Virulence factor genes encoding the thermostable direct hemolysin (tdh) and the thermostable direct hemolysin-related hemolysin (trh) are strongly correlated with virulence of the emergent human pathogen Vibrio parahaemolyticus. The gene encoding the thermolabile hemolysin (tlh) is also considered a signature molecular marker for the species. These genes are typically reported in very low percentages (1 to 2%) of nonclinical strains. V. parahaemolyticus strains were isolated from various niches within a pristine estuary (North Inlet, SC) and were screened for these genes using both newly designed PCR primers and more commonly used primers. DNA sequences of tdh and trh were recovered from 48% and 8.3%, respectively, of these North Inlet strains. The recovery of pathogenic V. parahaemolyticus strains in such high proportions from an estuarine ecosystem that is virtually free of anthropogenic influences indicates the potential for additional, perhaps environmental roles of the tdh and trh genes.

Using the memristive properties of vanadium dioxide, we experimentally demonstrate an adaptive filter by placing a memristor into an LC contour. This circuit reacts to the application of select frequency signals by sharpening the quality factor of its resonant response, and thus “learns” according to the input waveform. The proposed circuit employs only analog passive elements, and may find applications in biologically inspired processing and information storage. We also extend the learning-circuit framework mathematically to include memory-reactive elements, such as memcapacitors and meminductors, and show how this expands the functionality of adaptive memory filters.

Detection of the human pathogen Vibrio parahaemolyticus often relies on molecular biological analysis of species-specific virulence factor genes. These genes have been employed in determinations of V. parahaemolyticus population numbers and the prevalence of pathogenic V. parahaemolyticus strains. Strains of the Vibrionaceae species Photobacterium damselae, Vibrio diabolicus, Vibrio harveyi, and Vibrio natriegens, as well as strains similar to Vibrio tubiashii, were isolated from a pristine salt marsh estuary. These strains were examined for the V. parahaemolyticus hemolysin genes tdh, trh, and tlh and for the V. parahaemolyticus type III secretion system 2␣ gene vscC2 using established PCR primers and protocols. Virulence-related genes occurred at high frequencies in non-V. parahaemolyticus Vibrionaceae species. V. diabolicus was of particular interest, as several strains were recovered, and the large majority (>83%) contained virulence-related genes. It is clear that detection of these genes does not ensure correct identification of virulent V. parahaemolyticus. Further, the occurrence of V. parahaemolyticus-like virulence factors in other vibrios potentially complicates tracking of outbreaks of V. parahaemolyticus infections. Vibrio parahaemolyticus is the leading cause of seafood-associated gastroenteritis in the United States and the world (1). The U.S. Food and Drug Administration (1) estimates that 4,500 reported cases of V. parahaemolyticus gastroenteritis occur every year, and outbreaks of V. parahaemolyticus infections are increasing in frequency and expanding in geographic range (2, 3). This organism is ubiquitous in nearshore marine waters, and cell numbers are typically highest in surficial sediments (4) and in turbid waters bearing high loads of resuspended sediment (5, 6). Filterfeeding bivalve mollusks, such as oysters and mussels, can concentrate V. parahaemolyticus and other pathogenic vibrios (for examples, see references 7 and 8), resulting in levels in the mollusks capable of producing infection in a person that ingests them (9). Virulent V. parahaemolyticus strains are clearly a concern for seafood safety, and their detection is important anywhere that elevated levels of this organism are found.Detection of V. parahaemolyticus in shellfish and environmental samples is typically based on molecular biological analysis of specific genes, particularly genes exclusive to this species and those strongly correlated with pathogenicity. The gene encoding the thermolabile hemolysin (TLH), designated tlh, encodes a phospholipase A2 (10). While its contribution to V. parahaemolyticus pathogenicity is unknown, expression of this gene is upregulated under conditions mimicking the human intestine (11,12). tlh is considered to be a species-specific marker for V. parahaemolyticus (13,14) and is frequently employed to identify this species (1,13,(15)(16)(17)(18). Genes encoding the thermostable direct hemolysin (TDH) and the homologous thermostable direct hemolysin-related hemolysin (TRH), tdh and trh, respe...

Cell suspensions of Escherichia coli and Lactobacillus acidophilus were exposed to 600-ns pulsed electric fields (nsPEFs) at varying amplitudes or High-23.5 kV cm −1 ) and pulse numbers (0 (sham), 1, 5, 10, 100 or 1000) at a 1 hertz (Hz) repetition rate. The induced temperature rise generated at these exposure parameters, hereafter termed thermal gradient, was measured and applied independently to cell suspensions in order to differentiate inactivation triggered by electric field (E-field) from heating. Treated cell suspensions were plated and cellular inactivation was quantified by colony counts after a 24-hour (h) incubation period. Additionally, cells from both exposure conditions were incubated with various antibiotic-soaked discs to determine if nsPEF exposure would induce changes in antibiotic susceptibility. Results indicate that, for both species, the total delivered energy (amplitude, pulse number and pulse duration) determined the magnitude of cell inactivation. Specifically, for 18.5 and 23.5 kV cm −1 exposures, L. acidophilus was more sensitive to the inactivation effects of nsPEF than E. coli, however, for the 13.5 kV cm −1 exposures E. coli was more sensitive, suggesting that L. acidophilus may need to meet an E-field threshold before significant inactivation can occur. Results also indicate that antibiotic susceptibility was enhanced by multiple nsPEF exposures, as observed by increased zones of growth inhibition. Moreover, for both species, a temperature increase of ≤ 20 °C (89% of exposures) was not sufficient to significantly alter cell inactivation, whereas none of the thermal equivalent exposures were sufficient to change antibiotic susceptibility categories.

Vibrio parahaemolyticus is the leading cause of seafood-associated gastroenteritis and is most commonly transmitted by raw oysters. Consequently, detection of virulent strains of this organism in oysters is a primary concern for seafood safety. Vibrio parahaemolyticus levels were determined in 110 individual oysters harvested from two sampling sites in SC, USA. The majority of oysters (98%) contained low levels of presumptive V. parahaemolyticus However, two healthy oysters contained presumptive V. parahaemolyticus numbers that were unusually high. These two 'hot' oysters contained levels of presumptive V. parahaemolyticus within the gills that were ∼100-fold higher than the average for other oysters collected at the same date and location. Current V. parahaemolyticus detection practices require homogenizing a dozen oysters pooled together to determine V. parahaemolyticus numbers, a procedure that would dilute out V. parahaemolyticus in these 'hot' oysters. This study demonstrates the variability of V. parahaemolyticus densities taken from healthy, neighboring individual oysters in the environment. Additionally, environmental V parahaemolyticus isolates were screened for the virulence-related genes, tdh and trh, using improved polymerase chain reaction primers and protocols. We detected these genes, previously thought to be rare in environmental isolates, in approximately half of the oyster isolates.