Friday, May 23, 2008

Staphylococcus Aureus

Staphylococcus aureus


S.aureus is a gram positive coccus, which appears as grape-like clusters when viewed through a microscope and has large, round, golden-yellow colonies, S.aureus is a commensal of warm blooded animals (Commensal bacteria are normal bacteria which make up part of the mucosal flora (i.e. the mouth, the nose, the lungs, and the gastrointestinal and urogenital tracts) of animals but do not cause disease.)

S.aureus can produce a range of substances including enterotoxins (toxin produced by bacteria that is specific for intestinal cells and causes the vomiting and diarrhoea associated with food poisoning) and haemolysins (the destruction of red blood cells with release of haemoglobin

Food poisoning occurs when the growth of S.aureus in a food reaches about 1 million (106 cfu/g) and toxins have formed in the food.

Contamination of food with S.aureus/S.aureus toxin is usually caused by food handlers. Can be a result of poor handling techniques or such as a food handler with open sores, spots or lesions.

Symptoms occur within 1 to 8 hours after ingestion of the contaminated food.

Symptoms include nausea, vomiting and abdominal cramps

Usually self limiting and clears up without any specific treatment

Staphylococcal food poisoning is not an infection, but an intoxication that results from ingestion of staphylococcal enterotoxins in food. The enterotoxins are produced when food contaminated with S. aureus is improperly stored under conditions that allow the bacteria to grow. Although contamination can originate from animals or the environment, food handlers with poor hygiene are the usual source. Effective methods for preventing staphylococcal food poisoning are aimed at eliminating contamination through common hygiene practices, such as wearing gloves, and proper food storage to minimize toxin production.

S.aureus is resistant to drying which makes it difficult to control (eg hand washing and drying not always effective)

S.aureus can colonise on equipment if it is left wet (and biofilms may also occur will protect the organism even more)

S.aureus is quickly destroyed by adequate cooking but if enterotoxins have already formed, these can be resistant to heat.

S.aureus are able to grow and produce enterotoxin in low water activity foods
environment is another source of contamination

S.aureus are resistant to freezing and thawing. The enterotoxin is not affected by freezing (problems with ice cream).

Heat resistance is increased in high fat and dry foods, so increased cooking requirements must be put into place when preparing theses type of products.

They are tolerant to salt (can cause a problem with foods such as cooked hams, etc)

They can grow in acidic conditions (pH <4.3)

Can grow aerobic and anaerobically

Salmonella

Salmonella






Salmonella is a Gram-negative facultative rod-shaped, the family Enterobacteriaceae, Salmonellae live in the intestinal tracts of warm and cold blooded animals. Some species are ubiquitous. Other species are specifically adapted to a particular host. In humans, Salmonella are the cause of two diseases called salmonellosis: enteric fever (typhoid), resulting from bacterial invasion of the bloodstream, and acute gastroenteritis, resulting from a foodborne infection/intoxication.

Habitats

The principal habitat of the salmonellae is the intestinal tract of humans and animals. Salmonella serovars can be found predominantly in one particular host, can be ubiquitous, or can have an unknown habitat. Typhi and Paratyphi A are strictly human serovars that may cause grave diseases often associated with invasion of the bloodstream. Salmonellosis in these cases is transmitted through fecal contamination of water or food. In human adults, ubiquitous Salmonella organisms are mostly responsible for foodborne toxic infections.

Salmonella in the Natural Environment Salmonellae are disseminated in the natural environment (water, soil, sometimes plants used as food) through human or animal excretion. Humans and animals (either wild or domesticated) can excrete Salmonella either when clinically diseased or after having had salmonellosis, if they remain carriers. Salmonella organisms do not seem to multiply significantly in the natural environment (out of digestive tracts), but they can survive several weeks in water and several years in soil if conditions of temperature, humidity, and pH are favorable


Salmonellae survive sewage treatments if suitable germicides are not used in sewage processing. In a typical cycle of typhoid, sewage from a community is directed to a sewage plant. Effluent from the sewage plant passes into a coastal river where edible shellfish (mussels, oysters) live. Shellfish concentrate bacteria as they filter several liters of water per hour. Ingestion by humans of these seafoods (uncooked or superficially cooked) may cause typhoid or other salmonellosis. Salmonellae do not colonize or multiply in contaminated shellfish.
Typhoid is strictly a human disease.The incidence of human disease decreases when the level of development of a country increases (i.e., controlled water sewage systems, pasteurization of milk and dairy products). Where these hygienic conditions are missing, the probability of fecal contamination of water and food remains high and so is the incidence of typhoid.
Foodborne Salmonella toxic infections are caused by ubiquitous Salmonella serovars (e.g., Typhimurium). About 12-24 hours following ingestion of contaminated food (containing a sufficient number of Salmonella), symptoms appear (diarrhea, vomiting, fever) and last 2-5 days. Spontaneous cure usually occurs.

Salmonella may be associated with all kinds of food. Contamination of meat (cattle, pigs, goats, chicken, etc.) may originate from animal salmonellosis, but most often it results from contamination of muscles with the intestinal contents during evisceration of animals, washing, and transportation of carcasses. Surface contamination of meat is usually of little consequence, as proper cooking will sterilize it (although handling of contaminated meat may result in contamination of hands, tables, kitchenware, towels, other foods, etc.). However, when contaminated meat is ground, multiplication of Salmonella may occur within the ground meat and if cooking is superficial, ingestion of this highly contaminated food may produce a Salmonellainfection. Infection may follow ingestion of any food that supports multiplication of Salmonella such as eggs, cream, mayonnaise, creamed foods, etc.), as a large number of ingested salmonellae are needed to give symptoms. Prevention of Salmonella toxic infection relies on avoiding contamination (improvement of hygiene), preventing multiplication of Salmonella in food (constant storage of food at 4°C), and use of pasteurized and sterilized milk and milk products. Vegetables and fruits may carry Salmonella when contaminated with fertilizers of fecal origin, or when washed with polluted water.

The incidence of foodborne Salmonella infection/toxication remains reletavely high in developed countries because of commercially prepared food or ingredients for food. Any contamination of commercially prepared food will result in a large-scale infection. In underdeveloped countries, foodborne Salmonella intoxications are less spectacular because of the smaller number of individuals simultaneously infected, but also because the bacteriological diagnosis of Salmonella toxic infection may not be available. However, the incidence of Salmonella carriage in underdeveloped countries is known to be high.

Salmonella epidemics may occur among infants in pediatric wards. The frequency and gravity of these epidemics are affected by hygienic conditions, malnutrition, and the excessive use of antibiotics that select for multiresistant strains.
Salmonella Enteritidis Infection Egg-associated salmonellosis is an important public health problem in the United States and several European countries. Salmonella Enteritidis, can be inside perfectly normal-appearing eggs, and if the eggs are eaten raw or undercooked, the bacterium can cause illness.

Unlike eggborne salmonellosis of past decades, the current epidemic is due to intact and disinfected grade A eggs. Salmonella Enteritidis silently infects the ovaries of healthy appearing hens and contaminates the eggs before the shells are formed. Most types of Salmonella live in the intestinal tracts of animals and birds and are transmitted to humans by contaminated foods of animal origin. Stringent procedures for cleaning and inspecting eggs were implemented in the 1970s and have made salmonellosis caused by external fecal contamination of egg shells extremely rare. However, unlike eggborne salmonellosis of past decades, the current epidemic is due to intact and disinfected grade A eggs. The reason for this is that Salmonella Enteritidis silently infects the ovaries of hens and contaminates the eggs before the shells are formed.
A person infected with the Salmonella Enteritidis usually has fever, abdominal cramps, and diarrhea beginning 12 to 72 hours after consuming a contaminated food or beverage. The illness usually lasts 4 to 7 days, and most persons recover without antibiotic treatment. However, the diarrhea can be severe, and the person may be ill enough to require hospitalization. The elderly, infants, and those with impaired immune systems (including HIV) may have a more severe illness. In these patients, the infection may spread from the intestines to the bloodstream, and then to other body sites and can cause death unless the person is treated promptly with antibiotics.

Exotoxins
Salmonella strains may produce a thermolabile enterotoxin that bears a limited relatedness to cholera toxin both structurally and antigenically. This enterotoxin causes water secretion in rat ileal loop and is recognized by antibodies against both cholera toxin and the thermolabile enterotoxin (LT) of enterotoxinogenic E. coli, but it does not bind in vitro to ganglioside GM1 (the receptor for E. coli LT and cholera ctx). Additionally, a cytotoxin that inhibits protein synthesis and is immunologically distinct from Shiga toxin has been demonstrated. Both of these toxins are presumed to play a role in the diarrheal symptoms of salmonellosis.

Tuesday, May 20, 2008

Food Microbiology

Most of the micro-organisms associated with food microbiology fall into one of the following groups -
Viruses
Bacteria
Parasites
Fungi

Viruses – much smaller than bacteria. Cannot be seen by normal light microscopy. We would need to use an Electron microscope.

A virus is made up of a core of nucleic acid surrounded by a protein coat.

Cannot grow in food but lay dormant in food and water. When ingested they can become active.

An example of a virus is Hepatitis A (associated with shellfish). Shellfish are filter feeders and tend to live near sewage outfall. This concentrates the viruses which may be in the shellfish. This may present a problem for example for people eating raw oysters.

Norovirus
Lace-like appearance of individual virus particles. The virus particles of Norwalk and other
SRSVs (Small Round Structured Viruses)

Rotavirus
Wheel-like appearance. The observance of such particles gave the virus its name
('rota' being the Latin word meaning wheel).

Rotavirus more commonly causes vomiting and illness in children. It is thought that most people build a resistance to it.



Bacteria

Bacteria are minute, they cannot be seen without a microscope.

Only a small proportion are pathogenic most are Commensal flora. There are thousands of different types. Many perform useful functions.

One of the most distinguishing features of bacteria is their reaction to Grams stain. In 1884, Hans Christian Gram, a Danish doctor working in Berlin, accidentally stumbled on a method which still forms the basis for the identification of bacteria. While examining lung tissue from patients who had died of pneumonia, he discovered that certain stains were preferentially taken up and retained by bacterial cells. Over the course of the next few years, Gram developed a staining procedure which divided almost all bacteria into two large groups - the Gram stain

Gram stains fall into two groups – positive and negative
This is just a way of categorising bacteria.

Round shapes are cocci
Rod shaped are bacilli

Some bacteria are motile in fluids and can swim or dart about by means of flagella
Polar flagella at the end of the organism – these bacteria dart about ( a bit like tadpoles)
Bipolar – flagella at both ends, so they tumble about.
All round flagella – peritrichous – so they move about like a corkscrew or rifle bullet.

All bacteria are classified to genus and species eg Staphyloccocus aureus – Staphyloccocus is the genus, aureus is the species.


Other important cell components (not in all bacteria) include capsules and spores.

Capsules form a protective covering. Spores are formed when conditions are not favourable (eg lack of moisture). Spores are formed within the bacterial cells (endospores). When cell disintegrates, the spores remain intact. Can withstand heat, light, chemicals, lack of moisture etc. germinate when conditions are favourable.

Bacteria divide by simple division (into 2). Divide by binary fission and each generation is doubled in number (2,4,8,16,32 etc). Under suitable conditions may occur every 15 to 20 mins (therefore > million in 4 hours)?


Growth occurs in 3 stages

Lag – can grow but slowly – a period of adaptation to the environment
Logarithmic/exponential – constant specific growth rate (binary fission). Dependent on conditions, temp, time etc

Stationary – one or more of the nutrients becomes exhausted or toxic metabolic products accumulate and inhibit growth. Growth ceases. Number of viable bacteria remains constant

Decline/death – death rate exceeds the rate of multiplication. Majority of cells die. Some may form spores.


Temperature - important factor as it influences the rates of all chemical reactions linked to the process of growth. The temperature at which a culture medium is incubated determines the rate of growth of any bacteria associated with it. For any organism the temperature at which growth is most rapid is known as the optimum temperature.
The minimum temperature is the lowest temperature at which growth occurs (usually substantially below the optimum temperature)
The maximum temperature is the highest temperature at which the organism will grow. Usually only a few degrees above the optimum.

Bacteria can be distinguished on the basis of their temperature relationships
Psychrophiles – grow at very low temperatures. Optimum 5-20C and ability to grow at 0C (and as low as –22C). Rare group but include some spoilage bacteria (eg some species of pseudomonas)

Psychrotrophs - Yersinia enterocolitica is a psychrotrophic organism (can grow at fridge temps)
Listeria another psychrotrophic organism

Theromphiles prefer 45 to 70C

Hyperthermophiles – above 75C

Mesophiles – most food borne pathogens are mesophiles and grow best at 30 to 45C


Moisture – at least 80% bacterial cell consists of water. Essential for growth and survival
Bacteria vary on the dependency on water.

Atmosphere – Bacteria differ in their need for oxygen for growth, or in its exclusion. Bacteria that are dependent on free oxygen present in air are called obligate or strict aerobes. (eg Pseudomonas spp)
Bacteria that will only grow in the absence of oxygen are called obligate anaerobes. Even traces of Oxygen are toxic to them (eg Clostridia spp)
Majority of bacteria fall between these two extremes and are called facultative anaerobes (eg E coli, S aureus). Able to grow anaerobically but prefer aerobic conditions.

AEROBIC – grows in air
MICROAEROPHILIC – grows in reduced oxygen
FACULTATIVE – grows with or without oxygen
ANAEROBES – cannot grow if oxygen is present

PH – or hydrogen ion concentration. Significant effect on the growth of bacteria. All bacteria have an optimum pH. Most bacteria favour a neutral pH (6.8 to 7.5). Some prefer alkaline conditions (eg vibrio like 8.5 to 9.0, yeasts and moulds like alkaline conditions)

Time – most bacteria can form cfu within 18hours on agar under optimum conditions. But some need 48hrs before producing typical morphology. (in fact Legionella can take up to 10 days to grow – useful as contaminants can grow in 24 hours). Moulds can take up to 5 days

Micro-organisms are everywhere

Not all of them are pathogenic. Some of them are very useful. bacterial growth and survival depends on several factors. These include nutrients, pH, time, temperature.
Bacteria are able to survive harsh conditions (eg drying – think of spices). This can make them very difficult to control.


Biofilms: Biofilms are another way in which bacteria can protect themselves under harsh conditions. Biofilms can be very difficult to remove I have given you some example of biofilms on the slide

Biofilms are a problem on rough surfaces (think about chopping boards, serving areas)
Biofilms are of an organic nature (ie they are made up of bacteria) which may or may not be coated in substances such as food. They may lie dormant waiting for a trigger (eg moisture, warmth, food) to make them grow.


The micro-organism in a food does not have to grow to cause problems.
Some micro-organisms use the food as a vehicle of transmission

Some example of these are Viruses (the chef may have a viral infection – he is suffering from vomiting – he manages to get some of the virus onto the food he is preparing – his customers also become infected with the virus)
Another example are parsites such as Giardia which may be passed on via food from poor hygiene in the kitchen (eg an infected food handler does not wash his hands properly after going to the toilet)
Campylobacter does not grow in food but survives well. In the laboratory were I work we are finding that about 90% raw chicken is infected with campylobacter.


If the conditions are favourable (temperature, humidity, etc), and the organism is a pathogen
We may get food poisoning We can get food poisoning by an infective dose (ie we actually eat the bacteria in sufficient numbers to cause illness – this can be a very low number in some organisms such as E.coli O157) Or, we can become ill by ingesting the toxin which has been produced by the bacteria in the food (eg Staphylococcus aureus toxin which can cause severe vomiting within hours)

Friday, May 9, 2008

Food Safety Managment

Food Safety Management


Food controls can be traced back many centuries, with well documented rules governing the sale of bread and beer promulgated in the Middle Ages. These earliest statutory controls were, however, directed toward prevention of revenue evasion rather than protecting ‘trader’ or ‘public’, and legislation in its present form is very much a product of the nineteenth century. This period saw the rapid development of food legislation in an attempt to remedy the widespread practice of adulteration. Whilst not a new phenomenon the rapid urbanization of this time vastly increased the numbers of people reliant on manufacturers and retailers. In the absence of effective control of food quality the opportunities for adulteration were tremendous and the potential profits substantial.

Food was often adulterated with harmful ingredients, tea for example was extended with sawdust or blackened with lead, and coffee had chicory added. The damage to public health and the scandals caused by adulterated foods created the conditions that led to the first of the modern laws to control food quality. The identification and growing understanding of microbial pathogens however was not developed until the latter part of the 19th century following the work of Pasteur. This paved the way for effective legislative control of food safety, which began with milk pasteurization and water chlorination, and continues today in the form of extensive food safety regulations.

In the 1950’s when NASA scientists were developing the first manned space program they faced the problem of how to ensure the safety of food eaten by the astronauts. Originally they were concerned with the physical form of the food itself and whether the astronauts would choke on the crumbs in zero-gravity, or if the food materials might drift into the delicate machinery on the spacecraft

NASA was seeking an approach through which they could achieve 100% assurance of food safety and soon realized that traditional end-product testing was not going to accomplish this. Given the aim of ‘zero-defects’ to be achieved, any system of random testing would lead to very high losses of product, uncertainty and inadequacy in achieving safety due to the non-uniform nature of contamination in food products. For end-product testing to be useful it would require analysis of 100% of the product. An alternative system of safety assurance rather than safety control was needed.

HACCP

Hazard Analysis critical Control Point is widely recognized in the food industry as an effective approach to establishing production, Sanitation and manufacturing practices that produce safe foods.

HACCP systems establish process control by identifying points in production that are most critical to food system, and determine how these should be monitored and controlled. HACCP has often been expanded to include quality parameters.

The key to successful HACCP is proper preparation and planning, including gaining commitment at senior level and ensuring proper allocation of resources, including for HACCP specific training. Training will ensure the correct application and maintenance of the system. The key elements needed to set up an effective management system:


Transfer or ownership of HACCP plan to operatives, supervisor, and managers
Training of operatives, supervisors and managers to implement HACCP
Maintenance of HACCP plan

The development of HACCP system involves a number of stages a number of stages, the first of which is to set up a team of relevant people.


Pre-Requisites

The plan or layout must be designed to achieve the smooth flow of operations, and equipments should also be designed to facilitate cleaning, maintenance and inspection

Good Manufacturing Practices (GMP): This includes mechanisms in place to prevent cross-contamination through.

Supplier Quality Assurance: Having system in Place that allow confidence that supplier are producing and delivering safe foods consistently, based on criteria set by the purchasing company. Criteria may include product specifications of acceptable levels of contamination or that the supplier also operates a HACCP system and routine audits of suppliers.

Calibration: A system in place that ensure regular calibration of equipment, including monitoring equipment to ensure that such equipment is operating correctly

Good laboratory Practice: this includes the accreditation of systems used in the laboratory whether the laboratory is internal or external to the company. Other issues will include the training for staff, monitoring of staff performance, and building control into sample testing procedure.

Incident Management: The existence of system to be followed in the event of a serious incident. Such as a system would specify the actions to be taken by whom
What should happen to the affected production
Who should be informed and the procedures for investigation of the cause of the incident

Personnel and training: policies relating to the nature of the staff who should be employed to certain positions, the competence that they will need, and the training program available to develop, refresh and update such competencies. A policy may also include ways in which training needs are identified, including encouraging personnel to identify their own needs, mechanisms for establishing the effectiveness of training and maintenance of training records


Preventative Maintenance: A system in place that ensures regular preventative maintenance of premises and equipment. This may include contracts with pest control agencies to regularly audit premises for signs of infestations, as well as ensuring that such pests are not able to entre premises.