Comparison Between Tidally Driven Groundwater Flow and Flushing of Animal Burrows in Tropical Mangrove Swamps

Tidal groundwater in a mangrove swamp can return to the mangrove creek by one of two mechanisms: (a) it can either flow through the swamp soil due to the water table difference between the creek and the groundwater in the swamp; or (b) it can flow via tidal flushing of animal burrows. This paper compares the magnitude of these two mechanisms for different regions of a mangrove swamp. Direct groundwater flow rates resulting from water stored in the sediment as a consequence of infiltration, especially during and after tidal inundation, were calculated for every square meter in the surface of a mangrove forest from piezometer data. Flow rates of water due to burrow flushing were determined based on published surveys, by estimating the burrow volume and the percentage of the burrow water that is flushed at each tidal inundation. Although direct groundwater flux was found to decrease further away from the creek compared to close to the creek, it was also found to have a similar range as burrow flushing flow. Specifically, direct groundwater flow ranged from 0.004 to 0.04 m³/m²/day, whilst burrow flushing flux ranged from 0.01 to 0.04 m³/m²/day.Considering the errors involved in the experiments and calculations, these ranges can be considered as being the same and neither of the two processes can be considered as negligible compared to the other. As a consequence, surveys of groundwater processes in mangrove areas, and more generally in swamp and tidal areas where animal burrows are present, will need to consider both mechanisms. Investigations of the influence over flushing mechanisms of different residence times of the water in burrows and in the sediment body would also be recommended in order to establish salt and nutrient budget in mangrove swamps.     

Occurrence of Vibrionaceae in crude and mechanically purified urban sewage; evaluation of their sensitivity to selected drugs

The aim of this study was to ascertain whether bacteria from Vibrionaceae family are present in crude and mechanically cleaned urban sewage and what is their profile of resistance to selected chemotherapeutics. The presence in sewage of bacteria of Vibrionaceae family was proved. They constitute only small percentage of total number of bacteria. No influence of mechanical purification process of sewage on the reduction of total number of bacteria and bacteria from Vibrionaceae family was seen. The number of bacteria from Vibrionaceae family amounted to 6-9 cells per 100 ml of crude sewage and 2-16 cells per 100 ml of mechanically cleaned sewage. From samples tested the following species were isolated: non 01 V. cholerae, V. fluvialis, V. parahaemolyticus, A. hydrophila, A. caviae, A. sobria. All strains were sensitive to neomycin and nalidixic acid and with few exceptions of A. caviae strains to streptomycin, gentamicin, doxycycline and chloramphenicol .     

Fine-scale environmental heterogeneities of tidal creeks affect distribution of crab burrows in a Chinese salt marsh

Tidal creeks are an important structure of salt marshes in estuarine ecosystems, providing valuable ecosystem services to wildlife in the estuary. To determine the effects of environmental heterogeneities within tidal creeks on the features of crab burrows, we divided a typical creek section into four parts (i.e., microhabitats): bottom, slope, edge and flat, investigated the distribution of crab burrows and sediment properties on creek sections in the Yangtze River estuary, and compared the burrow distribution in tidal creeks with that in non-creek areas. Our results showed that from the creek bottom to flat soil water content declined (F_3, 60 = 93.8, p < 0.001), and the variations of other sediment physical and chemical properties associated with the change of soil water content were significant among the microhabitats on the creek sections (p < 0.001 for pH, conductivity, and grain size). No crab burrows were found at the creek bottom. The burrows on the slope were smaller in size (p < 0.001 for burrow opening diameter) while the density was higher than that at the edge and on the flat (F_2, 45 = 31.2, p < 0.001). Moreover, although the correlations between burrow distribution and sediment properties varied among the microhabitats on the creek sections, crabs generally selected relatively solid sediments to build their burrows. On the slope, there was a significantly negative relationship between burrow density and soil water content (r² = 0.53, p < 0.001). At the edge, the correlation between total burrow opening area and soil water content was significantly negative (r² = 0.44, p < 0.002). The density of small crab burrows (<10 mm) was greater, but that of large burrows (>10 mm) was lower in tidal creeks than in non-creek habitats. Therefore, sediment properties showed a gradual transition from hydrophytic to terrestrial environments on the creek section, which caused significant differences of burrow distribution among the microhabitats. The creeks of tidal salt marshes could affect ecological processes and functioning through affecting crab burrows.     

Environmental factors affecting the spawning burrow selection by the gobiid Valenciennea longipinnis

Spawning burrow selection by the longfinned goby Valenciennea longipinnis was studied in the near-shore moat on coral reefs, Okinawa, Japan. The gobies make several burrows within their home range, and spawn in one of them. To examine the factors important for spawning burrow selection three characteristics were investigated: current strength, burrow length and effect of underground water on the burrow. Among the burrows, pairs tended to spawn in a larger burrow irrespective of their body sizes. Most of the other non-spawning burrows were too small for a pair to stay together, because hard substrata may prevent the fish from excavating and shaping the burrow as they like. Moreover, pairs preferred to spawn in burrows where the underground water was oozing out, probably because the male's parental burden will decrease due to the higher dissolved oxygen concentration in such burrows. Although current strength may affect a water-exchange in a V. longipinnis burrow in relation to water-exchange function of a mound, it did not affect the spawning burrow selection because of the smaller velocity difference among the burrows relative to the daily fluctuation of tidal current.     

PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the Gulf of Mexico

This study describes the optimization of PCR parameters and testing of a wide number of microbial species to establish a highly specific and sensitive PCR-based method of detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 strain in pure cultures and seeded waters from the Gulf of Mexico (gulf water). The selected open reading frame 8 (ORF8) DNA-specific oligonucleotide primers tested were found to specifically amplify all 35 pathogenic V. parahaemolyticus O3:K6 pandemic isolates, whereas these primers were not found to detectably amplify two strains of V. parahaemolyticus O3:K6 that were isolated prior to the 1996 outbreaks, 122 non-O3:K6 strains of V. parahaemolyticus, 198 non-V. parahaemolyticus spp., or 16 non-Vibrio bacterial spp. The minimum level of detection by the PCR method was 1 pg of purified genomic DNA or 10(2) ORF8-positive V. parahaemolyticus O3:K6 cells in 100 ml of water. The effectiveness of this method for the detection of ORF8-positive isolates in environmental samples was tested in gulf water seeded with 10-fold serial dilutions of this pathogen. A detection level of 10(3) cells per 100 ml of gulf water was achieved. Also, the applicability of this methodology was tested by the detection of this pathogen in gulf water incubated at various temperatures for 28 days. This PCR approach can potentially be used to monitor with high specificity and well within the required range of sensitivity the occurrence and distribution of this newly emerged pathogenic V. parahaemolyticus O3:K6 strain in coastal, marine, and ship ballast waters. Early detection of V. parahaemolyticus O3:K6 will help increase seafood safety and decrease the risk of infectious outbreaks caused by this pathogen.     

用基于TaqMan探针的Real-time PCR技术定量检测副溶血弧菌

副溶血弧菌是一种引起食源性疾病的重要病原菌,传统的鉴定方法费时费力且容易出现假阴性,建立一种定量检测副溶血弧菌基因的方法尤为重要。根据GenBank公布的副溶血弧菌的gyrB基因序列设计一对引物和TaqMan探针,建立了基于TaqMan探针的RealtimePCR方法。通过对9种细菌(12株菌株)的DNA进行扩增,结果所有4株副溶血弧菌均可产生扩增曲线,其他8株非副溶血弧菌均不产生扩增曲线,证明了引物和探针具有很高的特异性。细菌纯培养物品和人工布菌的检测敏感度分别为1CFUPCR反应体系和10CFUPCR反应体系,相关系数均为0.99(r2=0.99),整个试验可在1h内完成。建立的方法可用于海产品中副溶血弧菌的快速定量检测。     

Comparison of a fluorogenic assay with a conventional method for rapid detection of Vibrio parahaemolyticus in seafoods.

A conventional method and a fluorogenic assay for the detection of Vibrio parahaemolyticus were compared. Among 29 seafood samples examined for the presence of V. parahaemolyticus, 17 samples harbored V. parahaemolyticus, and trypsinlike activity was noticed in 19 seafoods. The added fluorogenic substrate was cleaved in single samples of shrimp, turbo, and cuttlefish from which V. parahaemolyticus could not be isolated by the conventional method. Vibrio alginolyticus, in addition to V. parahaemolyticus, was found to exhibit intracellular trypsinlike activity. Trypsinlike activity in seafoods was observed after the most probable number for the initial density of V. parahaemolyticus-like organisms was found to have reached > 10(2) per g. A V. parahaemolyticus inoculum at 10(4) CFU/ml in arabinose-glucuronate medium was required to attain growth to 10(6) CFU/ml, which is the level necessary for the release of detectable amounts of fluorescent compound from the added substrate.     

Comparative Burrow Architectures of Resident Fiddler Crabs (Ocypodidae) in Indian Sundarban Mangroves to Assess Their Suitability as Bioturbating Agents

Excavation of burrows by fiddler crabs (genus Uca) is an important component in mangrove ecosystem functioning. This bioturbation activity can be measured by analysing the burrow architecture of these crabs. The aim of the present study is to describe and evaluate inter specific differences in the burrow morphologies of four species of fiddler crabs (Uca rosea, Uca triangularis, Uca dussumieri and Uca vocans) using polyester resin casts of the burrows. For each of the species, sex and carapace width (CW; mm) were determined for all the individuals. Three burrow morphological characters viz. burrow diameter (BD; mm), total burrow depth (TBD; mm) and burrow volume (BV; cm³) were considered during the study. Density of each species throughout the year was also assessed. For all the species BD and BV were higher in case of males compared to the females and they showed significant positive correlation with the CW of the burrow inhabitants. The amount of sediment excavated by each crab was evaluated in terms of BV. Among all the studied species, U. rosea was established as the most potent bioturbative candidate in the studied mangrove due to their greater density and moderate ability to excavate burrow.     

Seasonal distribution of facultatively enteropathogenic vibrios (Vibrio cholerae, Vibrio mimicus, Vibrio parahaemolyticus) in the freshwater of the Elbe River at Hamburg

Between June 1981 and December 1982 the incidence of Vibrio cholerae, V. mimicus and V. parahaemolyticus was determined at two sampling sites on the Elbe River at Hamburg. A total of 183 strains was isolated from 147 water samples. Of these, 107 belonged to non-01 V. cholerae (ten strains producing a cholera-like enterotoxin); 33 were identified as V. mimicus, including two enterotoxin producers; 42 strains were Kanagawa-negative cultures of V. parahaemolyticus; and one was V. fluvialis. The highest incidence was observed from June to September with about 10(2) organisms/l. Halophilic vibrios, less than five organisms/l, were detectable during the period June/July to October. The vibrio incidence was not influenced by the numbers of aerobic heterotrophic bacteria, coliforms or faecal bacteria. In general water temperature correlated with the seasonal variation. Thus, a temperature rise over 10 degrees to 20 degrees C was followed by a distinct increase in vibrio numbers. Of 14 chemical parameters only chloride concentration might have had an influence on the seasonal variation. It is concluded that the three Vibrio species are indigenous organisms of the Elbe River.     

Seasonal distribution of facultatively enteropathogenic vibrios (Vibrio cholerae, Vibrio mimicus, Vibrio parahaemolyticus) in the freshwater of the Elbe River at Hamburg

Between June 1981 and December 1982 the incidence of Vibrio cholerae, V. mimicus and V. parahaemolyticus was determined at two sampling sites on the Elbe River at Hamburg. A total of 183 strains was isolated from 147 water samples. Of these, 107 belonged to non-01 V. cholerae (ten strains producing a cholera-like enterotoxin); 33 were identified as V. mimicus , including two enterotoxin producers; 42 strains were Kanagawa-negative cultures of V. parahaemolyticus ; and one was V. fluvialis. The highest incidence was observed from June to September with about 10² organisms/1. Halophilic vibrios, less than five organisms/1, were detectable during the period June/July to October. The vibrio incidence was not influenced by the numbers of aerobic heterotrophic bacteria, coliforms or faecal bacteria. In general water temperature correlated with the seasonal variation. Thus, a temperature rise over 10° to 20°C was followed by a distinct increase in vibrio numbers. Of 14 chemical parameters only chloride concentration might have had an influence on the seasonal variation. It is concluded that the three Vibrio species are indigenous organisms of the Elbe River.     

Behavior of the groundwater in a riverine-type mangrove forest

The behavior of groundwater and physical properties of bottom sediment in a riverine-type mangrove forest which is composed of a tidal creek and fringing mangrove swamps were investigated through field observations at Iriomote Island, Japan. After the tidal water ebbed from the swamp surface to the creek, groundwater levels at swamp sites near the creek fell by up to 15 cm by the next flood tide, although the fall was negligible at sites far from the creek and at the open coast outside the mangrove forest. The amount of groundwater discharged to the creek from the swamp depended strongly both on the tidal range and the presence of the steep bank which separates the tidal creek from the fringing mangrove swamp. Based on the fall of groundwater level, the bulk hydraulic conductivity of the swamp was estimated to be 1.5×10⁻² cm/s. This value is two to three orders of magnitude larger than that measured in a laboratory using small scale sediment core samples collected in the swamp. These results suggest that the presence of crowded, intricate and large animal burrows as well as sediment layers rich in mangrove humus increases permeability in the mangrove swamp. Further, it is suggested that the mangrove topography with the steep bank of the tidal creek plays an important role which enhances material exchanges through groundwater between the mangrove swamp and the adjacent offshore waters.     

Development of a multiplex real-time PCR assay with an internal amplification control for the detection of total and pathogenic Vibrio parahaemolyticus bacteria in oysters

Vibrio parahaemolyticus is an estuarine bacterium that is the leading cause of shellfish-associated cases of bacterial gastroenteritis in the United States. Our laboratory developed a real-time multiplex PCR assay for the simultaneous detection of the thermolabile hemolysin (tlh), thermostable direct hemolysin (tdh), and thermostable-related hemolysin (trh) genes of V. parahaemolyticus. The tlh gene is a species-specific marker, while the tdh and trh genes are pathogenicity markers. An internal amplification control (IAC) was incorporated to ensure PCR integrity and eliminate false-negative reporting. The assay was tested for specificity against >150 strains representing eight bacterial species. Only V. parahaemolyticus strains possessing the appropriate target genes generated a fluorescent signal, except for a late tdh signal generated by three strains of V. hollisae. The multiplex assay detected <10 CFU/reaction of pathogenic V. parahaemolyticus in the presence of >10(4) CFU/reaction of total V. parahaemolyticus bacteria. The real-time PCR assay was utilized with a most-probable-number format, and its results were compared to standard V. parahaemolyticus isolation methodology during an environmental survey of Alaskan oysters. The IAC was occasionally inhibited by the oyster matrix, and this usually corresponded to negative results for V. parahaemolyticus targets. V. parahaemolyticus tlh, tdh, and trh were detected in 44, 44, and 52% of the oyster samples, respectively. V. parahaemolyticus was isolated from 33% of the samples, and tdh(+) and trh(+) strains were isolated from 19 and 26%, respectively. These results demonstrate the utility of the real-time PCR assay in environmental surveys and its possible application to outbreak investigations for the detection of total and pathogenic V. parahaemolyticus.     

Analysis of gyrB and toxR gene sequences of Vibrio hollisae and development of gyrB- and toxR-targeted PCR methods for isolation of V. hollisae from the environment and its identification

Isolation of Vibrio hollisae strains, particularly from the environment, is rare. This may be due, in part, to the difficulty encountered when using conventional biochemical tests to identify the microorganism. In this study, we evaluated whether two particular genes may be useful for the identification of V. hollisae. The two genes are presumed to be conserved among the bacterial species (gyrB) or among the species of the genus Vibrio (toxR). A portion of the gyrB sequence of V. hollisae was cloned by PCR using a set of degenerate primers. The sequence showed 80% identity with the corresponding Vibrio parahaemolyticus gyrB sequence. The toxR gene of V. hollisae was cloned utilizing a htpG gene probe derived from the V. parahaemolyticus htpG gene, which is known to be linked to the toxR gene in V. hollisae. The coding sequence of the cloned V. hollisae toxR gene had 59% identity with the V. parahaemolyticus toxR coding sequence. The results of DNA colony hybridization tests using the DNA probes derived from the two genes of V. hollisae indicated that these gene sequences could be utilized for differentiation of V. hollisae from other Vibrio species and from microorganisms found in marine fish. PCR methods targeting the two gene sequences were established. Both PCR methods were shown to specifically detect the respective target sequences of V. hollisae but not other organisms. A strain of V. hollisae added at a concentration of 1 to 10(2) CFU/ml to alkaline peptone water containing a seafood sample could be detected by a 4-h enrichment incubation in alkaline peptone water at 37 degrees C followed by quick DNA extraction with an extraction kit and 35-cycle PCR specific for the V. hollisae toxR gene. We conclude that screening of seafood samples by this 35-cycle, V. hollisae toxR-specific PCR, followed by isolation on a differential medium and identification by the above htpG- and toxR-targeted PCR methods, can be useful for isolation from the environment and identification of V. hollisae.     

Effect of Mg(2+) ion in protein secretion by magnesium-resistant strains of Pseudomonas aeruginosa and Vibrio parahaemolyticus isolated from the coastal water of Haldia port

A rapidly growing industrial complex including oil refineries and chemical industries has developed around the coastal area of Haldia port in the district of Midnapore, West Bengal, India. The coastal water is highly polluted with industrial wastes along with petroleum hydrocarbons. The bacteria isolated from the different sites of the coastal waters were Escherichia coli, Alcaligenes, Acinetobacter, Klebsiella spp., Micrococcus spp., Vibrio spp., Pseudomonas aeruginosa and Vibrio parahaemolyticus. The salinity of the water during the time of collection of samples around the port area was 8. 2 ppt. Among the isolated organisms, only two isolates, P. aeruginosa and V. parahaemolyticus, showed growth at 300 mM Mg(2+) ion concentration. However, a 3 mM Mg(2+) concentration was detected in the coastal water whereas other metal ion concentrations were less than 3x10(-5) mM. Resistance to Mg(2+) (300 mM) was determined by a 5.5-kb plasmid. A large amount of a 40-kDa outer membrane protein, which was highly soluble in 1 M MgCl(2), was isolated from both V. parahaemolyticus and P. aeruginosa. The secretion of proteins in the culture supernatant of V. parahaemolyticus was highly increased when the cells were grown in the presence of 300 mM Mg(2+), whereas very low secretion was observed in the same concentration of Mg(2+) in the case of P. aeruginosa. Mg(2+) may act as a specific release factor in protein secretion by V. parahaemolyticus strains.     

Identification of Vibrio harveyi using PCR amplification of the toxR gene

AIMS: The aim of this study was to develop an effective method for the identification of Vibrio harveyi based on using the toxR gene as a taxonomic marker. METHODS AND RESULTS: Primers for the toxR gene were designed for specificity to V. harveyi, and incorporated in a polymerase chain reaction (PCR). The results of the PCR, which took <5 h from DNA extraction to amplification, revealed positive amplification of the toxR gene fragment in 20 V. harveyi isolates including type strains, whereas DNA from 23 other Vibrionaceae type strains and 13 Vibrio parahaemolyticus strains were negative. The detection limit of the PCR was 4.0 x 10(3) cells ml(-1). In addition, the technique enabled the recognition of V. harveyi from diseased fish. CONCLUSIONS: The PCR was specific and sensitive, enabling the identification of V. harveyi within 5 h. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR allowed the rapid and sensitive detection of V. harveyi.     

Serological method for identification of Vibrio parahaemolyticus from marine samples

Use of agglutination with antiserum against lateral flagella (H-agglutination) for the identification of Vibrio parahaemolyticus was studied. Sucrose-negative bacteria were isolated from seawater, and their characterization was carried out by traditional biological tests and slide agglutination with antiserum specific to lateral flagella of V. parahaemolyticus. Of 135 strains isolated, 78 were identified as V. parahaemolyticus by biological tests and were agglutinated with the above serum. Fifty-five strains did not agglutinate with the serum, and their biological characteristics were different from those of V. parahaemolyticus. Two strains also differed from V. parahaemolyticus in some biological characteristics but agglutinated with the antiserum. All clinically isolated V. parahaemolyticus strains also agglutinated with the above serum. These results suggest that our serological method is useful for the identification of V. parahaemolyticus, especially for samples in which there are many organisms related to V. parahaemolyticus, because many biological tests can be omitted.     

Serological method for identification of Vibrio parahaemolyticus from marine samples.

Use of agglutination with antiserum against lateral flagella (H-agglutination) for the identification of Vibrio parahaemolyticus was studied. Sucrose-negative bacteria were isolated from seawater, and their characterization was carried out by traditional biological tests and slide agglutination with antiserum specific to lateral flagella of V. parahaemolyticus. Of 135 strains isolated, 78 were identified as V. parahaemolyticus by biological tests and were agglutinated with the above serum. Fifty-five strains did not agglutinate with the serum, and their biological characteristics were different from those of V. parahaemolyticus. Two strains also differed from V. parahaemolyticus in some biological characteristics but agglutinated with the antiserum. All clinically isolated V. parahaemolyticus strains also agglutinated with the above serum. These results suggest that our serological method is useful for the identification of V. parahaemolyticus, especially for samples in which there are many organisms related to V. parahaemolyticus, because many biological tests can be omitted.     

Temporal and spatial variability in culturable pathogenic Vibrio spp. in Lake Pontchartrain, Louisiana, following hurricanes Katrina and Rita

We investigated the abundance, distribution, and virulence gene content of Vibrio cholerae, V. parahaemolyticus, and V. vulnificus in the waters of southern Lake Pontchartrain in Louisiana on four occasions from October 2005 to September 2006, using selective cultivation and molecular assays. The three targeted pathogenic vibrios were generally below the detection level in January 2006, when the water was cold (13°C), and most abundant in September 2006, when the lake water was warmest (30°C). The maximum values for these species were higher than reported previously for the lake by severalfold to orders of magnitude. The only variable consistently correlated with total vibrio abundance within a single sampling was distance from shore (P = 0.000). Multiple linear regression of the entire data set revealed that distance from shore, temperature, and turbidity together explained 82.1% of the variability in total vibrio CFU. The log-transformed mean abundance of V. vulnificus CFU in the lake was significantly correlated with temperature (P = 0.014), but not salinity (P = 0.625). Virulence-associated genes of V. cholerae (ctx) and V. parahaemolyticus (trh and tdh) were not detected in any isolates of these species (n = 128 and n = 20, respectively). In contrast, 16S rRNA typing of V. vulnificus (n = 298) revealed the presence of both environmental (type A) and clinical (type B) strains. The percentage of the B-type V. vulnificus was significantly higher in the lake in October 2005 (35.8% of the total) than at other sampling times (P ≤ 0.004), consistent with the view that these strains represent distinct ecotypes.     

Homing in fiddler crabs (Uca lactea annulipes and Uca vomeris : Ocypodidae)

Fiddler crabs emerge from burrows on intertidal sand- and mudflats to feed during low tide. In the species studied here (Uca lactea annulipes, Uca vomeris) a crab normally wanders no more than about 1 m away from its burrow and, when frightened, dashes back along a straight line to take cover. Feeding crabs tend to move sideways, without changing orientation, along paths radiating from the burrow. When they move along circumferential paths they adjust their orientation so that one side continues to point towards the burrow. The crabs do not need to see the burrow in order to stay aligned with the home vector, and they are not misled by a dummy hole close to their own burrow unless they have come to within about 10 cm of it. The home runs of crabs end within a few centimeters of a burrow that is covered with a sheet of sandpaper and then give way to search runs, centred upon a position slightly short of the burrow location. Feeding crabs can be displaced on sandpapers and their subsequent home runs end at a position where the burrow would be, had there been no displacement. Landmarks close to the burrow do not influence the home runs of displaced crabs. Crabs that are rotated on a sheet of sandpaper, counter-turn to keep their original orientation constant. Fiddler crabs thus employ path integration with external compass information and close range visual guidance for homing. Accepted: 11 May 1998     

Application of real-time PCR for quantitative detection of Vibrio parahaemolyticus from seafood in eastern China

Vibrio parahaemolyticus is recognized as a leading human food-borne pathogen. A TaqMan PCR assay based on the gyrase B gene (gyrB) sequence of V. parahaemolyticus was developed for quantitative detection of V. parahaemolyticus in seafood. The study involving 27 V. parahaemolyticus and 10 strains of other species indicated that the real-time PCR test was highly specific. The sensitivity of the assay was approximately a single CFU per PCR in pure culture and six to eight CFU per PCR in spiked raw oyster, respectively. Real-time PCR values of artificially inoculated oyster homogenates correlated well with plate counts determined using culture methods. A total of 300 seafood samples were analyzed and 78 (26%) of these samples were positive for V. parahaemolyticus using a conventional culture method and 97 (32.3%) using the real-time PCR assay. All culture-positive samples were PCR-positive. However, 19 samples positive by PCR were culture-negative. The results show that retail seafood is commonly contaminated with V. parahaemolyticus in harvest season in eastern China. These data also indicate that real-time PCR can provide sensitive species-specific detection and quantification of V. parahaemolyticus in seafood without prior isolation and characterization of the bacteria by traditional microbiological methods.