Staphylococcus aureus Growth and Enterotoxin A Production in an Anaerobic Environment

2002 ◽  
Vol 65 (1) ◽  
pp. 199-204 ◽  
Author(s):  
NEGASH BELAY ◽  
AVRAHAM RASOOLY
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Density ◽  
Incubation Temperature ◽  
Food Poisoning ◽  
Brain Heart Infusion ◽  
Toxin Production ◽  
Anaerobic Growth ◽  
Anaerobic Conditions ◽  
Effects Of Temperature ◽  
Anaerobic Environment

The effects of strict anaerobic conditions on the growth of Staphylococcus aureus and the production of staphylococcal enterotoxin A (SEA) were studied. The growth of S. aureus, a facultative anaerobic bacterium, is slower anaerobically than aerobically. When grown on brain heart infusion broth at 37°C, the anaerobic generation time at mid-log phase was 80 min, compared with 35 min for the aerobic control. In contrast to previous studies demonstrating that staphylococcal cell density was 9- to 17-fold greater in aerobic than in anaerobic cultures, data for a staphylococcal strain implicated in food poisoning showed that the cell density was only two to three times as great in aerobic cultures. Production of SEA was monitored by Western immunoblotting and shown to be growth dependent. With slower anaerobic growth, relatively less toxin was produced than under aerobic conditions, but in both cases SEA was detected after 120 min of incubation. The combined effects of temperature and aeration on S. aureus were also studied. Growth and toxin production of aerobic and anaerobic cultures at temperatures ranging from 14 to 37°C were analyzed. Growth was still observed at low temperatures in both environments. A linear model for S. aureus aerobic or anaerobic growth as a function of incubation temperature was developed from these studies. The model was tested from 17 to 35.5°C, and the results suggest that the model can accurately predict the S. aureus growth rate in this temperature range. The data suggest that anaerobic conditions are not an effective barrier against S. aureus growth.

scholarly journals Anaerobic Conditions Induce Expression of Polysaccharide Intercellular Adhesin in Staphylococcus aureus and Staphylococcus epidermidis

2001 ◽  
Vol 69 (6) ◽  
pp. 4079-4085 ◽  
Author(s):  
Sarah E. Cramton ◽  
Martina Ulrich ◽  
Friedrich Götz ◽  
Gerd Döring
Keyword(s):  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Extracellular Polysaccharide ◽  
Growth Conditions ◽  
Anaerobic Conditions ◽  
Anaerobic Environment ◽  
Specific Mrna

ABSTRACT Products of the intercellular adhesion (ica) operon in Staphylococcus aureus and Staphylococcus epidermidis synthesize a linear β-1,6-linked glucosaminylglycan. This extracellular polysaccharide mediates bacterial cell-cell adhesion and is required for biofilm formation, which is thought to increase the virulence of both pathogens in association with prosthetic biomedical implants. The environmental signal(s) that triggers ica gene product and polysaccharide expression is unknown. Here we demonstrate that anaerobic in vitro growth conditions lead to increased polysaccharide expression in both S. aureus and S. epidermidis, although the regulation is less stringent inS. epidermidis. Anaerobiosis also dramatically stimulates ica-specific mRNA expression inica- and polysaccharide-positive strains of both S. aureus and S. epidermidis.These data suggest a mechanism whereby ica gene expression and polysaccharide production may act as a virulence factor in an anaerobic environment in vivo.

Characteristics of Enterotoxin H–Producing Staphylococcus aureus Isolated from Clinical Cases and Properties of the Enterotoxin Productivity

2008 ◽  
Vol 71 (9) ◽  
pp. 1855-1860 ◽  
Author(s):  
FUMIHIKO SAKAI ◽  
HIDESHI IHARA ◽  
KENJI AOYAMA ◽  
HIDEO IGARASHI ◽  
SHUICHI YANAHIRA ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Water Activity ◽  
Food Poisoning ◽  
Brain Heart Infusion ◽  
Enzyme Linked Immunosorbent Assay ◽  
Late Exponential Phase ◽  
Epidemiological Characteristic ◽  
Predominant Type ◽  
High Prevalence ◽  
The Relationship

Staphylococcal enterotoxin H (SEH) is predicted to be involved in staphylococcal food poisoning. To characterize SEH-producing Staphylococcus aureus isolates from staphylococcal food poisoning cases in Japan, we investigated the relationship between SEH production and coagulase serotype, which is an epidemiological marker, and compared the properties of SEH production with those of staphylococcal enterotoxins A (SEA) and B (SEB). SEH production was determined by a newly developed sandwich enzyme-linked immunosorbent assay. Eighty-six (59.7%) of 144 isolates from staphylococcal food poisoning cases produced SEH. Seventy-one of the SEH-producing isolates simultaneously produced SEA, SEB, or both. All SEH-producing isolates belonged to coagulase type VII, which was the predominant type, representing 99 (68.8%) of 144 isolates. The amount of SEH produced in brain heart infusion was almost the same as the amount of SEA and approximately 10-fold lower than that of SEB. SEH and SEA were produced mainly during the late exponential phase of growth, whereas SEB was produced mostly during the stationary phase. The production levels of SEH and SEA were gradually affected by decreases in water activity, but the production of SEB was greatly reduced under conditions of low water activity. These findings indicate that SEH-producing S. aureus isolates are of high prevalence in staphylococcal food poisoning cases. Given the unique epidemiological characteristic of these isolates, SEH and SEA probably are responsible for food poisoning.

scholarly journals Growth characterisation of Staphylococcus aureus in milk: a quantitative approach

2009 ◽  
Vol 27 (No. 6) ◽  
pp. 433-453 ◽  
Author(s):  
A. Medveďová ◽  
Ľ. Valík ◽  
Z. Sirotná ◽  
D. Liptáková
Keyword(s):  
Staphylococcus Aureus ◽  
Growth Rate ◽  
Incubation Temperature ◽  
Food Poisoning ◽  
Raw Milk ◽  
Lag Phase ◽  
Phase Duration ◽  
Temperature Range ◽  
Root Model ◽  
Toxigenic Strains

<i>Staphylococcus aureus</i> is a pathogenic bacterium that induces several of human illnesses. The staphylococcal enterotoxin (SE) production as the results of previous growth of toxigenic strains is the most crucial problem which may lead to the staphylococcal food poisoning outbreaks in humans. That is why the growth of three strains of <i>Staphylococcus aureus</i> was characterised in milk and modelled in dependence of temperature. For the lag phase duration of <i>S. aureus</i> 2064, the Davey model was used with the following result: ln(1/lag) = 1.973 – 87.92/<i>T</i> + 285.09/<i>T</i><sup>2</sup> (<i>R</i><sup>2</sup> = 0.962). The dependence of the growth rate on incubation temperature was modelled by the Ratkowsky square root model and Gibson in sub-optimal and whole temperature range, respectively. The validation of both models showed high significance of the growth rate data fitting. The optimal temperature of <i>T</i><sub>opt</sub> = 38.5°C was resulted from Gibson model for the <i>S. aureus</i> 2064 growth in milk. For practical purpose, the time necessary for the increase of <i>S. aureus</i> by 3 log counts was also calculated within the growth temperature range. These data may provide useful information e.g. for the producers using raw milk in their artisanal cheese practice as the specific strains were used in this study.

scholarly journals TSST-1, enterotoxin and bacteriocin-like substance production by Staphylococcus aureus isolated from foods

2013 ◽  
Vol 65 (5) ◽  
pp. 1537-1544 ◽  
Author(s):  
S.A. Carvalho ◽  
L.S. Carmo ◽  
E.F. Abreu ◽  
R.S. Dias ◽  
A.C.M. Apolônio ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Milk ◽  
Food Poisoning ◽  
Brain Heart Infusion ◽  
Food Samples ◽  
Antagonistic Activity ◽  
Toxic Shock ◽  
Toxic Shock Syndrome Toxin ◽  
Reference Strains ◽  
Infusion Agar

The production of Toxic Shock Syndrome Toxin-1 (TSST-1), enterotoxins and bacteriocin-like substances was evaluated in 95 strains of Staphylococcus aureus recovered from raw bovine milk (n=31) and from food samples involved in staphylococcal food poisoning (n=64). Enterotoxigenicity tests with the membrane over agar associated to optimal sensibility plate assays were performed and showed that 96.77% of strains recovered from milk and 95.31% from food samples produced enterotoxins A, B, C, D or TSST-1. Reference strains S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella Typhimurium, Escherichia coli, Enterococcus faecalis and Bacteroides fragilis were used as indicator bacteria in the antagonistic assays, the first five being sensitive to antagonistic substances. Brain heart infusion agar, in pH values ranging from 5.0 to 7.0 in aerobic atmosphere showed to be the optimum condition for antagonistic activity as evaluated with the best producer strains against the most sensitive indicator bacterium, L. monocytogenes. Sensitivity to enzymes confirmed the proteinaceous nature of these substances. Neither bacteriophage activity nor fatty acids were detected and the antagonistic activity was not due to residual chloroform. Results did not establish a positive correlation between the bacteriocinogenic profile and toxigenicity in the tested S. aureus strains.

Modeling the Growth Boundary of Staphylococcus aureus for Risk Assessment Purposes

2001 ◽  
Vol 64 (1) ◽  
pp. 51-57 ◽  
Author(s):  
CYNTHIA M. STEWART ◽  
MARTIN B. COLE ◽  
J. DAVID LEGAN ◽  
LOUISE SLADE ◽  
MARK H. VANDEVEN ◽  
...  
Keyword(s):  
Risk Assessment ◽  
Staphylococcus Aureus ◽  
Brain Heart Infusion ◽  
Toxin Production ◽  
Potassium Sorbate ◽  
Growth Responses ◽  
Microtiter Plates ◽  
Initial Ph ◽  
Data Points ◽  
Boundary Model

Knowing the precise boundary for growth of Staphylococcus aureus is critical for food safety risk assessment, especially in the formulation of safe, shelf-stable foods with intermediate relative humidity (RH) values. To date, most studies and resulting models have led to the presumption that S. aureus is osmotolerant. However, most studies and resulting models have focused on growth kinetics using NaCl as the humectant. In this study, glycerol was used to investigate the effects of a glass-forming nonionic humectant to avoid specific metabolic aspects of membrane ion transport. The experiments were designed to produce a growth boundary model as a tool for risk assessment. The statistical effects and interactions of RH (84 to 95% adjusted by glycerol), initial pH (4.5 to 7.0 adjusted by HCl), and potassium sorbate (0, 500, or 1,000 ppm) or calcium propionate (0, 500, or 1,000 ppm) on the aerobic growth of a five-strain S. aureus cocktail in brain heart infusion broth were explored. Inoculated broths were distributed into microtiter plates and incubated at 37°C over appropriate saturated salt slurries to maintain RH. Growth was monitored by turbidity during a 24-week period. Toxin production was explored by enterotoxin assay. The 1,280 generated data points were analyzed by SAS LIFEREG procedures, which showed all studied parameters significantly affected the growth responses of S. aureus with interactions between RH and pH. The resulting growth/no growth boundary is presented.

Growth and Enterotoxin Production of Staphylococcus aureus in Fresh Packaged Mushrooms (Agaricus bisporus)

1996 ◽  
Vol 59 (8) ◽  
pp. 819-826 ◽  
Author(s):  
STEFAN T. MARTIN ◽  
ROBERT B. BEELMAN
Keyword(s):  
Staphylococcus Aureus ◽  
Food Chain ◽  
Agaricus Bisporus ◽  
Incubation Temperature ◽  
Staphylococcal Enterotoxin ◽  
Low Level ◽  
Enterotoxin Production ◽  
Different Temperatures ◽  
Storage Temperatures ◽  
Anaerobic Environment

Freshly harvested mushrooms were found to induce a near-anaerobic environment (&lt;2% O2) in unventilated, PVC-overwrapped packages within 2 to 6 h when incubated at 20 to 30°C. Mushrooms were inoculated with an enterotoxigenic strain of Staphylococcus aureus and incubated in overwrapped trays at different temperatures. S. aureus grew and produced staphylococcal enterotoxin (SE) in unventilated PVC-overwrapped mushroom packages when inoculated at levels of 103, 104, and 105 CFU/g of mushroom after 4 days of incubation at 30°C. Growth of S. aureus was observed at all levels of inoculation at 25°C, but no SE was detected after 7 days of incubation. When mushroom packages were ventilated, S. aureus growth was suppressed and no SE was detected after 7 days at 25°C and 4 days at 30°C. However, S. aureus growth in ventilated packs exceeded growth in unventilated packages when the incubation temperature was increased to 35°C; SE was detected within 18 h of incubation at this temperature, even in mushrooms inoculated at a low level (102 CFU/g). These results show the extreme importance of proper sanitation and worker hygiene during mushroom harvesting and packaging, ventilation of fresh mushroom packages, and proper storage temperatures for fresh mushrooms at all points of the food chain.

Effect of Borate on the Growth of Coagulase-Positive and Coagulase-Negative Staphylococci

1970 ◽  
Vol 1 (1) ◽  
pp. 78-79
Author(s):  
James M. Jay
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Toxin Production ◽  
Sodium Borate ◽  
Coagulase Negative Staphylococci

Of 235 strains of coagulase-positive Staphylococcus aureus studied, 221 or 94% were inhibited by 1.5 × 10 −8 m sodium borate, whereas only 6 of 57 (10.5%) coagulase-negative strains were inhibited by the same borate concentration. Four of the six coagulase-negatives were food poisoning strains and one was a hospital pathogen. Borate sensitivity was found to correlate well with lysozyme and α-toxin production by coagulase-positive strains.

scholarly journals Staphylococcus aureus ArcR Controls Expression of the Arginine Deiminase Operon

2007 ◽  
Vol 189 (16) ◽  
pp. 5976-5986 ◽  
Author(s):  
Julia Makhlin ◽  
Tzili Kofman ◽  
Ilya Borovok ◽  
Christian Kohler ◽  
Susanne Engelmann ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bioinformatic Analysis ◽  
Arginine Deiminase ◽  
Anaerobic Growth ◽  
Binding Motif ◽  
Sequence Motif ◽  
Sequence Motifs ◽  
Anaerobic Conditions ◽  
Catabolic Repression ◽  
Regulatory Regions

ABSTRACT We identified a single open reading frame that is strongly similar to ArcR, a member of the Crp/Fnr family of bacterial transcriptional regulators, in all sequenced Staphylococcus aureus genomes. The arcR gene encoding ArcR forms an operon with the arginine deiminase (ADI) pathway genes arcABDC that enable the utilization of arginine as a source of energy for growth under anaerobic conditions. In this report, we show that under anaerobic conditions, S. aureus growth is subject to glucose catabolic repression and is enhanced by arginine. Likewise, glucose and arginine have reciprocal effects on the transcription of the arcABDCR genes. Furthermore, we show using a mutant deleted for arcR that the transcription of the arc operon under anaerobic conditions depends strictly on a functional ArcR. These findings are supported by proteome analyses, which showed that under anaerobic conditions the expression of the ADI catabolic proteins depends on ArcR. Bioinformatic analysis of S. aureus ArcR predicts an N-terminal nucleotide binding domain and a C-terminal helix-turn-helix DNA binding motif. ArcR binds to a conserved Crp-like sequence motif, TGTGA-N6-TCACA, present in the arc promoter region and thereby activates the expression of the ADI pathway genes. Crp-like sequence motifs were also found in the regulatory regions of some 30 other S. aureus genes mostly encoding anaerobic enzymatic systems, virulence factors, and regulatory systems. ArcR was tested and found to bind to the regulatory regions of four such genes, adh1, lctE, srrAB, and lukM. In one case, for lctE, encoding l-lactate dehydrogenase, ArcR was able to bind only in the presence of cyclic AMP. These observations suggest that ArcR is likely to play an important role in the expression of numerous genes required for anaerobic growth.

scholarly journals Oxygen and Carbon Dioxide Regulation of Toxic Shock Syndrome Toxin 1 Production by Staphylococcus aureus MN8

2000 ◽  
Vol 38 (5) ◽  
pp. 1797-1803 ◽  
Author(s):  
Jeremy M. Yarwood ◽  
Patrick M. Schlievert
Keyword(s):  
Thin Film ◽  
Carbon Dioxide ◽  
Staphylococcus Aureus ◽  
Toxic Shock Syndrome ◽  
Toxin Production ◽  
Toxic Shock ◽  
Anaerobic Conditions ◽  
Batch Cultures ◽  
Toxic Shock Syndrome Toxin ◽  
Oxygen Concentrations

The production of toxic shock syndrome toxin 1 (TSST-1) byStaphylococcus aureus MN8 exposed to a range of oxygen concentrations (0 to 21% [vol/vol]) was examined in batch and thin-film cultures. The response of S. aureus to this range of oxygen concentrations was studied in the absence and in the presence of 7% (vol/vol) carbon dioxide. In the absence of carbon dioxide, TSST-1 production in batch cultures increased from negligible levels in the presence of oxygen concentrations of 1% or less to 500 ng/ml in the presence of 2% oxygen and then decreased to 70 ng/ml or less in the presence of oxygen concentrations of 6% and higher. In the presence of carbon dioxide, however, toxin production increased from negligible levels in the presence of 1% oxygen to 1,900 ng/ml in the presence of 21% oxygen. In thin-film cultures, TSST-1 production increased from nearly undetectable levels under anaerobic conditions to 1 and 10 μg/ml under 21% oxygen in the absence and presence of carbon dioxide, respectively. This study demonstrates the controlling effects of both oxygen and carbon dioxide on TSST-1 production.

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