Microbiology and Engineering

2009.4.9

1. INTRODUCTION Because human food sources are of plant and animal origin, it is important to understand the biological principles of the microbial biota associated with plants and animals in their natural habitats and respective roles. Although it sometimes appears that microorganisms are trying to ruin our food sources by infecting and destroying plants and animals, including humans, this is by no means their primary role in nature. In our present view-of life on this planet, the primary function of microorganisms in nature is selfperpetuation. During this process, the heterotrophs and autotrophs carry out the following general reaction: All organic matter (carbohydrates, proteins, lipids, etc.) Energy + Inorganic compounds (nitrates, sulfates, etc.)

It is essentially nothing more than the operation of the nitrogen cycle and the cycle of other elements. The microbial spoilage of foods may be viewed simply as an attempt by the food biota to carry out what appears to be their primary role in nature. Microbiology fall into four categories; bacteria, yeasts, molds1, viruses.
BACTERIA MICRO BIOLOGY YEAST MOLD VIRUS

Bacteria is the most common food-borne pathogens. Bacteria growth rates, under optimum condition, are generally faster than those of the yeasts and molds. Below is a few list of important microorganisms the makes foodborne occurs.

A large group of fungi (like penicillium) that cause mold (as on bread or cheese). A common trigger for

allergies (http://www.biology-online.org/dictionary/Molds)

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2.

MICROORGANISMS SOURCE IN THE FOOD The microorganisms live in many medium such as air, water, soil, plant materials, and the skin J expected to persist in soils. The bacterial biota of seawater is essentially Gram-negative, and Gram-positive bacteria exist there essentially only as transients. Contaminated water has been implicated in Cyclospora contamination of fresh raspberries., Plants and Plant Products. It may be assumed that many or most soil and water organisms contaminate plants. However, only a relatively small number find the plant environment suitable to their overall well-being. Those that persist on plant products do so by virtue of a capacity to adhere to plant surfaces so that they are not easily washed away and because they are able to obtain their nutritional requirements. Notable among these are the lactic acid bacteria and some yeasts. Among others that are commonly associated with plants are bacterial plant pathogens in the genera Corynebacterium, Curtobacterium, Pectobacterium, Pseudomonas, and Xanthomonas; and fungal pathogens among several genera of molds. Food Utensils. When vegetables are harvested in containers and utensils, one would expect to find some or all of the surface organisms on the products to contaminate contact surfaces. As more and more vegetables are placed in the same containers, a normalization of the microbiota would be expected to occur. In a similar way, the cutting block in a meat market along with cutting knives and grinders are contaminated from initial samples, and this process leads to a buildup of organisms, thus ensuring a fairly constant level of contamination of meat-borne organisms. Gastrointestinal Tract. This biota becomes a water source when polluted water is used to wash raw food products. The intestinal biota consists of many organisms that do not persist as long in waters as do others, and notable among these are pathogens such as salmonellae. Any or all of the Enterobacteriaceae may be expected in fecal wastes, along with intestinal pathogens, including the five protozoal species already listed. Food Handlers. The microbiota on the hands and outer garments of handlers generally reflect the

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environment and habits of individuals, and the organisms in question may be those from soil, water, dust, and other environmental sources. Additional important sources are those that are common in nasal cavities, the mouth, and on the skin, and those from the gastrointestinal tract that may enter foods through poor personal hygiene practices. Animal Feeds. This is a source of salmonellae to poultry and other farm animals. In the case of some silage, it is a known source of Listeria monocytogenes to dairy and meat animals. The organisms in dry animal feed are spread throughout the animal environment and may be expected to occur on animal hides. Animal Hides. In the case of milk cows, the types of organisms found in raw milk can be a reflection of the biota of the udder when proper procedures are not followed in milking and of the general environment of such animals. From both the udder and the hide, organisms can contaminate the general environment, milk containers, and the hands of handlers. Air and Dust. Although most of the organisms listed in Table 22 may at times be found in air and dust in a food-processing operation, the ones that can persist include most of the Gram-positive organisms listed. Among fungi, a number of molds may be expected to occur in air and dust, along with some yeasts. In general, the types of organisms in air and dust would be those that are constantly reseeded to the environment. Air ducts are not unimportant sources see the table below; Molds are filamentous fungi that grow in the form of a tangled mass that spreads rapidly and may cover several inches of area in 2 to 3 days. The total of the mass or any large portion of it is referred to as mycelium. Mycelium is composed of branches or filaments referred to as hyphae. Those of greatest importance in foods multiply by ascospores, zygospores, or conidia. The ascospores of some genera are notable for their extreme degrees of heat resistance. One group forms pycnidia or acervuli (small, flask-shaped, fruiting bodies lined with conidiophores). Arthrospores result from the fragmentation of hyphae in some groups

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Table 1. eight important source of bacteria

Note: XX indicates a very importance source a Primary water b Primary soil c Nontuberculous

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Picture 1. Moldy bread2 Yeasts may be viewed as being unicellular fungi in contrast to the molds, which are multicellular; however, this is not a precise definition, as many of what are commonly regarded as yeasts actually produce mycelia to varying degrees. Yeasts can be differentiated from bacteria by their larger cell size and their oval, elongate, elliptical, or spherical cell shapes. Typical yeast cells range from 5 to 8 m in diameter, with some being even larger. Older yeast cultures tend to have smaller cells. Most of those of importance in foods divide by budding or fission. Yeasts can grow over wide ranges of acid pH and in up to 18% ethanol. Many grow in the presence of 5560% sucrose. Many colors are produced by yeasts, ranging from creamy, to pink, to red. The asco- and arthrospores of some are quite heat resistant. (Arthrospores are produced by some yeast-like fungi.)

http://en.wikipedia.org/wiki/File:Verschimmeltes_Brot_2008-12-07.JPG

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Picture 2. Dried yeasts3 3. MICROBIAL GROWTH The growth of microbial population can be generalize by using the curve below (see pic.3). an initial lag phase occurs as organism start to grow and adapt to new environmental condition. The lag phase is very important because the maximum extensions of shelf life and length of production run are directly related to the length of the lag phase. Once adaption has occurred, the culture enters into the maximum (logarithmic) growth rate, and control the microbial growth is not possible without major sanitation or other drastic measures. Numbers can double as fast as 20 to 30 min under optimum conditions. Toxin production and spore maturation, if possible, usually occur at the end at the end exponential phase as the culture enters the stationary phase. At this time, essential nutrients are depleted and/or inhibitory by-products are accumulated. Eventually the culture dies, the rate depending on the organism, the medium, and the other environmental characteristics.

http://en.wikipedia.org/wiki/File:Dry_yeast.jpg

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Number of cell logarithmic scale

stationary

h deat

lag

time

picture 3. Typical microbial growth curve4 The growth of microorganism has their own factors which are divided into two categories, first is intrinsic factor that are function of the food itself and second is the extrinsic factor that are the function of environmental in which the food is held.

4. INTRINSIC FACTOR The parameters of plant and animal tissues that are an inherent part of the tissues are referred to as intrinsic parameter. These parameters are as follows: 1. 2. 3. 4. 5. 6. Nutrient content Inhibitor pH Competing organism water activity Biological structures

Each of these substrate-limiting factors is discussed below, with emphasis placed on their effects on microorganisms in foods. 4.1. Nutrient

1994 ASHRAE Refrigeration Handbook, p.9.1

log

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Like other living organism, microorganisms require food to grow. Carbon and energy source are usually supplied in the form of sugars and starches. Nitrogen requirements are met by the presence of protein. Vitamins and minerals are also necessary. Lactic acid bacteria have rather exacting nutritional requirements, while may aerobic spore formers have tremendous enzymatic capabilities and are capable of growth on the wide variety of substrates. Cleanable system facilitate the removal of residual food material and deprive microorganisms of the nutrients required for growth, thus preventing a buildup of organisms in the environment 4.2. Inhibitors Inhibitor may be present in the food by naturally or addicted with preservatives. Preservatives are not substitute for hygienic practices and, with time, microorganisms may develop resistance. A cleanable system is still essential in preventing the development of resistance population.

4.3. Competing Microorganisms The presence of other microorganisms also affect the organisms in foods. Some organisms produce inhibiting compounds or has fast generation times, other are better able to use the available nutrients in a food matrix. 4.4. Water Activity One of the oldest methods of preserving foods is drying or desiccation; precisely how this method came to be used is not known. The preservation of foods by drying is a direct consequence of removal or binding of moisture, without which microorganisms do not grow. It is now generally accepted that the water requirements of microorganisms should be described in terms of the water activity (aw) in the environment. This parameter is defined by the ratio of the water vapor pressure of food substrate to the vapor pressure of pure water at the same temperature: aw = p/po, where p is the vapor pressure of the solution and po is the vapor pressure of the solvent (usually water). This concept is related to relative humidity (RH) in the following way: RH = 100 aw.13 Pure water has an aw of 1.00, a 22% NaCl solution (w/v) has an aw of 0.86, and a saturated solution of NaCl has an aw of 0.75 (Table 2).

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Table 2. relationship between water activity and concentration salt solution

The water activity (aw) of most fresh foods is above 0.99. The minimum values reported for the growth of some microorganisms in foods are presented in Table 3. In general, bacteria require higher values of aw for growth than fungi, with Gram-negative bacteria having higher requirements than Gram positives. Most spoilage bacteria do not grow below aw = 0.91, whereas spoilage molds can grow as low as 0.80

Table 3. Approximate Minimum aw Values for Growth of Microorganisms Important in Foods5

James M Jay,2005; Modern food microbiology 7th edition,

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4.5. PH It has been well established that most microorganisms grow best at pH values around 7.0 (6.67.5), whereas few grow below 4.0 . Bacteria tend to be more fastidious in their relationshipsto pH than molds and yeasts, with the pathogenic bacteria being the most fastidious. With respect to pH minima and maxima of microorganisms, those represented in picture 4, should not be taken to be precise boundaries, as the actual values are known to be dependent on other growth parameters. For example, the pH minima of certain lactobacilli have been shown to be dependent on the type of acid used, with citric, hydrochloric, phosphoric, and tartaric acids permitting growth at a lower pH value than acetic or lactic acids

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Picture 4. Approximate pH growth ranges for some food-borne organisms

5. EXTRINSIC FACTORS The extrinsic factor of that influence the growth of microorganisms are temperature, environmental relative humidity, and oxygen levels. Refrigeration and ventilation system play a major role in controlling these factors. 5.1. Temperature. Microorganisms are capable of growth over a wide range of temperature. Minimum growth temperatures for a variety of spoilage and pathogenic bacteria of significance in food are summarize in table 4. Table 4. minimum growth temperature for some bacteria in foods
Organism Possible significant Approximate min growth o temp, C.

Staphylococcus aureus

Food-born disease

10

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Salmonella spp. Clostridium botulinum proteolic nonproteolic Lactobacillus and leuconostoc Listeria monocytogenes Acinetobacter spp

Food-born disease Food-born disease

5.5

10 3 Spoilage of cooked sausage 3

Food-borne disease Spoilage of precooked food Spoilage of raw fish, meats, poultry, and dairy products

1 -1

Pseudomonads

-1

The lowest temperature at which a microorganism has been reported to grow is 34C; the highest is somewhere in excess of 100C. It is customary to place microorganisms into three groups based on their temperature requirements for growth. Those organisms that grow well at or below 7C and have their optimum between 20C and 30C are referred to as psychrotrophs. Those that grow well between 20C and 45C with optima between 30C and 40C are referred to as mesophiles, whereas those that grow well at and above 45C with optima between 55C and 65C are referred to as thermophiles. 5.2. Environmental relative humidity As previously discussed as an essential intrinsic growth factor, is also major extrinsic factor. Environmental water acts as a vector for transmission of microorganisms from one location to another through foot traffic or aerosols.

5.3. Oxigen

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AEROBIC

Microorganisms classified by oxygen requirement

ANAEROBIC

By controlling the oxygen level to prolong storage life by retarding growth of spoilage organisms in addition to influencing ripening processes. Vacuum packing of food also uses this extrinsic growth factor by inhibiting the growth of strict aerobes. 6. DESIGN FOR CONTROL OF MICROORGANISMS
Water control

Prevention of Contamination Prevention of Growth Destruction of Organisms

sanitation Prevention of Growth Humidity control

DESIGN

freezing

Picture 5. Food temperature chart6 High temperatures destroy most bacteria. The higher the temperature,

Taken from http://www.doh.state.fl.us/Environment/community/food/temp.htm

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the quicker and more effective the kill. Low temperatures prevent bacteria from reproducing. Bacteria thrive in a temperature range from 5 degrees C to 60 oC. This range includes room temperature and body temperature. In this range bacteria reproduce wildly and have the greatest potential for contamination spread and infection. Hot foods are generally cooked between 62.7 and 100 oC. It is important that center of grilled meat reaches at least 62.7 oC. Verify this with a meat thermometer. The minimum storage temperature for hot food is 160 degrees C. Cold foods should be kept at a range of -17.7 to 5 oC. The maximum storage temperature for cold food is 5 oC. Frozen foods should be kept frozen between -17.7 and - 34 oC. In the ASHRAE standard 52.1-1992, dust has to be suppressed to prevent the contamination of food, food is expose with air, the filtration of air is important by using 95% filters that dust would be sufficiently removed from the contamination. HEPA (high-efficiency particulate air) filters provide sterile air and are used for cleanrooms.

Picture 6. HEPA filter7

Taken from http://en.wikipedia.org/wiki/File:HEPA_Filter_diagram_en.svg

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