Aims: Escherichia coli O157:H7 is known to be transmitted via fecal-oral route, where water plays a role in the transmission process. Oysters as bivalves, bio accumulate pathogens from the water through filter feeding and are suspected to play a role as disease transmission vector. In Malaysia, the data on oyster’s microbiological quality are limited. Hence, it was vital to conduct oyster related studies in Malaysia. The main objectives of this study include the enumeration of most probable number (MPN) of fecal coliforms and E. coli and isolation of E. coli from oyster (Crassostrea iredalei) and water sample for the detection of 16S rRNA and HlyA (Hemolysin A) genes of E. coli O157:H7. Methodology and results: A total of 120 oysters and water samples (n=6) were collected from a fisherman village located in southern Malaysia. Total fecal coliforms and E. coli were determined using the MPN procedure. Colonies of E. coli were identified based on Gram staining, biochemical test, and PCR detection for the presence of 16S rRNA and HlyA gene of E. coli O157:H7. The enumeration results showed that the MPN of the fecal coliforms and E. coli found in the collected oyster samples do not meet the standard to be directed for human consumption (0.72 ± 0.19 × 104 MPN/100 g and 0.13 ± 0.03 × 10 4 MPN/100 g, respectively). The PCR assays showed that 16 out of the 104 (15.38%) of E. coli isolated from water and oysters showed the presence of HlyA gene. The phylogenetic tree analysis showed there were genetic relationships between the HlyA gene of the E. coli isolated in this study with the ones isolated from calf and human faeces. Conclusion, significance and impact of study The detection of Shiga toxin producing E. coli O157:H7 (HlyA gene) in cage cultured oysters (C. iredalei) and water from southern Malaysia was first time reported here. In the future, more study can be conducted to study the expression of the HlyA gene and confirm of its identity as E. coli O157:H7 using different target genes such as eaeA (encodes a 94 kD outer membrane protein called intimin) and Stx1 (Shiga toxin, Shigella dysenteriae type 1).
Escherichia coli O157 ; Crassostrea

Crassostrea sikamea (C.sikamea) is an important edible and medicinal seafood in China. In the present study, a compound named flazin was separated and identified from the ethyl acetate extract of C.sikamea (EAECs) for the first time. In addition, the 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra zolium (MTS) assay revealed that EAECs and flazin inhibited the transformation of splenic lymphocytes in vitro. Moreover, flazin (20 μg·mL
Animals ; Carbolines ; Crassostrea ; Furans ; Lymphocytes ; Rats ; Rats, Sprague-Dawley ; Spleen

Defensins are endogenous cationic antimicrobial peptides rich in arginine and cysteine residues. They are important immune factors resisting pathogenic bacteria infection for mollusks. The 43 amino acid residues near the carboxyl terminal for Crassostrea gigas defensin (CgD) form its mature peptide region, responsible for the biological activity of CgD. First, two target genes, CgDH⁺ (with 6×His-tag at 3' end) and CgDH- (without 6×His-tag at 3' end) were separated and amplified by RT-PCR with specific primers from Crassostrea gigas mantle. These two target genes were ligated to the expression vector pPICZαA to construct recombinant expression vectors, pPICZαA-CgDH⁺ and pPICZαA-CgDH-, which were transformed into competent Pichia pastoris X-33 cells by electroporation respectively. The recombinant target proteins, CgDH⁺ and CgDH-, were induced for 72 h with 1% methanol at 29 °C and 250 r/min. The recombinant CgDH⁺ (5.78 kDa) was purified by immobilized metal affinity chromatography (IMAC), and identified by MALDI-TOF-TOF analysis, demonstrating that it was the expected target protein. Based on the concentration of the purified product, the estimated yield of recombinant CgDH⁺ was 2.32 mg/L. Antimicrobial assay showed that the culture medium supernatant containing recombinant CgDH⁺ and recombinant CgDH-, respectively, had activities against Staphylococcus aureus and Pseudomonas aeruginosa, indicating that the existence of 6×His tag in the recombinant proteins do not affect their biological activities.
Animals ; Anti-Bacterial Agents ; Antimicrobial Cationic Peptides ; Crassostrea ; Defensins ; Pichia ; Recombinant Proteins

In order to assess changes in the activity of immunecompetency present in Crassostrea gigas infected with Marteilioides chungmuensis (Protozoa), the total hemocyte counts (THC), hemocyte populations, hemocyte viability, and phagocytosis rate were measured in oysters using flow cytometry. THC were increased significantly in oysters infected with M. chungmuensis relative to the healthy appearing oysters (HAO) (P<0.05). Among the total hemocyte composition, granulocyte levels were significantly increased in infected oysters as compared with HAO (P<0.05). In addition, the hyalinocyte was reduced significantly (P<0.05). The hemocyte viability did not differ between infected oysters and HAO. However, the phagocytosis rate was significantly higher in infected oysters relative to HAO (P<0.05). The measurement of alterations in the activity of immunecompetency in oysters, which was conducted via flow cytometry in this study, might be a useful biomarker of the defense system for evaluating the effects of ovarian parasites of C. gigas.
Animals ; Cell Count ; Cell Survival ; Cercozoa/*immunology/*pathogenicity ; Crassostrea/*immunology ; Flow Cytometry ; Granulocytes/immunology ; Hemocytes/immunology ; Phagocytosis

A survey was performed to know the recent infection status of digenetic trematode metacercariae in clams and oysters from 4 sites in western coastal regions of the Republic of Korea (=Korea). Four species of clams (Mactra veneriformis, Ruditapes philippinarum, Cyclina sinensis, and Saxidomus purpuratus) were collected from Taean-gun, Chungcheongnam-do (Province), Buan-gun (County) and Gochang-gun, Jeollabuk-do, and oysters, Crassostrea gigas, from Shinan-gun, Jeollanam-do were transferred to our laboratory on ice and examined by the artificial digestion method. The metacercariae of Himasthla alincia were detected in 3 species of clams, M. veneriformis, R. philippinarum, and C. sinensis from the 3 surveyed areas. The positive rate and the mean density per clam infected were 98.9% (30.8 metacercariae) in M. veneriformis, 60.0% (5.0) in R. philippinarum, and 96.0% (28.4) in C. sinensis. The positive rate (mean density) of Acanthoparyphium tyosenense metacercariae in M. veneriformis was 50.0% (2.1) from Taean-gun and 70.0% (2.8) from Gochang-gun. The metacercariae of Parvatrema spp. were detected in M. veneriformis and R. philippinarum from Taean-gun and Gochang-gun; the positive rate (mean density) was 63.3% (4,123) and 50.0% (19) in M. veneriformis, and 6.7% (126) and 100% (238) in R. philippinarum from the 2 regions, respectively. The metacercariae of Gymnophalloides seoi were detected in all 30 oysters from Shinan-gun, and their average density per oyster was 646. From the above results, it has been confirmed that more than 3 species of metacercariae are prevalent in clams from the western coastal regions, and G. seoi metacercariae are still prevalent in oysters from Shinan-gun, Jeollanam-do, Korea
Bivalvia* ; Chungcheongnam-do ; Crassostrea ; Digestion ; Ice ; Jeollabuk-do ; Jeollanam-do ; Korea ; Metacercariae* ; Methods ; Ostreidae* ; Prevalence* ; Republic of Korea* ; Trematoda

In intestinal helminth infections, Th2 immune respones are generally associated with mucin secretion for worm expulsion from the host intestine. In particular, IL-4 and IL-13 are the important cytokines related with intestinal mucus production via STAT6 signalling in nematode infections. However, this perspective has never been studied in Gymnophalloides seoi infection. The present study aimed to observe the STAT6 signalling and cytokine responses in C57BL/6 mice, a mouse strain resistant to infection with this trematode. The results showed that worm expulsion occurred actively during days 1-2 post-infection (PI), when goblet cells began to proliferate in the small intestine. The STAT6 gene expression in the mouse spleen became remarkable from day 2 PI. Moreover, G. seoi infection induced a significant increase of IL-13 from day 4 PI in the spleen of infected mice. Our results suggested that goblet cell hyperplasia and worm expulsion in G. seoi-infected mice should be induced by STAT6 signalling, in which IL-13 may be involved as a dominant triggering cytokine.
Animals ; Crassostrea ; Female ; Goblet Cells/*pathology ; Hyperplasia/pathology ; Interleukin-13/*metabolism ; Interleukin-4/*metabolism ; Intestine, Small/immunology ; Metacercariae ; Mice ; Mice, Inbred C57BL ; STAT6 Transcription Factor/*metabolism ; Signal Transduction ; Specific Pathogen-Free Organisms ; Spleen/immunology ; Trematoda/*immunology ; Trichinellosis/*immunology/parasitology

OBJECTIVECrassostrea gigas oyster extract has been reported to have antioxidant, antihypertensive and lipid-lowering properties that may be useful for treating cardiovascular diseases. This study aimed to evaluate the effect of C. gigas oyster extract on cardiovascular function in tissues from healthy rats.

METHODSSingle-cell microelectrode and isolated thoracic aortic organ bath studies were performed on tissues from 8-week-old healthy Wistar rats, using varying concentrations of C. gigas oyster extract. To elucidate a mechanism of action for the oyster's vasoactive properties, concentration response curves were carried out in the presence of a calcium channel inhibitior (verapamil), a nitric oxide synthase inhibitor (N(G)-nitro-L-arginine methyl ester), a potassium channel inhibitor (4-aminopyridine), in addition to the α-adrenoceptor inhibitor prazosin.

RESULTSOyster solution at 7 500 mg/mL inhibited noradrenaline-induced contraction in isolated aortic rings. Cardiac electrophysiology results showed that neither concentration of oyster solution was able to significantly reduce action potential duration at all phases of repolarisation in left ventricular papillary muscles from healthy animals.

CONCLUSIONWhen administered to healthy vascular tissue, C. gigas oyster extract inhibits contraction induced by noradrenaline. This effect is likely to be mediated through α-adrenoceptor inhibition, and to a lesser extent, calcium modulating activity.

Action Potentials ; drug effects ; Adrenergic alpha-Antagonists ; pharmacology ; Animals ; Aorta, Thoracic ; drug effects ; physiology ; Calcium ; metabolism ; Crassostrea ; Heart ; drug effects ; physiology ; Male ; Norepinephrine ; antagonists & inhibitors ; pharmacology ; Rats ; Rats, Wistar ; Vasoconstriction ; drug effects