Identification of arginine kinase as an allergen of brown crab, Callinectes bellicosus, and in silico analysis of IgE-binding epitopes.

In recent years there has been an increase in the prevalence of allergic reactions to contact with/or consumption of crustaceans by immune responses mediated by IgE antibodies. Arginine kinase (AK) is considered one of the main allergens present in marine invertebrates. Currently, the allergenic potential of the brown crab (Callinectes bellicosus), which is a crustacean of great economic importance, has not been studied. Therefore, the aim of this work was to identify C. bellicosus AK as an allergen and to predict IgE-binding epitopes through immunobioinformatic analysis. AK was purified by precipitation with ammonium sulfate and ion- exchange chromatography. AK allergenicity was evaluated by IgE reactivity against sera from crustacean-allergic and non-allergic patients in both native and denaturing conditions. Additionally, a homology model was built based on the deduced amino acid sequence. A single band (~40 kDa) was found in SDS-PAGE, which was identified as an AK by mass spectrometry. AK showed immunoreactivity against crab-allergenic sera in both native and denaturing conditions with 70% and 80% positive reactions, respectively. Additionally, a 1073 bp ORF was obtained which codes for a deduced sequence of 357 amino acids corresponding to AK with > 90% identity with other AKs. Structural homology model of AK showed two main domains with conserved / folding of phospho-guanidine kinases. BediPred and Discotope were used for epitope prediction analysis, which suggests eight possible linear epitopes and seven conformational epitopes, respectively; and shows to be similar to other crustaceans AKs. C. bellicosus AK was identified as an allergenic protein by IgE reactivity and immunobioinformatic analysis indicates that both linear and conformational epitopes could be located in the surface of C. bellicosus AK structure.

Shell water-soluble matrix protein from oyster shells promoted proliferation, differentiation and mineralization of osteoblasts in vitro and vivo.

Matrix protein is secreted by the membrane of bivalve shellfish to and used to regulate shell biomineralization. In this study, we extracted water-soluble matrix protein (WSMP) from oyster shells to investigate its effects on osteogenic differentiation and mineralization of MC3T3-E1 cells and osteoporosis rats. Our results suggested that WSMP was an acidic glycoprotein by amino acid analysis and secondary structure analysis. In vitro, WSMP could promote osteoblastic proliferation. Moreover, alkaline phosphatase (ALP) and osteocalcin (OCN) were increased, mineralized nodules were increased, and BMP-2 expression was up-regulated. Additionally, in vivo, tartrate-resistant acid phosphatase (TRAP) and Bone alkaline phosphatase (BALP) expressions in the medium-dose and high-dose groups were significantly decreased compared with the model group, while OCN expression was significantly increased. Bone mineral density (BMD) and bone mineral content (BMC) of bone recovered significantly. In summary, WSMP can promote the proliferation, differentiation and mineralization of osteoblasts in vitro and in vivo.

IgE epitope analysis of sarcoplasmic-calcium-binding protein, a heat-resistant allergen in Crassostrea angulata.

Sarcoplasmic-calcium-binding protein (SCP) has been investigated as a novel allergen in Crassostrea angulata. Nevertheless, knowledge of its effector-cell-based allergic relevance and epitopes is limited. In this study, the heat-resistant allergen SCP was able to induce significant upregulation of CD63 and CD203c (p < 0.05), which showed obvious allergenicity in a basophil activation test. Furthermore, immunoinformatic tools, a one-bead-one-compound peptide library, and phage display technology were combined to analyze the allergenic epitopes of SCP. Five linear epitopes named L-SCP-1 (AA22-33), L-SCP-2 (AA64-75), L-SCP-3 (AA80-90), L-SCP-4 (AA107-116), and L-SCP-5 (AA144-159) were verified using serological tests. Additionally, two conformational epitopes (C-SCP-1 and C-SCP-2) were determined, and C-SCP-1 was located at one of the calcium-binding sites (AA106-117). Moreover, SCP showed weaker typical α-helical features and higher hydrophobicity after Ca2+ depletion, which reduced its IgE-binding capacity. Overall, these epitope data could enhance our understanding of oyster allergens, which could be used to develop hypoallergenic shellfish products.

Fish and Shellfish-Derived Anti-Inflammatory Protein Products: Properties and Mechanisms.

The interest in utilizing food-derived compounds therapeutically has been rising. With the growing prevalence of systematic chronic inflammation (SCI), efforts to find treatments that do not result in the side effects of current anti-inflammatory drugs are underway. Bioactive peptides (BAPs) are a particularly promising class of compounds for the treatment of SCI, and the abundance of high-quality seafood processing byproducts (SPB) makes it a favorable material to derive anti-inflammatory BAPs. Recent research into the structural properties of anti-inflammatory BAPs has found a few key tendencies including they tend to be short and of low molecular weight (LMW), have an overall positive charge, contain hydrophobic amino acids (AAs), and be rich in radical scavenging AAs. SPB-derived anti-inflammatory BAPs have been observed to work via inhibition of the NF-κB and MAPK pathways by disrupting the phosphorylation of IκBα and one or more kinases (ERK, JNK, and p38), respectively. Radical scavenging capacity has also been shown to play a significant role in the efficacy of SPB-derived anti-inflammatory BAPs. To determine if SPB-derived BAPs can serve as an effective treatment for SCI it will be important to understand their properties and mechanisms of action, and this review highlights such findings in recent research.

Environmental salinity and dietary lipid nutrition strategy: Effects on flesh quality of the marine euryhaline crab Scylla paramamosain.

The quality of crustaceans' flesh has direct impact on consumers' purchase choices, with water environment and dietary nutrition being effective ways to regulate flesh quality. The aim of present study was to investigate the impacts of water salinity (low, 4 and medium, 23) and dietary lipid source (fish oil and soybean oil) on nutritional values, texture, taste and odor of flesh of mud crab. While water salinity had no significant influence on nutritional values of crab flesh, crabs fed soybean oil displayed significantly lower contents of amino acids and n-3 PUFAs in muscle. However, crabs reared at low salinity showed reduced flesh hardness, chewiness and gumminess likely related to altered myofiber structure, that impacted muscle texture. Furthermore, low salinity and dietary soybean oil weakened umami taste and aroma characteristics of crab flesh associated with decreased contents of free amino acids, flavor nucleotides, inorganic ions and odor active compounds in flesh.

Insights into the similarities and differences of whiteleg shrimp pre-soaked with sodium tripolyphosphate and sodium trimetaphosphate during frozen storage.

In this study, similarities and differences of sodium tripolyphosphate (STPP) and sodium trimetaphosphate (STMP) pre-soaking on the stability of muscle proteins in shrimp were investigated during 12 weeks of frozen storage (-30 °C). The physicochemical analysis indicated significant improvements in the WHC, springiness, chewiness, and thermal stability of STPP and STMP pre-soaked samples when compared to the control. Interestingly, STMP pre-soaking showed better cryoprotective effects than the STPP treatment when the storage period reached the end of the 12 weeks. Furthermore, the label-free based proteomics results indicated that 62 upregulated differentially abundant proteins (DAPs) were detected in STMP when compared to STPP. These identified DAPs specifically included 40S ribosomal proteins, actin-related proteins, heat shock proteins, myosin heavy chain, and tubulin beta chain. Additionally, the gene ontology (GO) and eukaryotic clusters of orthologous group (KOG) analyses verified that the incorporation of STMP molecules enhanced the resistance of cytoskeleton proteins to cold-temperature stress.

Meat texture, muscle histochemistry and protein composition of Eriocheir sinensis with different size traits.

The main aim of the present work was to investigate the effect of the inherent differences in the raw muscle on the textural quality of the cooked meat from different sized Eriocheir sinensis. The content of entrapped water was 73.8-77.7 g/100 g in raw muscle. The density and diameter of muscle thick microfilaments ranged between 137 and 158/µm2 and 20.9-27.0 nm. These results demonstrated that the raw muscle from the tender group had a smaller density of larger diameter thick microfilaments and more entrapped water than other samples, which could explain the high tenderness of the cooked meat (P < 0.05). The potential structural proteins that indicated tenderness include myosin regulatory light chain 2, ancient ubiquitous protein 1, tubulin α-2 chain, and ß-catenin, was determined using liquid chromatography-tandem mass spectrometry. The inherent attributes of raw muscle could explain the textural differences of the cooked meat from Eriocheir sinensis.

Restriction site-associated DNA sequencing (RAD-seq) analysis in Pacific oyster Crassostrea gigas based on observation of individual sex changes.

The diverse modes of sexual reproduction in Bivalvia make it an excellent clade to understand the evolution of sex and sex determination. The cosmopolitan Pacific oyster Crassostrea gigas is an ideal model for bivalve sex determination studies because of its complicated sexuality, including dioecy, sex change and rare hermaphroditism. A major barrier to C. gigas sex determination study has been the lack of information on the type of sex determination. To identify its sex-determining system, sex observation by following the same individual in two consecutive years was conducted on 760 oysters from distinct populations. Stable sexuality and sex reversal in both directions were observed, which provides a case against the protandry of C. gigas. Restriction site-associated DNA sequencing (RAD-seq) based on 26 samples with unchanged and converted sexualities was carried out for identifying sex-linked marker. One SNP Cgsl-40 was proved to be sex-related, but sex-biased heterozygosity varied between populations for RAD-seq and validation, showing no evidence for sex chromosomes or single-locus models for C. gigas primary sex determination. Information obtained in our study provides novel insight into sex determination mechanism in C. gigas.

A novel blue crab chitosan/protein composite hydrogel enriched with carotenoids endowed with distinguished wound healing capability: In vitro characterization and in vivo assessment.

This work aimed to the development of chitosan and protein isolate composite hydrogels, for carotenoids-controlled delivery and wound healing. By increasing the concentration of the protein isolate, chitosan hydrogels were more elastic at a protein isolate concentration not exceeding 15% (w/w). Chitosan-protein isolate composite hydrogels revealed low cytotoxicity towards MG-63 osteosarcoma cells. Thanks to its appropriate structural, swelling and mechanical resistance properties, chitosan hydrogel (3%; w/v), reinforced with 15% (w/w) of protein isolate, was selected for the carotenoids in vitro release study. Release profiles, show delivery patterns, where carotenoids were more barely released at a pH 7.4 medium (p < .05), compared to more acidic microenvironments (pH 4.0 and pH 2.0). Thus, developed hydrogels could be applied as pH-sensitive intelligent carriers, for drugs-controlled release, with interesting antioxidant abilities. The in vivo healing potential of hydrogels in rats' models was further studied. Topical application of hydrogel-based patches allowed the acceleration of wound healing and the complete healing, for composite hydrogel enriched with carotenoids.

Partially degraded chitosan-based flocculation to achieve effective deodorization of oyster (Crassostrea gigas) hydrolysates.

Aquatic protein hydrolysates are usually associated with unpleasant odors and high fat content, which seriously restricts their industrial utilization. In this study, chitosans with different molecular weights produced by hydrogen peroxide degradation were applied to establish a flocculation method, using for the deodorization and defatting of oyster (Crassostrea gigas) hydrolysates. GC-MS analysis showed that the method markedly decreased the content of the fishy odor constituents. Up to 92 % fat and part of the heavy metals were effectively removed. Protein recovery percentage and solid recovery percentage were 83.43 ± 0.35 % and 76.36 ± 0.52 %, respectively, at the optimum dose (150 mg/L) of chitosan (83 % of deacetylation degree, 77 kDa). Thus, chitosan flocculation-coupled centrifugation (5000g, 1 min) can effectively solve the current drawbacks of engineering disc centrifuges and can be industrially used for defatting and deodorization during aquatic food processing.

Insight into the allergenicity of shrimp tropomyosin glycated by functional oligosaccharides containing advanced glycation end products.

Tropomyosin (TM) is the main allergen of shrimp. Glycation reportedly reduced the allergenicity of TM, and the allergenicity reduction was heavily dependent upon the sources of saccharides. In this work we investigated, how glycation of tropomyosin by functional oligosaccharides affected the allergenicity. Compared to TM, the TM glycated by galacto-oligosaccharide (TM-GOS), mannan-oligosaccharide (TM-MOS) and maltopentaose (TM-MPS) had lower allergenicity and induced weaker mouse allergy responses. While the TM glycated by fructo-oligosaccharide (TM-FOS) had stronger allergenicity and induced severe mouse allergy symptoms, due to the generation of neoallergns that belonged to advanced glycation end products (e.g. CML). Therefore, GOS, MOS and MPS could be applied to desensitize shrimp TM-induced food allergy through glycation, while FOS was not suitable to reduce TM allergenicity. Glycation of TM by GOS, MOS and MPS, especially for MPS, significantly reduced allergenicity and alleviated allergy symptoms, which could be potentially explored for immunotherapy for shrimp-allergic patients.

Evaluation of physicochemical and functional properties of spray-dried protein hydrolysate from non-penaeid shrimp (Acetes indicus).

BACKGROUND: Protein hydrolysate powder was prepared from non-penaeid shrimp (Acetes indicus) by enzymatic hydrolysis using Alcalase enzyme. Extraction conditions such as pH (6.5, 7.5 and 8.5), enzyme to substrate ratio (1.0, 1.5 and 2.0) and temperature (40, 50 and 60 °C) were optimized against the degree of hydrolysis using response surface methodology. RESULTS: Protein hydrolysate comprised of 740 g kg-1 protein, 150 g kg-1 ash and 90 g kg-1 fat contents. The amino acid score showed superior attributes with 56% essential amino acids. Furthermore, the functional properties of spray-dried protein hydrolysates were evaluated. Protein solubility was found to be the 90.20% at pH 2 and 96.92% at pH 12. Emulsifying properties were found to vary with the concentration of protein hydrolysates and the highest emulsifying capacity (26.67%) and emulsion stability (23.33%) were found at a concentration of 20 mg mL-1 . The highest and the lowest foaming capacity were observed at pH 6 and pH 10 with a concentration of 20 mg mL-1 . The water holding capacity of protein hydrolysate was found to increase with concentration, with a value of 5.4 mL g-1 at a concentration of 20 mg mL-1 . CONCLUSION: The results of the present study indicate that the use of A. indicus for the production of protein hydrolysate has good functional properties and nutritional value, rendering it suitable for broad industrial food applications. © 2019 Society of Chemical Industry.

Comparative iTRAQ-based quantitative proteomic analysis of the Chinese grass shrimp (Palaemonetes sinensis) infected with the isopod parasite Tachaea chinensis.

BACKGROUND: Although parasitic isopods can negatively affect the reproduction and ingestion of several commercially important crustaceans, little is known regarding the mechanisms that underlie these effects. METHODS: In the present study, the iTRAQ (isobaric tags for relative and absolute quantification) approach was applied to identify differentially expressed proteins in the Chinese grass shrimp Palaemonetes sinensis infected with the parasitic isopod Tachaea chinensis. RESULTS: On the basis of our analysis, we identified 1262 proteins from a total of 4292 peptides. There was a significant difference in the expression of 182 proteins between the control and infected groups, among which 69 were upregulated and 113 were downregulated after T. chinensis infection. The differentially expressed proteins revealed that parasitism may inhibit the immune response, thereby increasing host vulnerability to additional lethal infection. Furthermore, T. chinensis may secrete anticoagulants to inhibit hemolymph clotting. Moreover, the isopod parasite placed a heavy metabolic burden on the host, particularly with respect to glucose metabolism. CONCLUSIONS: Our study is the first to use the iTRAQ-based proteomic approach to analyze the effects of an isopod parasite on its host. The results we obtained using this approach will make a valuable contribution to understanding the molecular mechanisms underlying isopod parasitism on crustaceans.

The Single-molecule long-read sequencing of Scylla paramamosain.

Scylla paramamosain is an important aquaculture crab, which has great economical and nutritional value. To the best of our knowledge, few full-length crab transcriptomes are available. In this study, a library composed of 12 different tissues including gill, hepatopancreas, muscle, cerebral ganglion, eyestalk, thoracic ganglia, intestine, heart, testis, ovary, sperm reservoir, and hemocyte was constructed and sequenced using Pacific Biosciences single-molecule real-time (SMRT) long-read sequencing technology. A total of 284803 full-length non-chimeric reads were obtained, from which 79005 high-quality unique transcripts were obtained after error correction and sequence clustering and redundant. Additionally, a total of 52544 transcripts were annotated against protein database (NCBI nonredundant, Swiss-Prot, KOG, and KEGG database). A total of 23644 long non-coding RNAs (lncRNAs) and 131561 simple sequence repeats (SSRs) were identified. Meanwhile, the isoforms of many genes were also identified in this study. Our study provides a rich set of full-length cDNA sequences for S. paramamosain, which will greatly facilitate S. paramamosain research.

Physicochemical changes of myofibrillar proteins of squid (Argentinus ilex) induced by hydroxyl radical generating system.

The effect of a hydroxyl radical generating system (HRGS), which contained FeCl3, sodium ascorbate, and different concentrations of H2O2, on the physiochemical properties of myofibrillar protein (MP) from squid mantles, has been investigated. The effect of different exposure times to HRGS was also considered. Compared to non-oxidized MP, a significant (p < 0.05) increase in carbonyl content (more than 50% of its original content) and protein solubility, as well as in surface hydrophobicity, was observed in the oxidative MP. With different treatment times, a sharp decrease (p < 0.05) in sulfhydryl content was detected. In addition, hydroxyl radical treatment significantly reduced the MP gel's texture properties, whiteness and water holding capacity, especially at higher concentrations of H2O2. This observation could be attributed to extensive disorderly and less compact structure of MP gels. The results demonstrate the negative effect of HRGS on the structural and functional properties of MP from squid mantles.

A chiral assembly of gold nanoparticle trimer-based biosensors for ultrasensitive detection of the major allergen tropomyosin in shellfish.

A biosensor based on a chiral assembly of polymer of gold nanoparticle (AuNP) trimers was developed for the detection and quantification of the major shellfish allergen tropomyosin (TROP). TROP and anti-TROP monoclonal antibodies (mAb) were immobilized on 20 nm and 30 nm 16-mercaptohexadecanoic acid (16-MHDA) functionalized AuNPs to assemble a trimer, which has a Circular dichroism (CD) signal. The free TROP from samples was quantified as an inhibitor for the formation of the AuNP trimer. The AuNP trimer-based biosensor allowed for the selective determination of TROP in the range of 0.1-15 ng mL-1 with the limit of detection (LOD) of 21 pg mL-1 (S/N = 3) and the limit of quantitation (LOQ) of 70 pg mL-1 (S/N = 10). The use of a AuNP trimer-based biosensor with simple sample preparation functions with specificity and accuracy; this highlights its applicability for the detection of allergens in shellfish products, related products and their production lines. Furthermore, based on the less conserved sequences of TROP in phylogenetically different species, this biosensor is currently being used to identify the adulteration of shellfish products using TROP as biomarker.

High-Pressure Homogenization Pre-Treatment Improved Functional Properties of Oyster Protein Isolate Hydrolysates.

The effects of HPH (high-pressure homogenization) pre-treatment on the functional properties of OPIH (oyster protein isolates hydrolysates) were studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles, solubility, particle size distribution, zeta potential, surface hydrophobicity, emulsifying activity index and microstructure of emulsions were analyzed. Results indicated that HPH pre-treatment increased the accessibility of OPI to trypsin hydrolysis, resulting in decease in particle size, increase in solubility, absolute zeta potential, surface hydrophobicity and emulsifying activity index. In addition, HPH pre-treated OPIH emulsions became more uniform and the particle size of droplets decreased. These results revealed that HPH pre-treatment has the potential to modify the functional properties of OPIH.

Dual Gene Repertoires for Larval and Adult Shells Reveal Molecules Essential for Molluscan Shell Formation.

Molluscan shells, mainly composed of calcium carbonate, also contain organic components such as proteins and polysaccharides. Shell organic matrices construct frameworks of shell structures and regulate crystallization processes during shell formation. To date, a number of shell matrix proteins (SMPs) have been identified, and their functions in shell formation have been studied. However, previous studies focused only on SMPs extracted from adult shells, secreted after metamorphosis. Using proteomic analyses combined with genomic and transcriptomic analyses, we have identified 31 SMPs from larval shells of the pearl oyster, Pinctada fucata, and 111 from the Pacific oyster, Crassostrea gigas. Larval SMPs are almost entirely different from those of adults in both species. RNA-seq data also confirm that gene expression profiles for larval and adult shell formation are nearly completely different. Therefore, bivalves have two repertoires of SMP genes to construct larval and adult shells. Despite considerable differences in larval and adult SMPs, some functional domains are shared by both SMP repertoires. Conserved domains include von Willebrand factor type A (VWA), chitin-binding (CB), carbonic anhydrase (CA), and acidic domains. These conserved domains are thought to play crucial roles in shell formation. Furthermore, a comprehensive survey of animal genomes revealed that the CA and VWA-CB domain-containing protein families expanded in molluscs after their separation from other Lophotrochozoan linages such as the Brachiopoda. After gene expansion, some family members were co-opted for molluscan SMPs that may have triggered to develop mineralized shells from ancestral, nonmineralized chitinous exoskeletons.

Enumeração e identificação de víbrios sacarose-positivos em amostras de lulas frescas comercializadas no município de Niterói-RJ / Enumeration and identification of sucrose-positive vibrios in samples of fresh squids commercialized at Niterói county-RJ

Foi efetuado um estudo sobre a enumeração e a identificação de víbrios sacarose-positivos em lulas frescas obtidas no comércio varejista do município de Niterói-RJ. No experimento, em 50 amostras de lulas, identificadas como pertencentes à espécie Oorytheutis brasiliensis Blainville, 1823, foram isoladas Unidades Formadoras de Colônias (UFC) sacarose-positivas em 28 (56%) amostras. Destas, foram identificados o Vibrio a/ginolyticus e víbrios do grupo NAG (não aglutináveis). A média dos Números Mais Prováveis (NMP) dos víbrios totais (com exceção da amostra com NMP > 2400 bacts./g) foi 101,70 bacts./g. Do total de 29 isolamentos nas 28 amostras, o V alginolyticus do grupo I de Heiberg obteve o maior percentual (62,07%), o V alginolyticus do grupo 111 apenas 10,30% e os víbrios NAG com 27,63%.

From 50 samples of squids of lhe species Dorytheutis brasiliensis Blainville, 1823, Colonies Formed Units (CFUs) sucrose-positives Víbrio alginolyticus and NAG vibrios (non-agglutinable) were isolated. The average of the Most Probabble Number of total vibrios (except the sample with MPN > 2400 bacts./g) was 101,70 bacts./g. From 28 samples, 29 isolations were carried out and lhe V. alginolyticus group I of Heiberg's Classification showed the greatest percentage (62,07%). The V. alginolyticus group 111 showed only 10,30% and NAG vibrios 27,63%.

Enumeração e identificação de Vibrio parahaemolyticus em lulas frescas comercializadas no município de Niterói RJ Brasil / Enumeration and identification of Vibrio parahaemolyticus in fresh squids commercialized at Niterói, Rio de Janeiro State, Brazil

Com o objetivo de verificar as condições higiênico-sanitárias do pescado comercializado no município de Niterói, efetuou-se um estudo sobre a enumeração e identificação de Vibrio parahaemolyticus em lulas frescas. Foram utilizadas 50 amostras de lulas frescas, identificadas como pertencentes à espécie Oorytheutis brasiliensís Blainville, 1823. A identificação e enumeração do V. parahaemolyticus foram baseadas nos métodos descritos pela lnternational Commission on Microbiological Specifications for Foods - ICMSF (1983). A classificação de Heiberg (1936) foi utilizada como teste complementar. O V. parahaemolyticus foi identificado em três (6%) amostras, sendo que, em uma o vibrio se desenvolveu em meio com concentração salina a 10%, na prova de halofilismo. O V. parahaemolyticus apresentou os NMPs (Número Mais Provável) com média de 19,3.bactérias/g. As três amostras de V. parahaemolyticus submetidas à classificação de Heiberg apresentaram perfil compatível com o grupo VIl (Sacarose -, Manose + e Arabinose +). Concluiu-se que, mesmo sob comercialização inadequada, com manipulação e resfriamento precários, as amostras analisadas apresentaram o V. parahaemolyticus com frequências e NMPs baixos.

A study on the enumeration and identification of Vibrio parahaemolyticus in freshsquids in Niterói was undertook aiming at checking the hygienic-sanitary conditions of fish commercialized in that City. Fifty fresh squids (Oorytheutis brasiliensis, Blainville, 1823). Methods described by the lnternational Comission on Microbiological Specification for Foods-ICMSF (1983) were used to identify and enumerate V. parahaemoiyticus and Heiberg's classi-fication was a complementai test. v. parahaemoiyticus was identified in three samples (6%); in one of them it grew in an environment with 10% salt concentration during halophilism probe. Most probable number (MPN) average for V. parahaemoiyticus was 19,3 bacteria per gram. The three samples submitted to Heiberg's classification showed a profile compatible with group VIl (sacarose-, manose+, arabinose-). We concluded that, even under inadequate commercialization, manipulation and refrigeration conditions, V. parahaemolyticus frequencies and MPNs are low.