BackgroundANGUSTIFOLIA (AN), one of the CtBP family proteins, plays a major role in microtubule-dependent cell morphogenesis. Microarray analysis of mammalian AN homologs suggests that AN might function as a transcriptional activator and regulator of a wide range of genes. Genetic characterization of AN mutants suggests that AN might be involved in multiple biological processes beyond cell morphology regulation.ResultsUsing a reverse genetic approach, we provide in this paper the genetic, biochemical, and physiological evidence for ANGUSTIFOLIA’s role in other new biological functions such as abiotic and biotic stress response in higher plants. The T-DNA knockout an-t1 mutant exhibits not only all the phenotypes of previously described angustifolia null mutants, but also copes better than wild type under dehydration and pathogen attack. The stress tolerance is accompanied by a steady-state modulation of cellular H2O2 content, malondialdehyde (MDA) derived from cellular lipid peroxidation, and over-expression of stress responsive genes. Our results indicate that ANGUSTIFOLIA functions beyond cell morphology control through direct or indirect functional protein interaction networks mediating other biological processes such as drought and pathogen attacks.ConclusionsOur results indicate that the ANGUSTIFOLIA gene participates in several biochemical pathways controlling cell morphogenesis, abiotic, and biotic stress responses in higher plants. Our results suggest that the in vivo function of plant ANGUSTIFOLIA has been overlooked and it needs to be further studied beyond microtubule-dependent cell morphogenesis.
Aims: Clostridium perfringens type A causes both clinical and subclinical forms of necrotic enteritis in domestic avian species. In this study the inhibitory effect of hen egg white lysozyme on the vegetative form of Cl. perfringens type A and the production of α‐toxin in vitro was investigated.
Methods and Results: A micro‐broth dilution assay was used to evaluate the minimal inhibitory concentrations (MIC) of lysozyme against three clinical isolates of Cl. perfringens type A in 96‐well microtitre plates. The MIC of lysozyme against Cl. perfringens isolates was found to be 156 μg ml−1. Scanning electron micrographs of the cells treated with 100 μg ml−1 of lysozyme revealed extensive cell wall damage. A quantitative sandwich ELISA for α‐toxin produced by Cl. perfringens was developed based on a commercial ELISA kit allowing only qualitative detection. Addition of 50 μg ml−1 of lysozyme did not inhibit the growth of Cl. perfringens but significantly inhibited the toxin production.
Conclusions: Lysozyme inhibited the growth of Cl. perfringens type A at 156 μg ml−1. At sublethal levels, lysozyme was able to inhibit the α‐toxin production.
Significance and Impact of Study: Inhibition of Cl. perfringens type A and its α‐toxin production by hen egg white lysozyme had never previously been reported. By inhibiting this avian pathogen and its toxin production, lysozyme showed potential for use in the treatment and prevention of necrotic enteritis and other Cl. perfringens type A related animal diseases.
Many microorganisms (including a number of important foodborne pathogens) can be present on raw fruits and vegetables. Since these products are frequently eaten raw, any pathogens present represent a potential risk to the consumer. The objective of this study was to compare the efficacy of a commercial produce wash with that of water for reducing the total bacterial population on lettuce when used by food service employees in university dining halls. Because this study was carried out in actual food service facilities during their daily operation, we used indigenous produce microflora instead of actual pathogens. Over the course of the study, more than 40 heads of lettuce were divided into thirds, and each section was analyzed for total plate count either before washing, after washing in water, or after washing in Victory produce wash. When initial contamination levels were > or = 100 CFU/g (n = 36 samples), reductions obtained with Victory produce wash (1.8 log CFU/g) were significantly larger (P = 0.0006) than those obtained with water (0.8 log CFU/g). Our results indicate that Victory produce wash is effective in reducing indigenous flora on lettuce during food service preparation. Our results also show that care must be taken in the analysis of microbial reduction data: only a slight reduction in total plate count (ca. 0.1 log CFU/g) and no significant difference in reductions (P = 0.84) were observed when all samples (irrespective of initial contamination level) were compared.
Proper temperature control is essential in preventing Clostridium perfringens food poisoning. The U.S. Department of Agriculture Food Safety and Inspection Service cooling guidelines offer two options for the cooling of meat products: follow a standard time-temperature schedule or validate that alternative cooling regimens result in no more than a 1-log CFU/g increase of C. perfringens and no growth of Clostridium botulinum. The latter option requires laboratory challenge studies to validate the efficacy of a given cooling process. Accordingly, the objective of this study was to investigate the role of several methodological variables that might be encountered during typical C. perfringens challenge studies. Variables studied included plastic bag type (Whirlpak or Spiral Biotech), sealing method (Multivac or FoodSaver), initial spore inoculum size (1 to approximately 3 log CFU/g), and growth environment (ground beef or Trypticase-peptone-glucose-yeast extract [TPGY] broth). The major factors that affected growth were sample bag type and growth environment. Samples incubated in Whirlpak bags showed significantly less growth than those incubated in Spiral Biotech bags, which was likely due to the former bag's greater oxygen permeability. C. perfringens spores showed shorter germination, outgrowth, and lag times and C. perfringens cells showed faster growth rates in ground beef compared with TPGY broth. No significant difference was observed between two different sealing methods. Initial spore inoculum levels in the range studied had no significant effect on final C. perfringens cell concentration.
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