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Staphylococcus aureus
Foodborne pathogen: Staphylococcal food-borne disease
Student name: Keerthana Kissanthomas
Student Id No. :A00280414
Introduction
• Gram positive bacteria (cocci shaped that
occur in clusters & member of Bacillota)
• Commensal bacteria in human microbiota.
• Opportunistic pathogen (common contributor
to skin diseases, sinusitis, and food poisoning)
Fig 1 – morphology of S. aureus under microscope
after gram staining (Ganguly, 2018)
Staphylococcal food-borne disease (SFP) results from the contamination of
food by S. aureus’s enterotoxins (SE). (Kadariya, Smith and Thapaliya, 2014)
Food intoxication (Staphyloenterotoxicosis)
• S. aureus bacteria is killed by cooking, but its toxins are resistant to heat
(withstand heat at 121°C for 10 min) and stomach enzymes
(proteases).
(M. Tallent, W. Bennett and M. Miller, 2022)
Symptoms and Incubation period
It is generally understood and accepted that S. aureus as a food
intoxicant presents a risk only when the growth to high numbers
occurs - >/= 100,000 CFU/g (>/= 1mg). (Rajkovic, 2016)
Primary symptoms: Vomiting, nausea, diarrhoea and
abdominal pain.
Secondary symptoms : dehydration, headache, myalgia and
fatigue
Incubation period: Symptoms generally occurs between 30
minutes to 8 hours (average of 3 hours) after ingestion.
Symptoms lasts 1-3 days. (CDC, 2018)
Images generated by (Bio render 2023
Food sources and transmissions
• Prefers to deposit their enterotoxins on poultry, meats, diary, bakery products and salads.
In raw foods, S. aureus does not compete effectively with indigenous bacteria. Source of contamination?
Air, dust, and food contact surfaces
can all act as carriers for S. aureus to
enter foods.
(Image by (WebstaurantStore, 2023)
.
Source: Argudín, Mendoza and Rodicio, 2010
Major source of contamination is poor
handling of cooked or processed foods
and followed by storage suitable.
(Image by blog.userve.com, 2022)
S. aureus is also found in dairy cattle
(cows), particularly those with
subclinical mastitis.
(Image by Webb, 2023)
Some Outbreaks
Table 1: some examples of outbreaks of Staphyloenterotoxicosis (Hennekinne, De Buyser and Dragacci, 2012)
Year Place, Country Food source Cases References
Pathogenesis/virulence & Mode of action (i)
Table 2: Main Staphylococcal enterotoxin (SE) of S. aureus involved in food (Pinchuk, Beswick and Reyes, 2010)
•After the consumption, enterotoxins are
absorbed in the abdomen and cause typical
gastroenteritis.
•Induces 5-hydroxytrytamine (5-HT) from mast
cells which stimulates the vomiting centre of the
brain
•Enterotoxins also cause damage to the
epithelial lining of the intestinal cells.
The genes for enterotoxin production are present in
pathogenicity islands in the chromosome (Arun K. Bhunia 2018)
Source: Dura, 2021
Fig 2: S. aureus enterotoxins in stomach
(Arun K. Bhunia 2018 citied in Dura, 2021)
Fig 3: S. aureus enterotoxins in intestinal
cells (Fisher E. L, et al. 2018)
Pathogenesis/virulence & Mode of action (ii)
• SEs are superantigens (SAgs) - stimulates many T-cells
• Enterotoxins form complex with MHC class II molecules on surface of antigen-
presenting cells
• Proliferates T-cells, producing a large amounts of inflammatory cytokines (IL-2, IFNγ,
IL-1, TNF-α).
• A high abundance of inflammatory cytokines stimulates the neuron receptors in the
intestinal tract and causes diarrhoea.
Source: Dura, 2021
Fig 4: SEs as superantigen (Fisher E. L, et al. 2018)
1. 15-year-old (healthy) boy presented with septic shock after eating meat from
a fast-food restaurant.
2. Developed food intoxication symptoms (diarrhoea) after 4 hours.
3. TSST-1 gene verified by real-time PCR analysis of the S.aureus strain.
Source: Goudsmit et al., 2021
• Food poising from S. aureus cause toxic shock syndrome, a SAgs called toxic
shock syndrome toxin 1 (TSST-1), is produced.
Detection
Source: Wu et al., 2016
Toxins not the bacterium itself
Prevention
• Prevent food from being held at an unsafe temperature
(between 4.4°C and 60°C) for more than 2 hours.
• Refrigerate as soon as possible (below 4 °C ).
• Food handlers practicing good hygiene.
• Avoid cross contamination (raw and cooked food).
• If reheating food, ensure the temperature reaches at least
74 °C .
Image by Evans, n.d shown in Pinterest n.d
References
• Argudín, M.Á., Mendoza, M.C. and Rodicio, M.R. (2010). Food Poisoning and Staphylococcus aureus Enterotoxins. Toxins, [online] 2(7), pp.1751–1773. doi:https://doi.org/10.3390/toxins2071751.
• blog.userve.com. (2022). Food Handler How-To: Handle Poultry Properly. [online] Available at: https://blog.userve.com/us/food-handler-how-to-handle-poultry-properly [Accessed 5 Nov. 2023].
• Centers for Disease Control and Prevention (2018). Staphylococcal (staph) food poisoning. [online] Centers for Disease Control and Prevention. Available at:
https://www.cdc.gov/foodsafety/diseases/staphylococcal.html.
• Ganguly, S. (2018). Photograph showing S. aureus grape cluster like morphology on Gram stain. 1000X. [Image] ResearchGate. Available at:
https://www.researchgate.net/publication/322243680_Molecular_Characterization_of_Staphylococcus_aureus_of_Camel_Camelus_dromedarius_Skin_Origin/figures?lo=1 [Accessed 30 Oct. 2023]
• Kadariya, J., Smith, T.C. and Thapaliya, D. (2014). Staphylococcus aureus and Staphylococcal food-borne disease: An ongoing challenge in public health. BioMed Research International, [online] 2014, pp.1–
9. doi:https://doi.org/10.1155/2014/827965
• Rajkovic, A. (2016). Staphylococcus: Food Poisoning. [online] , Ghent University, Ghent, Belgium & Belgrade University, Zemun-Belgrade, Serbia: Elsevier Ltd, pp.133–139. Available at:
https://elearning.unite.it/pluginfile.php/187875/mod_resource/content/0/rajkovic%2C2016-Staphylococcus-food%20poisoning.pdf [Accessed 2 Nov. 2023].
• Hennekinne, J.-A., De Buyser, M.-L. and Dragacci, S. (2012). Staphylococcus aureusand its food poisoning toxins: characterization and outbreak investigation. FEMS Microbiology Reviews, 36(4), pp.815–
836. doi:https://doi.org/10.1111/j.1574-6976.2011.00311.x.
• Pinchuk, I.V., Beswick, E.J. and Reyes, V.E. (2010). Staphylococcal Enterotoxins. Toxins, [online] 2(8), pp.2177–2197. doi:https://doi.org/10.3390/toxins2082177.
• Dura, S. (2021). Staphylococcal food poisoning (SFP)- Staphylococcus aureus enterotoxins. [online] Microbe Notes. Available at: https://microbenotes.com/staphylococcal-food-poisoning/ [Accessed 2 Nov.
2023].
• Fisher, E.L., Otto, M. and Cheung, G.Y.C. (2018). Basis of Virulence in Enterotoxin-Mediated Staphylococcal Food Poisoning. Frontiers in Microbiology, [online] 9(436), pp.1–18.
doi:https://doi.org/10.3389/fmicb.2018.00436.
• M. Tallent, S., W. Bennett , R. and M. Miller, W.B. (2023). BAM Chapter 13B - Staphylococcal Enterotoxins Detection Methods. FDA. [online] Available at: https://www.fda.gov/food/laboratory-methods-
food/bam-chapter-13b-staphylococcal-enterotoxins-detection-methods#:~:text=Staphylococcal%20enterotoxins%20(SEs)%20are%20pyrogenic [Accessed 3 Nov. 2023].
• Webb, B.H. (2023). Dairying. [online] Encyclopedia Britannica. Available at: https://www.britannica.com/topic/dairying.
• WebstaurantStore. (n.d.). WipesPlus 7" x 9" 100 Count No-Rinse Food Contact Surface Sanitizing Wipes. [online] Available at: https://www.webstaurantstore.com/wipesplus-7-x-9-food-contact-sanitizing-
wipes-canister/50033808.html [Accessed 5 Nov. 2023].
• Wu, S., Duan, N., Gu, H., Hao, L., Ye, H., Gong, W. and Wang, Z. (2016). A Review of the Methods for Detection of Staphylococcus aureus Enterotoxins. Toxins, [online] 8(7), p.176.
doi:https://doi.org/10.3390/toxins8070176.

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Staphylococcus aureas (1).pptx

  • 1. Staphylococcus aureus Foodborne pathogen: Staphylococcal food-borne disease Student name: Keerthana Kissanthomas Student Id No. :A00280414
  • 2. Introduction • Gram positive bacteria (cocci shaped that occur in clusters & member of Bacillota) • Commensal bacteria in human microbiota. • Opportunistic pathogen (common contributor to skin diseases, sinusitis, and food poisoning) Fig 1 – morphology of S. aureus under microscope after gram staining (Ganguly, 2018) Staphylococcal food-borne disease (SFP) results from the contamination of food by S. aureus’s enterotoxins (SE). (Kadariya, Smith and Thapaliya, 2014) Food intoxication (Staphyloenterotoxicosis) • S. aureus bacteria is killed by cooking, but its toxins are resistant to heat (withstand heat at 121°C for 10 min) and stomach enzymes (proteases). (M. Tallent, W. Bennett and M. Miller, 2022)
  • 3. Symptoms and Incubation period It is generally understood and accepted that S. aureus as a food intoxicant presents a risk only when the growth to high numbers occurs - >/= 100,000 CFU/g (>/= 1mg). (Rajkovic, 2016) Primary symptoms: Vomiting, nausea, diarrhoea and abdominal pain. Secondary symptoms : dehydration, headache, myalgia and fatigue Incubation period: Symptoms generally occurs between 30 minutes to 8 hours (average of 3 hours) after ingestion. Symptoms lasts 1-3 days. (CDC, 2018) Images generated by (Bio render 2023
  • 4. Food sources and transmissions • Prefers to deposit their enterotoxins on poultry, meats, diary, bakery products and salads. In raw foods, S. aureus does not compete effectively with indigenous bacteria. Source of contamination? Air, dust, and food contact surfaces can all act as carriers for S. aureus to enter foods. (Image by (WebstaurantStore, 2023) . Source: Argudín, Mendoza and Rodicio, 2010 Major source of contamination is poor handling of cooked or processed foods and followed by storage suitable. (Image by blog.userve.com, 2022) S. aureus is also found in dairy cattle (cows), particularly those with subclinical mastitis. (Image by Webb, 2023)
  • 5. Some Outbreaks Table 1: some examples of outbreaks of Staphyloenterotoxicosis (Hennekinne, De Buyser and Dragacci, 2012) Year Place, Country Food source Cases References
  • 6. Pathogenesis/virulence & Mode of action (i) Table 2: Main Staphylococcal enterotoxin (SE) of S. aureus involved in food (Pinchuk, Beswick and Reyes, 2010) •After the consumption, enterotoxins are absorbed in the abdomen and cause typical gastroenteritis. •Induces 5-hydroxytrytamine (5-HT) from mast cells which stimulates the vomiting centre of the brain •Enterotoxins also cause damage to the epithelial lining of the intestinal cells. The genes for enterotoxin production are present in pathogenicity islands in the chromosome (Arun K. Bhunia 2018) Source: Dura, 2021 Fig 2: S. aureus enterotoxins in stomach (Arun K. Bhunia 2018 citied in Dura, 2021) Fig 3: S. aureus enterotoxins in intestinal cells (Fisher E. L, et al. 2018)
  • 7. Pathogenesis/virulence & Mode of action (ii) • SEs are superantigens (SAgs) - stimulates many T-cells • Enterotoxins form complex with MHC class II molecules on surface of antigen- presenting cells • Proliferates T-cells, producing a large amounts of inflammatory cytokines (IL-2, IFNγ, IL-1, TNF-α). • A high abundance of inflammatory cytokines stimulates the neuron receptors in the intestinal tract and causes diarrhoea. Source: Dura, 2021 Fig 4: SEs as superantigen (Fisher E. L, et al. 2018) 1. 15-year-old (healthy) boy presented with septic shock after eating meat from a fast-food restaurant. 2. Developed food intoxication symptoms (diarrhoea) after 4 hours. 3. TSST-1 gene verified by real-time PCR analysis of the S.aureus strain. Source: Goudsmit et al., 2021 • Food poising from S. aureus cause toxic shock syndrome, a SAgs called toxic shock syndrome toxin 1 (TSST-1), is produced.
  • 8. Detection Source: Wu et al., 2016 Toxins not the bacterium itself
  • 9. Prevention • Prevent food from being held at an unsafe temperature (between 4.4°C and 60°C) for more than 2 hours. • Refrigerate as soon as possible (below 4 °C ). • Food handlers practicing good hygiene. • Avoid cross contamination (raw and cooked food). • If reheating food, ensure the temperature reaches at least 74 °C . Image by Evans, n.d shown in Pinterest n.d
  • 10. References • Argudín, M.Á., Mendoza, M.C. and Rodicio, M.R. (2010). Food Poisoning and Staphylococcus aureus Enterotoxins. Toxins, [online] 2(7), pp.1751–1773. doi:https://doi.org/10.3390/toxins2071751. • blog.userve.com. (2022). Food Handler How-To: Handle Poultry Properly. [online] Available at: https://blog.userve.com/us/food-handler-how-to-handle-poultry-properly [Accessed 5 Nov. 2023]. • Centers for Disease Control and Prevention (2018). Staphylococcal (staph) food poisoning. [online] Centers for Disease Control and Prevention. Available at: https://www.cdc.gov/foodsafety/diseases/staphylococcal.html. • Ganguly, S. (2018). Photograph showing S. aureus grape cluster like morphology on Gram stain. 1000X. [Image] ResearchGate. Available at: https://www.researchgate.net/publication/322243680_Molecular_Characterization_of_Staphylococcus_aureus_of_Camel_Camelus_dromedarius_Skin_Origin/figures?lo=1 [Accessed 30 Oct. 2023] • Kadariya, J., Smith, T.C. and Thapaliya, D. (2014). Staphylococcus aureus and Staphylococcal food-borne disease: An ongoing challenge in public health. BioMed Research International, [online] 2014, pp.1– 9. doi:https://doi.org/10.1155/2014/827965 • Rajkovic, A. (2016). Staphylococcus: Food Poisoning. [online] , Ghent University, Ghent, Belgium & Belgrade University, Zemun-Belgrade, Serbia: Elsevier Ltd, pp.133–139. Available at: https://elearning.unite.it/pluginfile.php/187875/mod_resource/content/0/rajkovic%2C2016-Staphylococcus-food%20poisoning.pdf [Accessed 2 Nov. 2023]. • Hennekinne, J.-A., De Buyser, M.-L. and Dragacci, S. (2012). Staphylococcus aureusand its food poisoning toxins: characterization and outbreak investigation. FEMS Microbiology Reviews, 36(4), pp.815– 836. doi:https://doi.org/10.1111/j.1574-6976.2011.00311.x. • Pinchuk, I.V., Beswick, E.J. and Reyes, V.E. (2010). Staphylococcal Enterotoxins. Toxins, [online] 2(8), pp.2177–2197. doi:https://doi.org/10.3390/toxins2082177. • Dura, S. (2021). Staphylococcal food poisoning (SFP)- Staphylococcus aureus enterotoxins. [online] Microbe Notes. Available at: https://microbenotes.com/staphylococcal-food-poisoning/ [Accessed 2 Nov. 2023]. • Fisher, E.L., Otto, M. and Cheung, G.Y.C. (2018). Basis of Virulence in Enterotoxin-Mediated Staphylococcal Food Poisoning. Frontiers in Microbiology, [online] 9(436), pp.1–18. doi:https://doi.org/10.3389/fmicb.2018.00436. • M. Tallent, S., W. Bennett , R. and M. Miller, W.B. (2023). BAM Chapter 13B - Staphylococcal Enterotoxins Detection Methods. FDA. [online] Available at: https://www.fda.gov/food/laboratory-methods- food/bam-chapter-13b-staphylococcal-enterotoxins-detection-methods#:~:text=Staphylococcal%20enterotoxins%20(SEs)%20are%20pyrogenic [Accessed 3 Nov. 2023]. • Webb, B.H. (2023). Dairying. [online] Encyclopedia Britannica. Available at: https://www.britannica.com/topic/dairying. • WebstaurantStore. (n.d.). WipesPlus 7" x 9" 100 Count No-Rinse Food Contact Surface Sanitizing Wipes. [online] Available at: https://www.webstaurantstore.com/wipesplus-7-x-9-food-contact-sanitizing- wipes-canister/50033808.html [Accessed 5 Nov. 2023]. • Wu, S., Duan, N., Gu, H., Hao, L., Ye, H., Gong, W. and Wang, Z. (2016). A Review of the Methods for Detection of Staphylococcus aureus Enterotoxins. Toxins, [online] 8(7), p.176. doi:https://doi.org/10.3390/toxins8070176.

Editor's Notes

  1. 2 secounds
  2. 30 secounds Staphylococcus aureus, a natural inhabitant of the human and animal body and reside on either surface of the body or at mucosa without harming human health. The best way to avoid food poisoning by Staph is to prevent food from being held at an unsafe temperature (between 4.4°C and 60°C) for more than 2 hours.
  3. 28 Salted food products, such as ham, have also been implicated, according to the capacity of S. aureus to grow at relatively low water activity (water activity = 0.86)
  4. 1m 20 secs Bakery products, particularly cream-filled pastries and cakes, and sandwich fillings. Food handlers carrying enterotoxin-producing S. aureus in their noses or on their hands are regarded as the main source of food contamination, via manual contact or through respiratory secretions
  5. Concntrare d in diary
  6. 25 different prophage encoded enterotoxins, SEA – SEE are the major reported for food poisoning Fig 2: the enterotoxin induces the release of 5-HT (5-hydroxytryptamine) from mast cells, which stimulates vagal nerves in the stomach lining and induces vomiting. Fig 3: The enterotoxins transit through mucus-expelling goblet cells and epithelial cells in the intestinal epithelium to reach the lamina propria. Here, the enterotoxins can interact with mast cells to induce the release of 5-hydroxytryptamine (5-HT/serotonin precursor), which interacts with the vagus nerve to cause an emetic response. Additional cellular targets that may have possible roles in the induction of enterotoxigenic disease include different types of T cells.
  7. Superantigens (SAgs) are a family of highly potent mitogens that share the ability to trigger excessive stimulation of human and other mammalian T lymphocytes. This leads to a massive release of T cell mediators and pro-inflammatory cytokines contributing to diseases such as toxic shock syndrome. Along with SEs, S. aureus also produces another super antigenic toxin (TSST-1).
  8. 3 types of Immunoassays.. 1)Optical detection techniques: Optical-based detection methods are widely applied in biomolecular analysis due to their ability to utilize various optical properties for detection, including light absorption, fluorescence/luminescence, chemiluminescence, Raman scattering, and refractive index changes. Optical immunoassays provide advantages such as speed, high sensitivity, real-time monitoring, and suitability for low sample concentrations and simplified sample preparation. These assays have found extensive use in the detection of SEs . 2) Electrochemical detection techniques: Electrochemical immunoassays offer a simple, sensitive, and cost-effective method for quantifying specific entities (SEs) based on changes in electric signals, including current, potential, impedance, and conductance. Researchers have developed various electrochemical detection methods for SEs, such as SEC1 and SEB, with low limits of detection. Signal amplification strategies have been employed to enhance sensitivity, including using polysilicon nanowires, horseradish peroxidase-nanosilica-doped multiwalled carbon nanotubes (HRPSiCNTs), and magnetosomes in combination with polydimethylimidazolium hexafluorophosphate ([D(n-C4)Im][PF6]) and polyaniline nano-gold composite (PANI/Au). These methods enable the detection of SEs at low concentrations, making them suitable for various applications, including the analysis of milk samples. 3) Mass detection techniques: Mass-based immunoassays rely on detecting small changes in mass to quantify specific entities (SEs). Piezoelectric (PZ) devices, such as quartz crystal microbalances, are used in these assays. When the analyte binds to the adsorbent on the crystal's surface, it increases the mass, causing a decrease in resonance frequency. Researchers have developed piezoelectric immunosensors for SEB and SEC2 detection, with sensitivities down to 0.1 µg/mL. Additionally, magnetoelastic sensors, similar to piezoelectric sensors, detect changes in resonance frequency due to mass loading when an analyte binds to a receptor immobilized on their surface. A magnetoelastic immunosensor for SEB detection achieved a detection limit of 0.5 ng/mL without the need for additional labels. These mass-based sensors offer high sensitivity and quick response times while minimizing costs. Drawback of immunoassays:  immunobioassays are a reliable and technology for SE detection, which exhibit high sensitivity, wide linear range and feasibility, they require high quality antibodies. The preparation of the antibodies via animal immunization experiments is tedious and time-consuming. Besides, the obtained antibodies may be unstable, and susceptible to modification issues