scholarly journals First Report of a Root and Crown Disease Caused by Rhizoctonia solani on Centaurea stoebe in Russia

Plant Disease ◽  
2009 ◽  
Vol 93 (12) ◽  
pp. 1350-1350
Author(s):  
A. J. Caesar ◽  
R. T. Lartey ◽  
T. Caesar-TonThat
Keyword(s):  
Rhizoctonia Solani ◽  
Plant Pathogens ◽  
The United States ◽  
Economic Losses ◽  
Most Probable Number ◽  
Infested Soil ◽  
Spotted Knapweed ◽  
Centaurea Stoebe ◽  
Probable Number ◽  
First Report

Spotted knapweed (SKW), Centaurea stoebe L., is a nonindigenous species that is invasive over large areas in the United States, especially in the west. It has been estimated that infestations of SKW cause $42 million in direct and indirect economic losses annually (2), and the weed could potentially invade 13.6 million ha of rangeland in Montana alone. Extensive efforts toward the control of SKW have included the release of 12 insects for biological control, four of which attack the crowns and roots of this short-lived perennial. To focus efforts to select potential soilborne pathogens, which could be applied in combination with insects, we conducted a survey for plant pathogens in the native range of SKW associated with damage caused by any root-attacking insects. Stunted and chlorotic SKW plants, which were colonized by larvae of Cyphocleonus spp., were found in June 1994 near the Novomar'evskaya Botanical Sanctuary (45°08′49.87″N, 41°51′02.05″E) in the Caucasus Region of Russia. A nonsporulating multinucleate fungus was isolated from the lower stem, crown, and upper root tissue of one such plant. Colonies growing on potato dextrose agar and Ko and Hora media were examined microscopically and identified as Rhizoctonia solani by the occurrence of robust, thick-walled, golden hyphae with right-angled branching and constrictions at the branch points. The anastomosis grouping of the one isolate was determined to be AG 2-2 IIIB after pairing it on water agar with 11 AG tester isolates representing all subgroups of AG 1 to AG 5. The hyphal diameter at the obvious point of anastomosis was reduced and cell death of adjacent cells was observed. In 2007, pathogenicity was determined by planting 12-week-old seedlings of SKW, one per pot, into 20 15-cm-diameter pots of a steamed greenhouse soil mix composed of sphagnum peat, sand, and Bozeman silt loam (1:1:1, vol/vol), pH 6.6, infested with R. solani-colonized barley grain that had been dried and milled. An inoculum level of 8 CFU/g of air-dried soil was determined by most probable number calculations from fourfold dilutions of infested soil. Controls were planted into noninfested soil. In both greenhouse tests, the isolate caused either mortality or a 93% mean fresh weight reduction of surviving plants, relative to the controls, after 8 months. R. solani was reisolated from necrotic root and crown tissue of dead and stunted plants but not from the controls. To our knowledge, this is the first report of R. solani occurring on SKW in Europe. The characterization and pathogenicity of Fusarium spp. isolated from insect-colonized roots of SKW in Europe was reported previously (1). References: (1) A. J. Caesar et al. BioControl 47:217. (2) S. A. Hirsch and J. A. Leitch, North Dakota Agricultural Economics Report No. 355. NDSU, Fargo. 1996.

scholarly journals First Report of Rhizoctonia spp. Causing a Root Rot of the Invasive Rangeland Weed Lepidium draba in North America

Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1278-1278 ◽  
Author(s):  
A. J. Caesar ◽  
R. T. Lartey ◽  
T. Caesar-TonThat ◽  
J. Gaskin
Keyword(s):  
North America ◽  
Native Species ◽  
The United States ◽  
Root Tissue ◽  
Most Probable Number ◽  
Infested Soil ◽  
Native Range ◽  
Probable Number ◽  
First Report ◽  
Lepidium Draba

The exotic, invasive perennial rangeland weed Lepidium draba spreads rapidly and reduces native species diversity. The extensive root system of L. draba constitutes 76% of its biomass (4). Thus, searches have been done for biocontrol agents that target root tissue or that may interact with a weevil, Ceutorhynchus assimilis, that causes galls in the crown area of L. draba. An association of Rhizoctonia spp. with root tissue of plants galled by the weevil has been documented in Europe (3). The possible presence of soilborne pathogens similar to those found in the native range has been the subject of L. draba surveys in the United States. One such survey in 2008 detected a few plants with reddened and chlorotic foliage in a stand near Shepherd, MT. Such symptoms typically indicate the occurrence of soilborne diseases on L. draba in the native range of the weed (2). The site had shown a gradual increase in the range of detectable pathogens beginning with foliar pathogens in 1997. In 2010, at the Shepherd site, L. draba plants with similar (but more severe) symptoms to those seen in 2008 were noted in a different area of the stand. Excavation of the roots in both years revealed brown, sunken crown and root cankers. Pieces of root tissue were excised from the lesions and plated on acidified PDA and Ko and Hora medium. A non-sporulating fungus was isolated from three plants. Colonies of the isolates on PDA were typical of known Rhizoctonia spp. The 2010 isolates were determined to be multinucleate using DAPI and were paired with 14 tester (including subgroups) isolates of AG-1 to AG-4 on water agar. Anastomosis was observed between the multinucleate isolates and the AG-2-1 tester isolate. Sequence analysis of ITS of the rDNA of a multinucleate isolate (GenBank KJ545577) indicated 99% similarity with an accession of R. solani AG 2-1 (AB547381). The 2008 isolates were binucleate. A binucleate isolate, KJ545578, had 100% similarity with an isolate of Rhizoctonia spp. AG-A (AY927356). Pathogenicity tests consisted of planting 6-week-old seedlings of L. draba, one per pot, in ten 85-cm-diameter pots of pasteurized soil mix infested with Rhizoctonia-colonized barley grain that had been dried and milled. An inoculum level of ~8 CFU/g (1) of air-dried soil was established by most probable number calculations from fourfold dilutions of infested soil. Controls were the same number of plants in pasteurized potting mix. Results were recorded after 3 months in a greenhouse at 20–25°C. The test was repeated. Typically, R. solani caused mortality of six to eight plants, from which it was re-isolated, whereas binuclate isolates caused stunting and lower dry weight of L. draba. Control plants remained asymptomatic. This is the first report of R. solani and binucleate Rhizoctonia spp. on L. draba in North America. References: (1) A. J. Caesar et al. Plant Dis. 93:1350, 2009. (2) A. J. Caesar et al. Biol. Control 52:140, 2010. (3) A. J. Caesar et al. Plant Dis. 96:145, 2011. (4) R. F. Miller et al. Agronomy J. 86:487, 1994.

scholarly journals First Report of Damping-Off of Swiss Chard Caused by Rhizoctonia solani AG-4 HG I and Binucleate Rhizoctonia AG-A in China

Plant Disease ◽  
2007 ◽  
Vol 91 (11) ◽  
pp. 1516-1516 ◽  
Author(s):  
G. H. Yang ◽  
R. L. Conner ◽  
Y. Y. Chen
Keyword(s):  
Rhizoctonia Solani ◽  
Root Zone ◽  
The United States ◽  
Infested Soil ◽  
Damping Off ◽  
First Report ◽  
Binucleate Rhizoctonia ◽  
5.8S Rdna ◽  
Stem Blight ◽  
Swiss Chard

During July, 2003, damping-off of Swiss chard (Beta vulgaris subsp. cicla L.) was observed in a seedling (approximately 1 month after germination) field (approximately 2 ha) in Yuanmou County in the Cuxiong District of Yunnan, China. More than 80% of the seedlings showed symptoms of the disease. Symptoms on newly emerged plants consisted of wilting, a brown necrosis of the lower taproot, and eventual death of seedlings. Among the 15 isolates of Rhizoctonia spp. isolated from Swiss chard with damping-off symptoms, 12 isolates of Rhizoctonia solani with dark brown sclerotia on potato dextrose agar (PDA) anastomosed with tester isolates of each subgroup AG-4 HG I, AG-4 HG II, and AG-4 HG III, giving a C2 hyphal fusion (1) reaction at a high frequency. The other three binucleate Rhizoctonia spp. (BNR) isolates whose mycelia were white with floccose aerial hyphae on PDA anastomosed freely with two BNR AG-A tester isolates producing a C2 hyphal reaction. The 5.8S rDNA-ITS of a single isolate of R. solani and a single isolate of BNR was sequenced. The sequence of the AG-4 isolate (GenBank Accession No. EF679777) exhibited 99 to 100% homology with isolates of R. solani AG-4, subgroup 4HG I (GenBank Accession No. AY154307). The sequence from the AG-A isolate (GenBank Accession No. EF679778) exhibited 98% homology with BNR AG-A (GenBank Accession Nos. AB000040 and AF354092). Swiss chard (cv. Baijin) seedlings (approximately 5 cm high) were planted in potting soil at a density of one seedling per vinyl pot (8 cm diameter, 9 cm high). Two isolates each of R. solani and BNR were used in pathogenicity testing. Each seedling was inoculated in the root zone with approximately 7 g of artificially infested soil. Control plants were inoculated with autoclaved soil. The experiments were conducted three times, each time with three replicates, in a greenhouse with a photoperiod of 16 h of light and 8 of h dark at 30 and 16°C, respectively. After 7 days, disease severity was measured based on a scale in which 0 = no symptom; 1 = small lesions on seedlings, no blight; 2 = leaves blight, no stem blight; 3 = stem blight; and 4 = plant dead. The two AG-4 and two of AG-A isolates were pathogenic on the Swiss chard seedlings and caused damping-off symptoms with a disease index 1.7 to 4.0, and there were no significant differences (P = 0.05) among them. We reisolated and confirmed the presence of R. solani and BNR AG-A from diseased plants. AG-3 isolates were reported to cause the damping-off of Swiss chard in the United States (2). To our knowledge, this is the first report of damping-off of Swiss chard caused by Rhizoctonia solani AG-4 HG I and BNR AG-A. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reaction. Kluwer Academic Publishers, Dordecht, the Netherlands, 1996. (2) S. T. Koike and K. V. Subbarao. Plant Dis. 83:695, 1999.

scholarly journals First Report of Impatiens Downy Mildew Outbreaks Caused by Plasmopara obducens Throughout the Hawai'ian Islands

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 696-696 ◽  
Author(s):  
J. A. Crouch ◽  
M. P. Ko ◽  
J. M. McKemy
Keyword(s):  
United States ◽  
Downy Mildew ◽  
The United States ◽  
Molecular Data ◽  
Economic Losses ◽  
Voucher Specimen ◽  
First Report ◽  
Plastic Bags ◽  
Leaf Surfaces ◽  
Impatiens Walleriana

Downy mildew of impatiens (Impatiens walleriana Hook.f.) was first reported from the continental United States in 2004. In 2011 to 2012, severe and widespread outbreaks were documented across the United States mainland, resulting in considerable economic losses. On May 5, 2013, downy mildew disease symptoms were observed from I. walleriana ‘Super Elfin’ at a retail nursery in Mililani, on the Hawai'ian island of Oahu. Throughout May and June 2013, additional sightings of the disease were documented from the islands of Oahu, Kauai, Maui, and Hawai'i from nurseries, home gardens, and botanical park and landscape plantings. Symptoms of infected plants initially showed downward leaf curl, followed by a stippled chlorotic appearance on the adaxial leaf surfaces. Abaxial leaf surfaces were covered with a layer of white mycelia. Affected plants exhibited defoliation, flower drop, and stem rot as the disease progressed. Based on morphological and molecular data, the organism was identified as Plasmopara obducens (J. Schröt.) J. Schröt. Microscopic observation disclosed coenocytic mycelium and hyaline, thin-walled, tree-like (monopodial branches), straight, 94.0 to 300.0 × 3.2 to 10.8 μm sporangiophores. Ovoid, hyaline sporangia measuring 11.0 to 14.6 × 12.2 to 16.2 (average 13.2 × 14.7) μm were borne on sterigma tips of rigid branchlets (8.0 to 15.0 μm) at right angle to the main axis of the sporangiophores (1,3). Molecular identification of the pathogen was conducted by removing hyphae from the surface of three heavily infected leaves using sterile tweezers, then extracting DNA using the QIAGEN Plant DNA kit (QIAGEN, Gaithersburg, MD). The nuclear rDNA internal transcribed spacer was sequenced from each of the three samples bidirectionally from Illustra EXOStar (GE Healthcare, Piscataway, NJ) purified amplicon generated from primers ITS1-O and LR-0R (4). Resultant sequences (GenBank KF366378 to 80) shared 99 to 100% nucleotide identity with P. obducens accession DQ665666 (4). A voucher specimen (BPI892676) was deposited in the U.S. National Fungus Collections, Beltsville, MD. Pathogenicity tests were performed by spraying 6-week-old impatiens plants (I. walleriana var. Super Elfin) grown singly in 4-inch pots with a suspension of 1 × 104 P. obducens sporangia/ml until runoff using a handheld atomizer. Control plants were sprayed with distilled water. The plants were kept in high humidity by covering with black plastic bags for 48 h at 20°C, and then maintained in the greenhouse (night/day temperature of 20/24°C). The first symptoms (downward curling and chlorotic stippling of leaves) and sporulation of the pathogen on under-leaf surfaces of the inoculated plants appeared at 10 days and 21 days after inoculation, respectively. Control plants remained healthy. Morphological features and measurements matched those of the original inoculum, thus fulfilling Koch's postulates. To our knowledge, this is the first report of downy mildew on I. walleriana in Hawai'i (2). The disease appears to be widespread throughout the islands and is likely to cause considerable losses in Hawai'ian landscapes and production settings. References: (1) O. Constantinescu. Mycologia 83:473, 1991. (2) D. F. Farr and A. Y. Rossman. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ July 16, 2013. (3) P. A. Saccardo. Syllogue Fungorum 7:242, 1888. (4) M. Thines. Fungal Genet Biol 44:199, 2007.

scholarly journals First Report of Okra yellow mosaic Mexico virus in Okra in the United States

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 924-924 ◽  
Author(s):  
C. Hernandez-Zepeda ◽  
T. Isakeit ◽  
A. Scott ◽  
J. K. Brown
Keyword(s):  
United States ◽  
The United States ◽  
Full Length ◽  
Bipartite Begomoviruses ◽  
Economic Losses ◽  
Growth Stages ◽  
First Report ◽  
Total Dna ◽  
Hidalgo County ◽  
Whitefly Vector

During the okra growing season from August to November of 2009, symptoms reminiscent of geminivirus infection were observed on 75% of ‘Green Emerald’ Abelmoschus esculentus (L.) Moench, plants in a 0.2-km2 field in Hidalgo County, TX. Visible symptoms consisted of irregular yellow patches on leaves, distinctive yellow borders on leaf edges, and chlorosis of subsequently developing leaves. The whitefly vector of begomoviruses, Bemisia tabaci (Genn.), infested okra plants in the early growth stages during late July 2009. Total DNA was isolated from the leaves of three symptomatic okra plant samples (1) and used as the PCR template to amplify a 575-bp fragment of the coat protein gene (CP) using the universal begomovirus primers AV494 and AC1048 (2). PCR products of the expected size were cloned into the pGEM-T Easy (Promega, Madison, WI) and sequenced using the universal M13F and M13 R primers. ClustalV alignment indicated 99 to 100% shared nucleotide (nt) identity, and BLAST analysis revealed that the closest relative was Okra yellow mosaic Mexico virus - Tetekalitla (OkYMMV) (GenBank Accession No. EF591631) at 98%. To amplify the full-length DNA-A and a possible cognate DNA-B component, one plant that was positive by CP-PCR and DNA sequencing was selected for further analysis. Total DNA from this plant was used as template for a second detection method that consisted of rolling circle amplification (RCA) using the TempliPhi 100 Amplification System (GE Healthcare). RCA is a non-sequence-specific approach that permits amplification of circular DNA. The RCA products were linearized to release unit length ~2.6 kb DNA-A and DNA-B components using BamHI, and EcoRI, respectively. These products were cloned into pGEM3zf+ (Promega) and sequenced using M13F and M13 R primers and then by primer walking (>300 base overlap). Full-length DNA-A and DNA-B components were obtained, respectively, at 2,613 bp (GenBank Accession No. HM035059) and 2,594 bp (GenBank Accession No HM035060). Alignment of the DNA-A component using ClustalV (MegAlign, DNASTAR, Madison, WI) with begomoviral sequences available in GenBank indicated that it was 99% identical to OkYMMV DNA-A (GenBank Accession No. DQ022611). The closest relative to the DNA-B component (ClustalV) was Sida golden mosaic virus (SiGMV) (GenBank Accession No. AJ250731) at 73%. The nt identity of the 172-nt ‘common region’ present in the DNA-A and DNA-B components was 99%, and the iterons (predicted Rep binding motif) were identical for the two components, indicating that they are a cognate pair. The genome organization was typical of other New World bipartite begomoviruses. The economic losses due to infection by this virus could not be determined because an early freeze killed the plants. Hidalgo County is adjacent to Tamaulipas, Mexico, where ~50 km2 of okra are grown and the whitefly vector is also present. The identification of OkYMMV based on two independent detection methods, and the presence of begomovirus-like symptoms together with the whitefly vector, provide robust evidence for the association of OkYMMV-TX with diseased okra plants. To our knowledge, this is the first report of OkYMMV-TX infecting okra crops in Texas and in the continental United States. References: (1) J. J. Doyle and J. L. Doyle. Focus 12:13, 1990. (2) S. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.

scholarly journals Fungicide SYP-14288 Inducing Multidrug Resistance in Rhizoctonia solani

Plant Disease ◽  
2020 ◽  
Vol 104 (10) ◽  
pp. 2563-2570 ◽  
Author(s):  
Xingkai Cheng ◽  
Xuejing Man ◽  
Zitong Wang ◽  
Li Liang ◽  
Fan Zhang ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Rhizoctonia Solani ◽  
Parental Strain ◽  
Crop Production ◽  
Plant Pathogens ◽  
Economic Losses ◽  
Cross Resistance ◽  
Baseline Sensitivity ◽  
Rice Plants ◽  
Type Strains

Rhizoctonia solani is a widely distributed soilborne plant pathogen, and can cause significant economic losses to crop production. In chemical controls, SYP-14288 is highly effective against plant pathogens, including R. solani. To examine the sensitivity to SYP-14288, 112 R. solani isolates were collected from infected rice plants. An established baseline sensitivity showed that values of effective concentration for 50% growth inhibition (EC50) ranged from 0.0003 to 0.0138 μg/ml, with an average of 0.0055 ± 0.0030 μg/ml. The frequency distribution of the EC50 was unimodal and the range of variation factor (the ratio of maximal over minimal EC50) was 46.03, indicating that all wild-type strains were sensitive to SYP-14288. To examine the risk of fungicide resistance, 20 SYP-14288-resistant mutants were generated on agar plates amended with SYP-14288. Eighteen mutants remained resistant after 10 transfers, and their fitness was significantly different from the parental strain. All of the mutants grew more slowly but showed high virulence to rice plants, though lower than the parental strain. A cross-resistance assay demonstrated that there was a positive correlation between SYP-14288 and fungicides having or not having the same mode of action with SYP-14288, including fluazinam, fentin chloride, fludioxonil, difenoconazole, cyazofamid, chlorothalonil, and 2,4-dinitrophen. This result showed a multidrug resistance induced by SYP-14288, which could be a concern in increasing the spectrum of resistance in R. solani to commonly used fungicides.

Fecal Coliform Methods for Examination of Sea Water: Interlaboratory Evaluation of Split Sample Analysis

1978 ◽  
Vol 61 (4) ◽  
pp. 772-778
Author(s):  
John J Miescier ◽  
Virgil E Carr ◽  
John F Musselman ◽  
Santo A Furfari
Keyword(s):  
Elevated Temperature ◽  
Sea Water ◽  
The United States ◽  
Most Probable Number ◽  
Test Procedures ◽  
Probable Number ◽  
E Coli ◽  
Split Sample ◽  
Elevated Temperature Test ◽  
Temperature Test

Abstract An interlaboratory study was conducted to compare the effectiveness of the following 3 multiple-tube fermentation methods for determining the most probable number (MPN) of Escherichia coli in a split artificial sea water sample: ( 1 ) the 72-hr standard methods procedure of the American Public Health Association, (2) a 24-hr elevated-temperature test using A-l medium, and (3) a 24-hr elevated temperature test modified to include an initial 3-hr resuscitation period using A-l medium. The capability of the laboratories to perform the 3 test procedures was also compared. Split sample replicates with low, medium, and high levels of E. coli were examined in 18 laboratories in the United States and Canada. Data indicate that the laboratories performed each test with equal capability, and all 3 procedures were equally effective in enumerating the strain of E. coli used in this investigation. By virtue of its homogeneity and stability, the split sample served as an appropriate specimen for this study and could probably be used as a proficiency test specimen for evaluating laboratory analyst performance in the bacteriological examination of sea water.

Survey of Postharvest-Processed Oysters in the United States for Levels of Vibrio vulnificus and Vibrio parahaemolyticus

2009 ◽  
Vol 72 (10) ◽  
pp. 2110-2113 ◽  
Author(s):  
ANGELO DePAOLA ◽  
JESSICA L. JONES ◽  
KATHY E. NOE ◽  
ROBIN H. BYARS ◽  
JOHN C. BOWERS
Keyword(s):  
Vibrio Vulnificus ◽  
Gulf Coast ◽  
Virulence Gene ◽  
Selective Advantage ◽  
The United States ◽  
Most Probable Number ◽  
Probable Number ◽  
Mild Heat ◽  
Thermostable Direct Hemolysin ◽  
The Mean

From June through October 2004, the U.S. Food and Drug Administration collected oysters (61 samples) that had been subjected to postharvest processing (PHP) methods, including mild heat treatment, freezing, and high hydrostatic pressure, from processors and retail markets in various states to determine Vibrio vulnificus and V. parahaemolyticus levels. Presence in a 25-g sample and most probable number (MPN) using standard enrichment and selective isolation procedures were utilized. Suspect colonies were isolated and identified using DNA probe colony hybridization. Neither species of vibrio was detected in 25-g portions of most samples regardless of the PHP. The lowest frequency of isolation of either pathogen (<10%) was observed with the mild heat process. Few (12 to 13%) frozen samples collected at the processor but not at retail contained >30 MPN/g of either pathogen. The mean levels of either organism in PHP oysters observed in the present study were 5 to 6 log less than in unprocessed raw Gulf Coast oysters. Of the 70 V. vulnificus isolates examined, only 5 possessed the putative virulence marker, type B 16S rRNA. Neither the thermostable direct hemolysin (tdh) nor the tdh-related hemolysin (trh) virulence gene was detected in any of the 40 V. parahaemolyticus isolates examined in the present study. These data suggest that if there is any selective advantage to pathogenic strains of V. vulnificus and V. parahaemolyticus, these differences are minimal. These results indicate that all PHP treatments greatly reduce exposure of V. vulnificus and V. parahaemolyticus to raw-oyster consumers. Consequently, these PHP oysters pose a much lower risk of illness to consumers due to these pathogens.

scholarly journals Prevalence of Pandemic Thermostable Direct Hemolysin-Producing Vibrio parahaemolyticus O3:K6 in Seafood and the Coastal Environment in Japan

2003 ◽  
Vol 69 (7) ◽  
pp. 3883-3891 ◽  
Author(s):  
Yukiko Hara-Kudo ◽  
Kanji Sugiyama ◽  
Mitsuaki Nishibuchi ◽  
Ashrafuzzaman Chowdhury ◽  
Jun Yatsuyanagi ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
The United States ◽  
Most Probable Number ◽  
Asian Countries ◽  
Probable Number ◽  
Specific Pcr ◽  
Thermostable Direct Hemolysin ◽  
Chromogenic Agar ◽  
Pcr Method ◽  
Arbitrarily Primed Pcr

ABSTRACT Although thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus has caused many infections in Asian countries, the United States, and other countries, it has been difficult to detect the same pathogen in seafoods and other environmental samples. In this study, we detected and enumerated tdh gene-positive V. parahaemolyticus in Japanese seafoods with a tdh-specific PCR method, a chromogenic agar medium, and a most-probable-number method. The tdh gene was detected in 33 of 329 seafood samples (10.0%). The number of tdh-positive V. parahaemolyticus ranged from <3 to 93/10 g. The incidence of tdh-positive V. parahaemolyticus tended to be high in samples contaminated with relatively high levels of total V. parahaemolyticus. TDH-producing strains of V. parahaemolyticus were isolated from 11 of 33 tdh-positive samples (short-necked clam, hen clam, and rock oyster). TDH-producing strains of V. parahaemolyticus were also isolated from the sediments of rivers near the coast in Japan. Representative strains of the seafood and sediment isolates were examined for the O:K serovar and by the PCR method specific to the pandemic clone and arbitrarily primed PCR and pulsed-field gel electrophoresis techniques. The results indicated that most O3:K6 tdh-positive strains belonged to the pandemic O3:K6 clone and suggested that serovariation took place in the Japanese environment.

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