrimp processing plant verified that shrimp contaminated with L. monocytogenes could be safely reconditioned by steam pasteurization. Steam was used successfully to pasteurize several thousand pounds of contaminated shrimp in the processing plant. When the naturally contaminated product was packaged in either oxygen-permeable or impermeable films and stored at 5°C and 10°C, the product was deemed spoiled by sensory evaluation after 9 days of storage, at which time the L. monocytogenes population were 3 × 10⁴ CFU per g. By comparison, when an isolate (strain 4311) from naturally contaminated shrimp was inoculated onto the pasteurized shrimp at a concentration of 12 cells /25g, the L. monocytogenes population reached 3.0 × 10⁸ per g after 9 days of storage. The pasteurization process used in this study would not be effective in inactivation of Clostridium botulinum. Ready-to-eat-shrimp must therefore be stored below 3°C or frozen.     

Prevalence and Characterization of Coagulase Positive Staphylococci Isolated from Salted Anchovy

A total of 100 salted anchovy samples were used to investigate the prevalence of S. aureus and other coagulase positive Staphylococci (CPS) as well as to determine the methicillin (MR) and antibiotic resistance (AR) profile, the presence of Panton-Valentine leukocidine (PVL) toxin gene (lukS/F-PV), slime factor properties (SFP), and the genotypic relatedness of the isolates. Agar disc diffusion assay (ADDA) and microdilution broth susceptibility test (MDBST) were applied to compare the specificity and sensitivity of the MR detection methods. A total of 41 CPS isolates were detected at the 10² and 10³ CFU/g levels in contrast to S. aureus. The 16S rRNA (genus specific) was detected in all the isolates in contrast to nuc (species-specific) and lukS/F-PV genes. A total of 16/41 isolates were found to be MR by using the three methods. Polymerase chain reaction (PCR) assay was a more sensitive and reliable method for the detection of MR isolates. The antibiotic resistance rates were 75.60, 73.17, 51.21, 31.70, 12.19, and 4.87% to penicillin, ampicillin, tetracycline, erythromycin, ciprofloxacin, and clindamycin, respectively. All the isolates were sensitive to gentamicin and vancomycin. The SFP were determined in all the isolates by using Congo Red agar, and 20 different genotypes were determined by using randomly amplified polymorphic DNA (RAPD)-PCR assay.     

Effect of Organic Acids on Physical-Mechanical and Antifungicidal Properties of Anchovy Protein Films

The use of active biodegradable films from renewable sources like anchovy, with the incorporation of organic acids, such as sorbic acid (SA) and benzoic acid (BA), is an alternative to decrease environmental pollution and minimize the development of fungi in foods. This study investigated the respective influences of SA and BA at 0, 0.50, 0.75, and 1.50% on the growth of Aspergillus flavus and Rhizopus oryzae and on Argentine anchovy protein film properties (L*, a*, b*, tensile strength, elongation at break, solubility in water, water vapor permeability, and Fourier transform infrared spectroscopy (FTIR)). Results showed that the incorporation of SA or BA at different levels significantly increased the water vapor permeability, water solubility, and elongation at break of the films, but decreased the tensile strength and L* parameter (p < 0.05). FTIR evidenced an interaction between the BA and Argentine protein isolate. Also, the influences of the SA or BA concentration on antifungal activity was observed. The greatest effectiveness was obtained for films containing SA against R. oryzae compared to BA film treatments. In conclusion, the Argentine anchovy protein films could retain their antifungal property by incorporation of these organic acids.     

Listeria monocytogenes is inhibited on fillets of cold‐smoked sunshine bass,Morone chrysops × Morone saxatilis,with an edible corn zein‐based coating incorporated with lemongrass essential oil or nisin

Ready‐to‐eat (RTE) foods have been identified as a high‐risk food group because of the number of outbreaks caused by food‐borne pathogens isolated from these products. As these items receive no further processing or heat treatment prior to consumption, bacterial pathogens such as Salmonella, Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes present a serious threat to consumer safety. Of particular concern, L. monocytogenes is resistant to various food storage techniques, including reduced or modified atmosphere packaging, refrigerated storage, and increased salt concentration. Cold‐smoked fishery products have been implicated in a number of listeriosis cases, where it is estimated that between 6 and 36% of cold‐smoked fish is contaminated with L. monocytogenes. Edible coatings incorporated with natural antimicrobials have been suggested to control pathogenic and spoilage bacteria on a variety of meat products. In this study, edible zein‐based coatings incorporated with nisin and lemongrass essential oil (LG) (8%) were evaluated for antibacterial action against L. monocytogenes and spoilage organisms on cold‐smoked, cultured hybrid striped bass, Morone saxatilis × Morone chrysops, under polyvinyl chlorine (PVC) and vacuum packaging for 14 days (PVC) and 42 days (vacuum packaging), respectively, at 4°C. In this study, corn zein‐based edible coatings were found to be an effective carrier for nisin and LG. Nisin‐treated samples were most effective against L. monocytogenes in both PVC and vacuum‐packaged fillets, with a total reduction of 3.5 log and 3.7 log, respectively over the length of the storage time. LG‐treated samples reduced L. monocytogenes cell counts by 2.5 log in PVC and 1.7 log in vacuum‐packaged samples. Only LG‐treated samples packaged in PVC were found to inhibit the growth of spoilage organisms. Because nisin‐ and LG‐treated fillets reduced L. monocytogenes, they may be useful methods to improve food safety in smoked seafood.     

Quality Changes in Marinated Anchovy (Engraulis encrasicolus) Sauced with Olive Oil-Lemon Juice Emulsions

ABSTRACT

This study describes the potential use of olive oil-lemon juice emulsions containing different levels of lemon juice (0, 25, 35, and 50% v/v) as a flavoring, preservative, and antioxidant agent in marinated anchovy. The phenolic content and antioxidant activity of lemon juice, olive oil, and olive oil-lemon juice sauces as well as chemical, oxidative, and sensorial changes of marinade samples were analyzed. Increasing the level of lemon juice in sauces retarded the chemical and oxidative changes of marinated anchovy. However, increasing the level of lemon juice more than 35% in sauces led to a decrease in sensorial acceptability. Chemical and oxidative qualities of all the sauced samples were in the limit of deterioration during 100 days of storage. Although marinated anchovy samples treated with sauce containing 50% lemon juice had a significant inhibitor effect on chemical and oxidative deterioration, samples treated with sauce containing 35% lemon juice received the highest overall acceptability score.     

Thermal Death Time of Listeria monocytogenes Cells in Artificially Contaminated Greenshell Mussels (Perna canaliculus)

ABSTRACT

The production of Greenshell mussel (Perna canaliculus) products involves a heating process, known as a blanching or conditioning step, which facilitates opening and inhibits enzymes which can reduce the shelf-life of mussel products.

To determine whether this initial heating process was also listeri-cidal, thermal death times were determined for seven strains of Listeria monocytogenes in raw Greenshell mussels. D-values at 58, 59, 60,61 and 62^CC were calculated to be 17.19, 11.32, 7.46, 4.91, and 3.24 minutes respectively, giving a z value of 5.52°C. This information combined with information on the core temperatures of mussels during processing will enable processors to implement processing regimes to ensure the elimination of any L. monocytogenes cells associated with the raw product. Such a processing regime has the additional benefit of reducing the carriage of pathogens into the processing facility.     

Quality Changes in Gamma Irradiated Marinades of Anchovy (Engraulis anchoita) During Refrigerated Storage

The effect of gamma irradiation (0, 1.8, and 3.3 kGy) on the microbiological, chemical, and color characteristics of marinated (7% acetic acid and 10% NaCl) and vacuum-packed anchovy fillets was analyzed during 20 months of refrigerated storage (4 ± 1°C). Acidity, pH, water activity (a_w), total volatile basic nitrogen (TVBN), lipid oxidation, and color parameters were determined. Mesophilic and psychrotrophic bacteria, sulphite-reducing clostridia, total and fecal coliforms, Staphylococcus spp., yeasts, and molds were investigated. Gamma irradiation reduced the initial mesophilic bacterial counts and inhibited mesophils growth during 20 months. As a result, the production of TVBN during storage was lower in irradiated samples than in control. Also, lipid oxidation was lower in irradiated samples than in nonirradiated. The color of anchovy fillets was not affected by the irradiation treatment. Even if nonirradiated anchovy fillets presented a high stability in comparison with the traditional product (in flasks with vegetable oil and spices), gamma irradiation improved the microbiological and chemical quality of anchovy fillet marinades without inducing changes on its characteristic color for 20 months.     

Nutritional Characteristics of a Peruvian Anchovy (Engraulis ringens) Protein Concentrate

ABSTRACT

The aim of the present study is to present a new anchovy protein concentrate and compare it to other similar fishery products. A 76.4% protein concentrate was prepared from fresh Peruvian anchoveta (Engraulis ringens) (APC) by means of acid treatment with 1% citric acid, solubilization with 1% sodium chloride, and isoelectric precipitation processes. Remaining fat was 5.2%, and 0.51 A_w indicates good stability. When APC was rehydrated for 10 minutes, it had a 3.5 initial weight increase. It provided 17.8% protein of very high nutritional quality, with only histidine content deficient for children 1–2 years old. Fat content in APC offered 33.4% EPA+DHA. No changes in the essential/non-essential amino acids (EAA/non-EAA), polyunsaturated/saturated fatty acids (PUFA/SFA), or w6/w3 ratios were observed as a result of the process in the raw material. Because of the high sodium content, a 25 g portion is recommended to fulfill nutritional FAO recommendations. APC can be considered as an important source of calcium (30% adults and 35% children requirements), phosphorous (30% adults and 52% children requirements), and iron content (approximately 20% adults and children needs). APC is an important high-quality protein, providing w-3 fatty acids, calcium, phosphorous, and iron.     

Bacterial Contribution to Salted Anchovy (Engraulis anchoita Hubbs & Marinni, 1935) Ripening Process

The bacterial populations of the salted anchovy ripening process and its potential role on the development of the typical sensory characteristics of this product were studied. Salted anchovy samples were taken during the ripening process and submitted to microbiological analyses in order to monitor the evolution of bacterial groups. According to the results obtained, the ripening process was dominated by moderate halophilic bacteria. Moreover, many of the isolated strains showed proteolytic, lipolytic, and trimethylamine oxide (TMAO) reductase activities. These activities contribute to the development of the typical flavor of this product and to the increase of total volatile bases observed during ripening.     

Research Issues in Inactivation of Listeria monocytogenes Associated with New Zealand Greenshell Mussel Meat (Perna canaliculus) Using High-Pressure Processing

ABSTRACT

New Zealand Greenshell mussels are currently shucked by heat processing, and this can be used as a listericidal step. Shucking by high pressure processing (HPP) has potential benefits in product quality and increased yield, but processors need to understand the effects of this technology on the safety of their product with respect to Listeria monocytogenes. L. monocytogenes was mixed with minced mussel meat, and 2 g samples (in foil pouches) were subjected to HPP at various pressures, times, and temperatures. Of 10 tested strains of L. monocytogenes, the most resistant to HPP at 400 MPa (Food Science Australia strain 2655 isolated from Australian processed meat) was selected for subsequent work. This strain showed two-phase inactivation kinetics in response to time at 400 MPa. Approximately 5 log₁₀ cells/g were rapidly inactivated in a log-linear fashion with time while the remaining cells were inactivated at a slower rate. There was also some evidence of a shoulder in the inactivation curves. In the temperature range tested (10°C–40°C), the log-linear inactivation rates showed linear increases with increasing processing temperature at 400 MPa with a z value of 29.1 min.     

Influence of Chitosan Nanocomposite and Rosemary (Rosmarinus officinalis L.) Extract Coating on Quality of Huso huso Fillet Inoculated with Listeria monocytogenes During Refrigerated Storage

This study aimed to evaluate the antilisterial and antioxidative effects of chitosan nanocomposites and rosemary extract coating on the fillet of Huso huso inoculated with Listeria monocytogenes during refrigerated storage (4°C). Fish fillets were subjected as control (without coating), 0.5% rosemary extract (RE), 1% chitosan (CS), and combination of chitosan and rosemary extract as chitosan nanocomposite (CS/RE). Then, samples were inoculated with L. monocytogenes. Subsequently, the chemical parameters (total volatile basic nitrogen (TVB-N), peroxide value (PV), pH, and thiobarbituric acid (TBA)) and antilisterial effects of coatings were monitored during 16 days of storage at 4°C. According to the results, CS/RE demonstrated a significant (p < 0.05) ability to inhibit the growth of L. monocytogenes from 4.14 log cfu/g to 2.23 log cfu/g at the end of the storage period, followed by CT and RET treatments, respectively, compared to the control. Even though samples coated with CS/RE had the lowest pH and TVB-N values (p < 0.05), this coating was not able to pause the protein denaturation after 8 days of storage (p > 0.05) compared to the other treatments. On the other hand, CS/RE coating retarded lipid oxidation by decreasing PV and TBA production in the samples compared to the control up to the end of refrigerated storage (p < 0.05).     

An Intermediate Moisture Product from Mackerel (Rastrelliger kanagurta) Using Salt Curing,Fermentation, and Drying

Intermediate moisture products were prepared from mackerel (Rastrelliger kanagurta) using salt curing in 20% or saturated brine for 20 hr and fermentation with an inoculum level of l0⁸ cells/mL of Pediococcus acidilactici followed by sun drying (28- 33C) for 23 hr or drying in an electric oven (45-50C) for 14 hr to an 18% moisture level. The chemical and microbiological studies correlated with the organoleptic results, suggesting a shelf-life of 4 months for salted and dried products and 7 months for salted, fermented, and dried products.     

Biopreservative effect of pediocin ACCEL on refrigerated seafood

To investigate the biopreservative effectiveness of pediocin ACCEL on refrigerated seafoods, fresh fish fillets were immersed in various concentrations of pediocin ACCEL and then stored at either 4° or 0°C. Samples treated with nisin were used as a positive control. The aerobic plate counts (APC) of samples with bacteriocins were <2.0 log₁₀cfu/g (log cfu/g) after 2 days storage at 0°C, except that with 1500 IU/mL of pediocin ACCEL. The APC of samples with nisin were >2.0 log cfu/g after 2 days storage, while those with pediocin ACCEL occurred after 1 day storage at 4°C. In refrigerated seafoods, pediocin ACCEL and nisin suppressed the growth of inoculated Listeria monocytogenes during 2- and 1-week storage at 4°C, respectively. Compared with nisin, the pediocin ACCEL was considered to be more effective on the suppression of L. monocytogenes growth in refrigerated seafoods during 2-week storage at 4°C.     

Effect of Trisodium Phosphate or Water Dip on the Survival of Salmonella and Listeria monocytogenes Inoculated Catfish Before and After Freezing

Salmonella and Listeria monocytogenes (LM) are occasional contaminants on raw fish. Catfish fillets were artificially contaminated with LM and Salmonella, dipped in a 1.5 % (30 min) trisodium phosphate solution (TSP), and cryogenically frozen. After 3 months frozen storage, Salmonella (2 log), but not LM, was inactivated on the fillets treated with the dip. This indicates that TSP dip followed by cryogenic freezing can be used to control Salmonella, but not LM, on catfish fillets.     

Possible In-Plant Sources of Contamination of Lobster and Snow Crab by Listeria

Abstract

The present study was undertaken to determine potential sources of Listeria monocytogenes contamination through statistical analysis of L. monocytogenes results from lobster and snow crab samples and Listeria spp. results from environmental swabs. The objectives were (1) to compare crustacean analysis records for relationships between presence/absence of L. monocytogenes and crustacean species, year, total aerobes, staphylococci and fecal coli-form; and (2) to identify sites prone to Listeria spp. contamination. There were no significant differences in L. monocytogenes incidence between lobster and snow crab nor between five consecutive years of survey. Total aerobes and staphylococci counts were significantly associated with incidence of L. monocytogenes; fecal coliform counts were not. On surfaces in contact with the processed product, Listeria spp. were mostly found within crab crushers, drum meat separators, shucking machines, on aprons, gloves, work tables and knives. Higher incidence of Listeria spp. were found on surfaces not in direct contact with the processed product such as truck boxes, foot stools, floors, drains and employee boots and pants.     

Protective Effect of a Non-Bacteriocinogenic Lactococcus piscium CNCM I-4031 Strain Against Listeria monocytogenes in Sterilized Tropical Cooked Peeled Shrimp

The protective activity of a non-bacteriocinogenic Lactococcus piscium CNCM I-4031 strain against Listeria monocytogenes was investigated in tropical cooked peeled shrimp stored at 8°C in modified atmosphere packaging (50% N₂–50% CO₂). When inoculated alone (L. piscium 10⁷ CFU g^?1 and L. monocytogenes 10⁴ CFU g^?1), protective culture and target strain grew very well on shrimp reaching a maximum cell number of 10⁹ CFU g^?1 after 7 and 14 days, respectively. In the presence of L. piscium, growth of L. monocytogenes was totally prevented after 3 days of storage. The count was 3.4 log CFU g^?1 lower than in the control after 10 days and until the end of storage (31 days). Using the Seafood Spoilage and Safety Predictor Software (http://sssp.dtuaqua.dk), it was shown that pH decrease from 6.58 to 5.94 and lactic acid concentration of 89.65 mM measured in the co-inoculated batch did not fully explain the inhibition observed.     

Supplementation with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa) in low fish meal diets for the white shrimp,Litopenaeus vannamei

This work evaluated the performance of Litopenaeus vannamei to low fish meal diets supplemented with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa). A basal diet with 150.0 g kg^?1 of anchovy fish meal was designed. Two positive control diets were formulated to reduce fish meal at 50% and 100% with 1.0 and 2.0 g kg^?1 of MERA? Met_Ca (calcium salt with 84% HMTBa activity), respectively. Two nearly equivalent diets acted as negative controls, without HMTBa supplementation. A total of 50 clear‐water tanks of 500 L were stocked with 2.22 ± 0.19 g shrimp under 70 animals m^?2. Shrimp survival (92.3 ± 5.1% and 81.4 ± 8.0%), yield (808 ± 12 and 946 ± 17 g m^?2) and FCR (2.17 ± 0.19 and 3.12 ± 0.37) showed no differences among diets after 72 or 96 days, respectively. A significantly higher shrimp body weight and weekly growth were observed for those fed with the basal diet or diets supplemented with HMTBa compared with non‐supplemented ones. This study has shown that L. vannamei growth, body weight, survival, yield and FCR were supported by HMTBa supplementation when 150.0 g kg^?1 of fish meal was replaced by soybean meal and other ingredients, at 50% and 100%.     

Proximate Composition and Nutritional Profile of the Black Sea Anchovy (Engraulis encrasicholus) Whole Fish,Fillets, and By-Products

ABSTRACT

In this study, the proximate composition and nutritional profile of the Black Sea anchovy and its by-products were investigated. The total yield of by-products from anchovy was about 32% of the whole fish based on wet weight. The protein and fat contents of anchovy by-products were 13.39 and 10.02% for head, 16.47 and 15.50% for frame, and 12.05 and 23.90% for viscera, respectively. Significant differences were detected among the pH and color properties of anchovy whole fish, fillet, and by-products. Profiles of amino acids, fatty acids, and minerals of anchovy by-products showed they are rich sources of lysine (6–7% of total amino acids), leucine (5–6% of total amino acids), and a number of essential amino acid, polyunsaturated fatty acids (about 32–40% of total fatty acids), n-3 fatty acids (about 27–34% of total fatty acids), eicosapentaenoic acid + docosahexaenoic acid (about 26–32% of total fatty acids), and various minerals (P, Fe, Ni, Ca, Mn, Na, and Zn). These results revealed that anchovy by-products can be utilized for the production of value-added products such as protein powder, protein hydrolyzates, fish oils, and mineral supplements.     

Blue mussel meal as feed attractant in rapeseed protein‐based diets for turbot (Psetta maxima L.)

Antinutritional factors in rapeseed products have been identified to reduce feed palatability and growth performance of turbot. Therefore, we evaluated the potential of blue mussel (Mytilus edulis L.) meal as feed attractant in rapeseed protein‐based diets for turbot. Triplicate fish groups received isonitrogenous and isocaloric diets with fish meal (FM) protein replacements of 50% or 75% by rapeseed protein concentrate (RPC 50, RPC 75). These diets were supplemented with 0%, 2%, 4% or 8% of blue mussel meal. In contrast with RPC 50 diets, fish fed with RPC 75/0 showed significantly reduced daily feed intake (DFI) and specific growth rate (SGR). With increasing mussel meal inclusion, RPC 75 diets resulted in increased DFI and SGR, suggesting mussel meal as attractant in rapeseed protein‐based diets for turbot. Feed conversion was unaffected by any treatment. Protein productive value and apparent digestibility coefficients were unaffected by either RPC or mussel meal inclusion. With regard to the whole body composition, no differences in crude protein, crude lipid and ash content could be determined. Haematological characteristics were unaffected among the treatments indicating good nourished and unstressed fish. In conclusion, we demonstrated that the utilization of blue mussel meal improved the palatability of rapeseed protein‐based diets for turbot.