Histochemical localization of N-acetylhexosamine-binding lectin HOL-18 in Halichondria okadai (Japanese black sponge), and its antimicrobial and cytotoxic anticancer effects.

We studied localization and physiological activities of a lectin showing specific binding to N-acetylhexosamines, termed HOL-18, purified from Japanese black sponge (Halichondria okadai). Antiserum against the lectin was generated in rabbit and applied for immunohistochemical analyses. HOL-18 was expressed specifically around water pores and on spicules of sponge tissues. It showed strong binding to a variety of N-acetylhexosamines: N-acetyl D-glucosamine, N-acetyl D-galactosamine, N-acetyl mannosamine, N-acetyl muramic acid, and N-acetyl neuraminic acid. Hemagglutination induced by the lectin was inhibited by lipopolysaccharides and a peptidoglycan. HOL-18 inhibited growth of a gram-positive bacterium (Listeria monocytogenes), gram-negative bacteria (Escherichia coli, Shigella boydii, Pseudomonas aeruginosa), and a fungus (Aspergillus niger). It displayed anti-biofilm activity against P. aeruginosa. HOL-18 was internalized into conidiophores of A. niger, and displayed notable antifungal activity. Fluorescence microscopy revealed binding and incorporation of the lectin into human cancer cell lines HeLa, MCF-7, and T47D, but not Caco-2. HOL-18 displayed dose-dependent cytotoxic effects against HeLa, MCF-7, and T47D, with respective IC50 values 40, 52, and 63 µg/mL. In HeLa cells, it activated phosphorylation of MAPK pathway molecule (ERK1/2) and activated caspase-3 to trigger apoptosis. HOL-18 thus has the potential to upregulate metabolic pathways in higher animal cells through binding to N-acetylhexosamines.

A Shigella boydii bacteriophage which resembles Salmonella phage ViI.

BACKGROUND: Lytic bacteriophages have been applied successfully to control the growth of various foodborne pathogens. Sequencing of their genomes is considered as an important preliminary step to ensure their safety prior to food applications. RESULTS: The lytic bacteriophage, ΦSboM-AG3, targets the important foodborne pathogen, Shigella. It is morphologically similar to phage ViI of Salmonella enterica serovar Typhi and a series of phages of Acinetobacter calcoaceticus and Rhizobium meliloti. The complete genome of ΦSboM-AG3 was determined to be 158 kb and was terminally redundant and circularly permuted. Two hundred and sixteen open reading frames (ORFs) were identified and annotated, most of which displayed homology to proteins of Salmonella phage ViI. The genome also included four genes specifying tRNAs. CONCLUSIONS: This is the first time that a Vi-specific phage for Shigella has been described. There is no evidence for the presence of virulence and lysogeny-associated genes. In conclusion, the genome analysis of ΦSboM-AG3 indicates that this phage can be safely used for biocontrol purposes.

Arginine-dependent acid-resistance pathway in Shigella boydii.

Ability to survive the low pH of the human stomach is considered be an important virulent determinant. It was suggested that the unique acid tolerance of Shigella boydii 18 CDPH, the strain implicated in a 1998 outbreak, may have played an important role in surviving the acidic food (bean salad). The strain was capable of inducing arginine-dependent acid-resistance (ADAR) pathway. This pathway was assumed to be absent in Shigella sp. Here, we have examined occurrence and efficacy of ADAR pathway in 21 S. boydii strains obtained from the American Type Culture Collection (ATCC) along with strains of S. flexneri (n = 7), S. sonnei (n = 4), and S. dysenteriae (n = 2). The eight S. boydii strains were able to induce ADAR to survive the acid challenge at pH 2.0; additional 8 strains could tolerate acid challenge at pH 2.5 but not at pH 2.0. The remaining five S. boydii strains were not able to induce ADAR pathway and could not survive acid challenge even at pH 2.5. ADAR pathway also appears to be present in all four Shigella sp. Shigella ADAR pathway was induced when cells were grown under partial oxygen pressure while its expression in E. coli required mere fermentative growth on glucose.

Effect of NaCl on the biofilm formation by foodborne pathogens.

This study was designed to evaluate the effect of NaCl on the biofilm formation of Listeria monocytogenes, Staphylococcus aureus, Shigella boydii, and Salmonella Typhimurium. The biofilm cells were cultured in media containing different NaCl concentrations (0% to 10%) for 10 d of incubation at 37 °C using a 24-well polystyrene microtiter plate, collected by swabbing methods, and enumerated using plate count method. The attachment and detachment kinetic patterns were estimated according to the modified Gompertz model. The cell surface hydrophobicity and auto-aggregation were observed at different NaCl concentrations. Most strains showed 2 distinctive phases at lower than 6% NaCl, while the numbers of adhered cells gradually increased throughout the incubation period at 4% to 10% NaCl. At 0% NaCl, the numbers of adhered L. monocytogenes, S. aureus, S. boydii, and S. Typhimurium cells rapidly increased up to 7.04, 6.47, 6.39, and 7.27 log CFU/cm(2), respectively, within 4 d of incubation. The maximum growth rate (k(A)) and specific growth rate (µ(A)) of adherent pathogenic cells were decreased with increasing NaCl concentration. Noticeable decline in the numbers of adherent cells was observed at low concentration levels of NaCl (<2%). The adherence abilities of foodborne pathogens were influenced by the physicochemical surface properties. The hydrophobicity and auto-aggregation enhanced the biofilm formation during the incubation periods. Therefore, this study could provide useful information to better understand the adhesion and detachment capability of foodborne pathogens on food contact surfaces.

Inactivation of Shigella boydii 18 IDPH and Listeria monocytogenes Scott A with power ultrasound at different acoustic energy densities and temperatures.

The effect of acoustic energy density (AED) on inactivation of Shigella boydii 18 IDPH and Listeria monocytogenes Scott A in a cell suspension was studied at sublethal temperatures and at AEDs of 0.49, 0.85, and 1.43 W/mL. The effect of temperature on ultrasonic inactivation of L. monocytogenes Scott A at 35, 50, and 65 degrees C was examined at an AED of 1.43 W/mL. Increasing AED increased the rate of inactivation for both S. boydii and L. monocytogenes. The destruction of S. boydii and L. monocytogenes followed 1st order kinetics in a 20-min treatment, except for S. boydii inactivation at 1.43 W/mL where a tailing effect was observed after 15 min. At sublethal temperatures, the D-values of S. boydii were 8.8, 4.3, and 2.5 min for AEDs of 0.49, 0.85, and 1.43 W/mL, whereas those for L. monocytogenes at the 3 AED levels were 31.5, 13.5, and 7.3 min, respectively. Ultrasonic treatment of L. monocytogenes at 35 and 50 degrees C enhanced inactivation. However, at 65 degrees C, application of ultrasound did not result in additional inactivation compared to thermal treatment alone at the same temperature. With the experimental conditions and the ultrasound system used in this study, an upper temperature limit for thermosonication was evident above which no added killing due to ultrasound was observed.

Survival of Shigella boydii 18 in bean salad.

A strain of Shigella boydii 18 involved in a bean salad outbreak and S. boydii 18 ATCC 35966 were used to inoculate bean salad. Bean salad samples were stored at 4 or 23 degrees C. At 4 degrees C, the S. boydii survived for the duration of the shelf life of the salad but did not grow. At 23 degrees C, both strains increased by two orders of magnitude by day 2 and decreased rapidly thereafter. This demonstrates the importance of proper storage in preventing the outgrowth of foodborne pathogens.

Comparison of chromogenic Shigella spp. plating medium with standard media for the recovery of Shigella boydii and Shigella sonnei from tomato surfaces.

Isolation of Shigella spp. from food is difficult because of a lack of appropriate selective media and the presence of low numbers of shigellae relative to competitive microorganisms. Chromogenic Shigella spp. plating medium (CSPM) was evaluated for use with the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) enrichment procedure for isolation of artificially contaminated Shigella boydii UI02 and Shigella sonnei UI05 from tomato surfaces. Tomatoes were inoculated with various concentrations of S. boydii UI02 or S. sonnei UI05 and rinsed using a shake-rub-shake procedure. Tomato rinses were enriched overnight according to the BAM procedure and streaked for isolation on CSPM, Salmonella-Shigella agar (SSA), and MacConkey agar (MAC). To access the isolation of S. boydii UI02 and S. sonnei UI05 without competition from natural tomato microflora, experiments were repeated using rifampin-adapted inocula and enrichments supplemented with 50 microg/ml rifampin. Isolation of S. boydii UI02 and S. sonnei UI05 with or without natural tomato microflora was not significantly different (P > 0.05) on CSPM, MAC, or SSA. Colony color enhancements created by CSPM may ease differentiation of Shigella colonies from those of closely related competitors.

Catecholamines and in vitro growth of pathogenic bacteria: enhancement of growth varies greatly among bacterial species.

The purpose of this study was to examine the effects of catecholamines on in vitro growth of a range of bacterial species, including anaerobes. Bacteria tested included: Porphyromonas gingivalis, Bacteriodes fragilis, Shigella boydii, Shigella sonnie, Enterobacter Sp, and Salmonella choleraesuis. The results of the current study indicated that supplementation of bacterial cultures in minimal medium with norepinephrine or epinephrine did not result in increased growth of bacteria. Positive controls involving treatment of Escherichia coli with catecholamines did result in increased growth of that bacterial species. The results of the present study extend previous observations that showed differential capability of catecholamines to enhance bacterial growth in vitro.

Shigella em alimentos / Shigella in the foods

Comenta-se nesta breve revisao a participacao e o significado de Shigella em processos de infeccao alimentar. Sao tambem abordados as caracteristicas do microrganismo, seus fatores de virulencia e determinancia genetica.Aspectos epidemiologicos da infeccao, bem como, condicoes de crescimento e metodos de deteccao em alimentos, sao tambem abordados

Bactericidal activities of selected organic N-halamines.

The bactericidal efficacies of three organic N,N'-dihalamine disinfectants in the class of compounds termed imidazolidinones were determined for combinations of pH, temperature, and water quality treatments by using Staphylococcus aureus and Shigella boydii as test organisms. The compound 1,3-dibromo-4,4,5,5-tetramethyl-2-imidazolidinone was found to be the most rapidly acting bactericide, especially under halogen-demand-free conditions. The mixed N,N'-dihalamine 1-bromo-3-chloro-4,4,5,5-tetramethyl-2-imidazolidinone was found to be intermediate in terms of rate of disinfection, while the compound 1,3-dichloro-4,4,5,5-tetramethyl-2-imidazolidinone was observed to be the slowest acting bactericide. When overall effectiveness was judged on the basis of stability of the disinfectants along with rates of disinfection, the mixed halamine was considered to exhibit great potential for use as a disinfectant in an aqueous solution.

Bacterial culture longevity: control by inorganic phosphate and temperature.

Death of cells in cultures of six species of gram-negative bacteria was accelerated by incorporation in the medium of 10-fold more inorganic phosphate than was needed for growth. Death induced by inorganic phosphate was independent of the carbohydrate employed and was enhanced by warm temperatures.