The Effects of Storage Temperature on the Growth of Vibrio parahaemolyticus and Organoleptic Properties in Oysters

During harvesting and storage, microbial pathogens and natural spoilage flora may grow, negatively affecting the composition and texture of oysters and posing a potential health threat to susceptible consumers. A solution to these problems would mitigate associated damaging effects on the seafood industry. The purpose of this study was to investigate the effects of storage temperature on growth of Vibrios as well as other microbial, sensory and textural characteristics of post-harvest shellstock Eastern oysters (Crassostrea virginica). Oysters harvested from the Chesapeake Bay, Maryland, during summer months (June, July, and August, 2010) were subjected to three storage temperatures (5, 10, and 20ºC) over a 10 day period. At selected time intervals (0, 1, 3, 7, 10 days) two separate samples of 6 oysters each were homogenated and analyzed for pH, halophilic plate counts (HPC), total Vibrios, and Vibrio parahaemolyticus (Vp). Oyster meats shucked after storage were also organoleptically evaluated (acceptability, appearance, and odor). Texture analysis was performed using a texture analyzer on meats shucked from oysters held under the same conditions. The pH of the oyster homogenates showed no consistent pattern with storage time and temperature. The HPC (4.5 to 9.4log CFU/g) were highest on day 7 at 20C while olfactory acceptance reduced with time and increasing storage temperatures. The Vp counts increased over time from 3.5 to7.5 log MPN/g by day10. Loss of freshness as judged by appearance and odor was significant over time (p <br/

The Effect of Previous Life Cycle Phase on the Growth Kinetics, Morphology, and Antibiotic Resistance of Salmonella Typhimurium DT104 in Brain Heart Infusion and Ground Chicken Extract

Growth models are predominately used in the food industry to estimate the potential growth of selected microorganisms under environmental conditions. The growth kinetics, cellular morphology, and antibiotic resistance were studied throughout the life cycle of Salmonella Typhimurium. The effect of the previous life cycle phase [late log phase (LLP), early stationary phase (ESP), late stationary phase (LSP), and early death phase (EDP)] of Salmonella after reinoculation in brain heart infusion broth (BHI), ground chicken extract (GCE), and BHI at pH 5, 7, and 9 and salt concentrations 2, 3, and 4% was investigated. The growth media and previous life cycle phase had significant effects on the lag time (λ), specific growth rate (μmax), and maximum population density (Ymax). At 2 and 4% salt concentration, the LLP had the significantly (p &lt; 0.05) fastest μmax (1.07 and 0.69 log CFU/ml/h, respectively). As the cells transitioned from the late log phase (LLP) to the early death phase (EDP), the λ significantly (p &lt; 0.05) increased. At pH 5 and 9, the EDP had a significantly (p &lt; 0.05) lower Ymax than the LLP, ESP, and LSP. As the cells transitioned from a rod shape to a coccoid shape in the EDP, the cells were more susceptible to antibiotics. The cells regained their resistance as they transitioned back to a rod shape from the EDP to the log and stationary phase. Our results revealed that growth kinetics, cell’s length, shape, and antibiotic resistance were significantly affected by the previous life cycle phase. The results of this study also demonstrate that the previous life cycle should be considered when developing growth models of foodborne pathogens to better ensure the safety of poultry and poultry products

Harnessing Naturally Occurring Tumor Immunity: A Clinical Vaccine Trial in Prostate Cancer

International audienceBACKGROUND:Studies of patients with paraneoplastic neurologic disorders (PND) have revealed that apoptotic tumor serves as a potential potent trigger for the initiation of naturally occurring tumor immunity. The purpose of this study was to assess the feasibility, safety, and immunogenicity of an apoptotic tumor-autologous dendritic cell (DC) vaccine.METHODS AND FINDINGS:We have modeled PND tumor immunity in a clinical trial in which apoptotic allogeneic prostate tumor cells were used to generate an apoptotic tumor-autologous dendritic cell vaccine. Twenty-four prostate cancer patients were immunized in a Phase I, randomized, single-blind, placebo-controlled study to assess the safety and immunogenicity of this vaccine. Vaccinations were safe and well tolerated. Importantly, we also found that the vaccine was immunogenic, inducing delayed type hypersensitivity (DTH) responses and CD4+ and CD8+ T cell proliferation, with no effect on FoxP3+ regulatory T cells. A statistically significant increase in T cell proliferation responses to prostate tumor cells in vitro (p = 0.002), decrease in prostate specific antigen (PSA) slope (p = 0.016), and a two-fold increase in PSA doubling time (p = 0.003) were identified when we compared data before and after vaccination.CONCLUSIONS:An apoptotic cancer cell vaccine modeled on naturally occurring tumor immune responses in PND patients provides a safe and immunogenic tumor vaccine

Shifting the limits in wheat research and breeding using a fully annotated reference genome

Introduction: Wheat (Triticum aestivum L.) is the most widely cultivated crop on Earth, contributing about a fifth of the total calories consumed by humans. Consequently, wheat yields and production affect the global economy, and failed harvests can lead to social unrest. Breeders continuously strive to develop improved varieties by fine-tuning genetically complex yield and end-use quality parameters while maintaining stable yields and adapting the crop to regionally specific biotic and abiotic stresses. Rationale: Breeding efforts are limited by insufficient knowledge and understanding of wheat biology and the molecular basis of central agronomic traits. To meet the demands of human population growth, there is an urgent need for wheat research and breeding to accelerate genetic gain as well as to increase and protect wheat yield and quality traits. In other plant and animal species, access to a fully annotated and ordered genome sequence, including regulatory sequences and genome-diversity information, has promoted the development of systematic and more time-efficient approaches for the selection and understanding of important traits. Wheat has lagged behind, primarily owing to the challenges of assembling a genome that is more than five times as large as the human genome, polyploid, and complex, containing more than 85% repetitive DNA. To provide a foundation for improvement through molecular breeding, in 2005, the International Wheat Genome Sequencing Consortium set out to deliver a high-quality annotated reference genome sequence of bread wheat. Results: An annotated reference sequence representing the hexaploid bread wheat genome in the form of 21 chromosome-like sequence assemblies has now been delivered, giving access to 107,891 high-confidence genes, including their genomic context of regulatory sequences. This assembly enabled the discovery of tissue- and developmental stage–related gene coexpression networks using a transcriptome atlas representing all stages of wheat development. The dynamics of change in complex gene families involved in environmental adaptation and end-use quality were revealed at subgenome resolution and contextualized to known agronomic single-gene or quantitative trait loci. Aspects of the future value of the annotated assembly for molecular breeding and research were exemplarily illustrated by resolving the genetic basis of a quantitative trait locus conferring resistance to abiotic stress and insect damage as well as by serving as the basis for genome editing of the flowering-time trait. Conclusion: This annotated reference sequence of wheat is a resource that can now drive disruptive innovation in wheat improvement, as this community resource establishes the foundation for accelerating wheat research and application through improved understanding of wheat biology and genomics-assisted breeding. Importantly, the bioinformatics capacity developed for model-organism genomes will facilitate a better understanding of the wheat genome as a result of the high-quality chromosome-based genome assembly. By necessity, breeders work with the genome at the whole chromosome level, as each new cross involves the modification of genome-wide gene networks that control the expression of complex traits such as yield. With the annotated and ordered reference genome sequence in place, researchers and breeders can now easily access sequence-level information to precisely define the necessary changes in the genomes for breeding programs. This will be realized through the implementation of new DNA marker platforms and targeted breeding technologies, including genome editing

Development of a Simple and Rapid Fluorogenic Procedure for Identification of Vibrionaceae Family Members

We describe a simple colony overlay procedure for peptidases (COPP) for the rapid fluorogenic detection and quantification of Vibrionaceae from seawater, shellfish, sewage, and clinical samples. The assay detects phosphoglucose isomerase with a lysyl aminopeptidase activity that is produced by Vibrionaceae family members. Overnight cultures are overlaid for 10 min with membranes containing a synthetic substrate, and the membranes are examined for fluorescent foci under UV illumination. Fluorescent foci were produced by all the Vibrionaceae tested, including Vibrio spp., Aeromonas spp., and Plesiomonas spp. Fluorescence was not produced by non-Vibrionaceae pathogens. Vibrio cholerae strains O1, O139, O22, and O155 were strongly positive. Seawater and oysters were assayed, and 87 of 93 (93.5%) of the positive isolates were identified biochemically as Vibrionaceae, principally Vibrio vulnificus, Vibrio parahaemolyticus, Aeromonas hydrophila, Photobacterium damselae, and Shewanella putrefaciens. None of 50 nonfluorescent isolates were Vibrionaceae. No Vibrionaceae were detected in soil, and only A. hydrophila was detected in sewage. The COPP technique may be particularly valuable in environmental and food-testing laboratories and for monitoring water quality in the aquaculture industry

Table_2_Assesments of Vibrio parahaemolyticus and Vibrio vulnificus levels and microbial community compositions in blue crabs (Callinectes sapidus) and seawater harvested from the Maryland Coastal Bays.XLSX

IntroductionFluctuations in environmental physicochemical parameters can affect the diversity and prevalence of microbial communities, including vibrios, associated with aquatic species and their surrounding environments. This study aimed to investigate the population dynamics of two Vibrio species as well as the microbial community diversity of whole crab and seawater from the Maryland Coastal Bays (MCBs), using 16S rRNA sequencing.MethodsDuring this study, three crabs and 1 L of seawater were collected monthly from two sites for 3 months. Crab tissue was extracted and pooled for each site. Extracted crab tissue and seawater were analyzed for Vibrio parahaemolyticus and V. vulnificus using Most Probable Number (MPN) real-time PCR. For 16S rRNA microbiome analysis, three different DNA extraction kits were evaluated to extract microbial DNA from individual crabs. Also, 500 mL of each seawater sample was filtered for DNA extraction.ResultsResults indicated that sample types and sampling periods had a significant effect on the alpha diversity of the microbial community of crabs and seawater (p DiscussionResults from this study provide further insight into species diversity and phylogenetic compositions of blue crabs and seawater from the MCBs. These approaches will help in risk assessments that are essential in the overall advancement of public health.</p

Molecular characterization of antibiotic resistant Salmonella Typhimurium and Salmonella Kentucky isolated from pre-and post-chill whole broilers carcasses

a b s t r a c t There is conflicting data regarding whether commercial chilling has any effect on persistence of Salmonella serovars, including antibiotic resistant variants, on chicken carcasses. A total of 309 Salmonella Typhimurium and Salmonella Kentucky isolates recovered from pre-and post-chill whole broiler carcasses were characterized for genetic relatedness using Pulsed Field Gel Electrophoresis (PFGE) and for the presence of virulence factors (invA, pagC, spvC) by PCR and for aerobactin and colicin production by bioassays. A subset of these isolates (n ¼ 218) displaying resistance to either sulfisoxazole and/or ceftiofur [S. Typhimurium (n ¼ 66) and S. Kentucky (n ¼ 152)] were further tested for the presence of associated antibiotic resistance elements (class-I integrons and bla CMY genes) by PCR. All 145 ceftiofur resistant S. Kentucky and S. Typhimurium isolates possessed bla CMY genes. Class-I integrons were only detected in 6.1% (n ¼ 4/66) of sulfisoxazole resistant S. Typhimurium isolates. The PFGE analysis revealed the presence of genetically diverse populations within the recovered isolates but clusters were generally concordant with serotypes and antimicrobial resistance profiles. At a 100% pattern similarity index, thirty-six percent of the undistinguishable S. Typhimurium and 22% of the undistinguishable S. Kentucky isolates were recovered from the same chilling step. All isolates possessed the invA and pagC genes, but only 1.4%possessed spvC. Irrespective of the chilling step, there was a significant difference (P &lt; 0.05) in the production of aerobactin and colicin between S. Typhimurium and S. Kentucky isolates. Taken together, these results indicate that chilling impacted the recovery of particular Salmonella clonal groups but had no effect on the presence of class-I integrons, bla CMY genes, and tested virulence factors

Molecular characterization of antibiotic resistant \u3ci\u3eSalmonella\u3c/i\u3e Typhimurium and \u3ci\u3eSalmonella\u3c/i\u3e Kentucky isolated from pre- and post-chill whole broilers carcasses

There is conflicting data regarding whether commercial chilling has any effect on persistence of Salmonella serovars, including antibiotic resistant variants, on chicken carcasses. A total of 309 Salmonella Typhimurium and Salmonella Kentucky isolates recovered from pre- and post-chill whole broiler carcasses were characterized for genetic relatedness using Pulsed Field Gel Electrophoresis (PFGE) and for the presence of virulence factors (invA, pagC, spvC) by PCR and for aerobactin and colicin production by bioassays. A subset of these isolates (n = 218) displaying resistance to either sulfisoxazole and/or ceftiofur [S. Typhimurium (n = 66) and S. Kentucky (n = 152)] were further tested for the presence of associated antibiotic resistance elements (class-I integrons and blaCMY genes) by PCR. All 145 ceftiofur resistant S. Kentucky and S. Typhimurium isolates possessed blaCMY genes. Class-I integrons were only detected in 6.1% (n = 4/66) of sulfisoxazole resistant S. Typhimurium isolates. The PFGE analysis revealed the presence of genetically diverse populations within the recovered isolates but clusters were generally concordant with serotypes and antimicrobial resistance profiles. At a 100% pattern similarity index, thirty-six percent of the undistinguishable S. Typhimurium and 22% of the undistinguishable S. Kentucky isolates were recovered from the same chilling step. All isolates possessed the invA and pagC genes, but only 1.4%possessed spvC. Irrespective of the chilling step, there was a significant difference (P \u3c 0.05) in the production of aerobactin and colicin between S. Typhimurium and S. Kentucky isolates. Taken together, these results indicate that chilling impacted the recovery of particular Salmonella clonal groups but had no effect on the presence of class-I integrons, blaCMY genes, and tested virulence factors