BIO 425 Industrial Microbiology

Lecture 9 Other pathogenic microorganisms Novel techniques to eliminate m/os HACCP

Content outline
Shigella spp. Listeria spp. Yersinia spp. Clostridum spp. Brucella spp. Mycotoxins Techniques to eliminate microorganisms HACCP concept

Shigella spp.
Differs from the E.coli by the lack of gas production, important genera; Shigella dysenteriae causes diarrhea, mild or severe, be watery or bloody. Symptoms are: fever and nausea, vomiting, abdominal cramps.

Listeria monocytogenes
Gr (+), non-spore former, less sensitive to heat can grow at T, as low as 3o C Present in both raw and processed food (mainly milk & cheese, seafood meat products, raw vegetables)

Listeria monocytogenes
Responsible for opportunistic infections, mainly in pregnant women, newborns and elderly.

Main symptoms of Listeriosis:

meningitis, encephalitis or septicaemia can lead to abortion, or premature birth when pregnant women are infected in the second and third trimesters.

Yersinia enterocolitica
Three pathogenic species of genus Yersinia is repeated. Cause gastroenteritis, rod shaped.

Yersinia pestis
the causative agent of the plague. Genetically very similar to Y. pseudotuberculosis. but infects humans by routes other than food.

Y. enterocolitica
detected in environmental and food sources such as ponds, lakes, meat, ice-cream and milk. Most isolates are reported to be not pathogenic.

Staphylococcus aureus
Gr (+), cocci; pathogenicity & food poisoning is through enterotoxins. (different types (7) varied by antigens, resistant to cooking and proteolytic enzymes), highly heatstable toxins. Exist in air, dust, sewage, water, milk & food, present in nasal passages and throat.

Foodstuff frequently invaded by S. aureus

meat & meat products, poultry & egg products, salads such as egg, tuna, chicken, potato and macaroni, cream-filled pastries, cream pies, sandwich fillings, milk & dairy products.

S. aureus poisoning is observed in:

foods that require considerable handling during preparation. and that are kept at slightly elevated temperatures after preparation: frequently involved in staphylococcal food poisoning.

Onset of symptoms include for S. aureus

1. Nausea, 2. Vomiting, 3. Abdominal cramps.

Clostridium perfringens
Anaerobic, G (+), Spore-former rod, Spores persists in soil, sediments. Areas subject to human or animal faecal position. Meat & meat products & gravy cheese are the foods most frequently implicated.

Clostridium perfringens
Five types (A-E) of Clostridium perfringens according to exotoxins. Common symptoms include abdominal pain, Nausea, acute diarrhea. They start after 8-12 hours of consumption of the toxin producing Clostridium perfringens.

Clostridium perfringens
Small no. of the organisms are often present as spores after cooking. The spores generate; and the Clostridia multiply to food poisoning levels; during the cooking period and storage.

Clostridium perfringens
The cooking process drives off oxygen, hence creating an anaerobic environment favorable to Clostridial growth. After ingestion: the enterotoxin is produced in the intestine, associated with sporulation, possibly induced by the acidic environment of the stomach.

Clinically important Clostridium spp.

C. botulinum C. perfringens C.difficile C.tetani C.botulinum and C.perfringens--> food poisoning

Botulism
through neurotoxins. They block the release of the neurotransmitter acetylcholine (AcH) results in muscle weakness & subsequent paralysis.

Botulism
Associated with canned food, (esp. Home canned) low acid foods, vegetables, meat and fish products. Also associated with honey; and hence honey should not be given to children under one year old: Infant botulism is milder than the adult version.

Bacillus cereus
Sporeformer, G(+), facultatively aerobic; survive during cooking process. They grow well in cooked food because of the lack of a competing microflora.

Brucella
Brucella melitensis (goats) Brucella abortus (cattle) Brucella suis (swine) strictly aerobic and Gr (-) coccobacilli-> cause Brucellosis, enters via skin, respiratory tract or the digestive tract.

Brucella
The intracellular organism can enter the blood and the lymphatics; where it multiplies inside phagocytes and eventually cause bacto-aemia (bacterial defection of blood); Includes high fever, chills and sweating; Pathogenicity: related to the production of lipopolysaccharides like poly N-formyl peroxamine O chain.

Mycotoxins
Produced by fungal genera; Aspergillus, Fusarium Penicillium.

Aflatoxins
Produced by Aspergillus spp. (B1,B2, G1, G2) by the blue or green fluorescence given under an UV lamp.), B1 is most toxic. Symptoms: acute necrosis, cirrhosis & carcinoma of liver, Potent carcinogen in birds, rodents, fish & human. Liver: the primary target of organism.

Methods of preservation and shelf life extension of food:

Pasteurization: applied to milk & some of fermented foods or juices. 630 C for 30 min. In milk HTST; high temp. short time, by 720 C for 15 sec. Designated to kill all pathogenic bacteria; e.g. Mycobacterium tuberculosis, Salmonella spp., Brucella spp.

More on pasteurization
Thermoduric organisms survive pasteurization; including; Streptococcus thermophilus Enterococcus faecalis Bacillus cereus -> its growth cause pasteurized milk spoilage known as bitty cream.

Sterilization
Milk sterilization (130o C at least 1 second.): a well established method for prolonged milk storage (UHT). (140o C for milisecond). The heat treatment: severe enough to kill all m/o present.

Sterilization
By heating: Both spoilage and foodborne pathogens: to an acceptable level. There is a statistical chance of an organism surviving the process, but this is acceptable: in the normal sense of safe food production.

High hydrostatic pressure (HHP)

High hydrostatic pressures of 300 to 500 MPa are applied: inactivate vegetative bacteria, whereas higher pressures (<1000 MPa) are required to kill bacterial spores. Novel method for food preservation.

Food irradiation:
Exposure of food, either packaged or in bulk to controlled amounts of ionization radiation (usually gamma rays from 60Co); for a specific period of time to achieve certain desirable objectives.

Food irradiation:
Presents the growth of m/o by damaging the DNA. It can also delay ripening and maturation by causing biochemical reactions in the plant tissues. limited to radiation from high energy rays, X-rays and accelerated electrons e.g. Microwave technology

Reduced oxygen packaging (ROP):

Its results in the food being surrounded by an atmosphere are observed: which contains little or no oxygen.

Controlled atmosphere packaging (CAP):

an active system in which a desired atmosphere is maintained for the duration: of the shelf life by the use of agents to bind; or scavenge oxygen or a gas generating kit.

Modified atmosphere packaging(MAP):

uses a gas flushing and sealing process; or reduction of oxygen through respiration of vegetables or microbial action. defined as packaging of a product in an atmosphere; which has had a one-time modification of gaseous composition; so that it is different from that air.

Modified atmosphere packaging (MAP)

defined as the enclosing of food products in an atmosphere inside gas-barrier materials, in which the gaseous environment has been modified to slow down; respiration rate, microbial growth; and reduce enzymatic degradation, with shelf life extension.

Modified atmosphere packaging(MAP)

Solid CO2 (dry ice) controls m/o growth by acting as: a refrigerant during transport and storage.

Vacuum packaging:
If the amount of air is reduced from a package; and hermetically seals it; so that a near-perfect vacuum remains inside.

Ultrasound:
Waves inactivate m/o by introducing alternating compression; and expansion cycles in a liquid medium. Small bubbles grow during the expansion of ultrasound. Temperature & pressure reached inside the bubbles is extremely high.

HACCP:
Hazard Analysis Critical Control Point

HACCP Implementation
late 1960s; the National Aeronautics and Space Administration (NASA) & U.S Army laboratories jointly developed the HACCP concept. then introduced to food industry. a simple but very specific way to find hazards in a food product. designed to prevent, rather than detect food hazards.

HACCP
looks at the flow of food through the process: provides a way to determine which points are critical for the control of foodborne disease hazards; and monitors each operation frequently.

HACCP combines two criteria:

1. A hazard is something that could harm the consumer. 2. A critical control point(CCP) is an operation or step where something can be done to prevent or control a hazard.

HACCP concept has two parts; (HACCP principles)

1. Hazard analysis depends on an understanding of which chemical and physical contaminants and m/o can be present and the factors that affect their growth and survival.

HACCP concept has two parts; (HACCP principles)

2. Critical control points are the points: where controls are needed to make sure that microbes do not reach the food, survive or grow to dangerous levels. Not all steps in a food production process are critical; and it is important to separate critical and non-critical points.

HACCP concept has two parts; (HACCP principles)

A flow diagram developed during hazard analysis can help to find CCP. Monitoring may take the form of observation, physical measurements (e.g. temperature, pH) or microbial analysis.

Factors most often affect food safety & acceptability

1.Contaminated raw food or ingredients. 2.Poor temperature control during processing and storage (time-temperature abuse). 3.Improper cooling(failure to control to refrigerated temperature within 2 hrs.). 4.Improper handling after processing.

Factors most often affect food safety & acceptability:

5.Cross-contamination. 6.Poor cleaning of the equipment. 7.Failure to separate raw and cooked products. 8.Poor employee hygiene and sanitation.

For HACCP to function well in the food industry:

1. Food processors and regulators must learn about HACCP. 2. Plant personnel must apply their technical skills to the program. 3. Sanitarians should drop HACCP controls points that are not hazardous from the program.

For HACCP to function well in the food industry

Development of HACCP in a food plant through good manufacturing practices (GMPs) & sanitary standard operating procedures(SSOPs) in accordance with Total Quality Management (TQM) & implementation of ISO 9000, 9001, 9002, 14000 and 22000.

Success with HACCP:

Employees in a food plant need special training to use HACCP. If all employees understand the important of keeping CCPs within the critical limits: cost effective control can replace costly crisis management; and outbreaks of foodborne illnesses can be reduced.

An effective HACCP program requires four steps;

1. Management education: Awareness & introduction of all staff via quality assurance. 2. Operational steps: Team must need to change plant design and operating system to take more hygienic measures.

An effective HACCP program requires four steps

3. Employee maturation: Good working conditions for maturating staff to carry out HACCP. 4. Employee involvement: Formation of quality circles among all levels of employees from boss to nonqualified any workers.

LITERATURE CITED
Forsythe, S.J. The Microbiology of Safe Food, Blackwell Science, Cambridge, 2000. kmen, Z. A. , Food Microbiology Laboratory Book, Food Engineering Department, Middle East Technical University, Ankara, 1995. Pelczar, M. J. ; Chan, E. C. S. ,Krieg, N. R. Microbiology, Fifth edition, Mc Graw Hill Inc. Singapore, 1986. Wood, B. J. B., Microbiology of Fermented Foods, Vol.1 and 2, Blackie Academic and Professional, London, Second edition, 1998.