CHAPTER 16

MIXED CULTURES
 INTRODUCTION

• The dynamics of mixed cultures are

important considerations in some
commercial fermentations.
• The use of organisms with
recombinant DNA has added another
dimension to our consideration of
how cells within a population interact
with each other.

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16.2. MAJOR CLASSES OF INTERACTIONS
IN MIXED CULTURES

1. Competition
2. Neutralism
3. Mutualism
4. Commensalism
5. Amensalism
6. Prey–predator

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 COMPETITION

• Indirect interaction between two populations that has negative effects on

both.

• Two populations or microorganisms with similar nutrient requirements

usually compete for a number of common, required nutrients when grown
together.

• Exclusion Principle: the organisms with the fastest growth rate will displace
the others from the culture.

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 NEUTRALISM

• Is an interaction where neither population is affected by the presence of the

other.

• There is no change in the growth rate of either organism due to the presence
of the other

• May occur in special environments where each species consumes different

limiting substrates and neither species is affected by the end products of the
other. (e.g. The growth of yogurt starter strains of Streptococcus and
Lactobacillus in a chemostat.)

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MUTUALISM AND PROTOCOOPERATION
• In both cases, the presence of each population has a positive effect on the other.

• For mutualism, the interaction is essential to the survival of both species. (e.g.
aerobic bacteria and photosynthetic algae.)

• In protocooperation, the interaction is nonessential. One mechanism is the mutual

exchange of required substances or the removal of toxic end products by each
organism.

• An example is the growth of a phenylalanine-requiring strain of Lactobacillus and a

folic-acid-requiring strain of Streptococcus in a mixed culture.

• Very close mutualistic ties, such that partnership is necessary for the survival of one
or both species, are often termed symbiosis.
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 COMMENSALISM

• One population is positively affected by the presence of the other.

• The second population is not affected by the presence of the first population.

• Two common mechanisms are the following:

(a) The second population produces a required nutrient or growth factor for
the first population. (e.g. production of H2S by Desulfovibrio (through the
reduction of SO2− 4 ), which is used as an energy source by sulfur bacteria.)
(b) The second population removes a substance from the medium that is
toxic to the first population. (e.g. removal of lactic acid by the fungus
Geotrichium candidum, which allows the growth of Streptococcus lactis)
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 AMENSALISM

• Opposite of commensalism.

• Population A is negatively affected by the presence of the other population (B).

However, population B is not affected by the presence of population A.

• Two common mechanisms are the following:

(a) Population B produces a toxic substance that inhibits the growth of
population A. (e.g. production of antibiotics by certain molds to inhibit the
growth of others)
(b) Population B removes essential nutrients from the media, thus
negatively affecting the growth of population A.
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 PREDATION AND PARASITISM

• One population benefits at the expense of the other.

• Predation involves the ingestion of prey by the predator organism. (e.g. the
ingestion of bacteria by protozoa)

• In parasitism, the host, which is usually the larger organism, is damaged by

the parasite.

• The parasite benefits from utilization of nutrients from the host. (e.g. The
destruction of microorganisms by microphages.)

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16.3. SIMPLE MODELS DESCRIBING MIXED-
CULTURE INTERACTIONS

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EXAMPLE 16.1.
Competition of two species for the same growth-rate-limiting substrate is common.
Determine when the two organisms may stably coexist if both A and B follow Monod
kinetics.
Solution For this situation, the following equations describe the dynamic situation:

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16.4. MIXED CULTURES IN NATURE

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16.4. MIXED CULTURES IN NATURE

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16.5. INDUSTRIAL UTILIZATION OF MIXED CULTURES

• Defined mixed microbial populations are commonly used in cheese making,

a good example of using mixed cultures in food production.

• Streptococcus and Lactobacillus are the bacteria used for lactic acid
production in a mixed culture.

• Brevibacterium linens, Propionibacterium shermanii, Leuconostoc sp.,

and Streptococcus diacetilactis are the organisms are used to develop
flavor and aroma

• Penicillum camemberti and Penicillum roqueforti are some molds used in

cheese making.
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• The Symba process was developed in Sweden for treating starchy wastes,
particularly those from potato processing.

• Corn and pea wastes are also treated by a mixed culture of Trichoderma
viride and Geotrichium sp.

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 16.6. BIOLOGICAL WASTE TREATMENT:AN
EXAMPLE OF THE INDUSTRIAL UTILIZATION OF
MIXED CULTURES

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16.6.1. OVERVIEW

Three major Categories of Waste Materials:

• Industrial wastes are produced by various industries, and waste

characteristics vary greatly from one industry to another.

• Domestic wastes are treated by municipalities and derive from humans and
their daily activities. (e.g. ground garbage, laundry water, excrement, and often
some industrial wastes)

• Agricultural wastes are produced by farm animals (e.g., manure) and include
waste plants, such as straws.

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Three major waste treatment methods:

1. Physical treatment includes screening, flocculation, sedimentation, filtration,

and flotation, which are usually used for the removal of insoluble materials.

2. Chemical treatment includes chemical oxidations (chlorination, ozonation)

and chemical precipitation using CaCl2, FeCl3, Ca(OH)2, or Al2(SO4)3.

3. Biological treatment includes the aerobic and anaerobic treatment of waste

water by a mixed culture of microorganisms.

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Carbon content (strength) of a waste-water sample can be
expressed in several ways:

1. Biological oxygen demand (BOD)

2. Chemical oxygen demand (COD)
3. Total organic carbon (TOC)

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BOD (BIOLOGICAL OXYGEN DEMAND)

• The amount of oxygen required by microorganisms to break down organic

matter in a waste water.

• Normally, a 5-day BOD value is reported. The BOD5 is the amount of

dissolved oxygen (DO) consumed when a waste-water sample is seeded
with active bacteria and incubated at 20∞C for 5 days.

• BOD5 is calculated using the following equation:

This method is applicable only to biodegradable, soluble organics and requires a high concentration of active bacteria
preadapted to this type of waste. Moreover, if organic compounds are refractory, 5 days of incubation may not suffice,
and 20 days of incubation (BOD20) may be required.
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COD (CHEMICAL OXYGEN DEMAND)

• COD is a measure of the concentration of chemically oxidizable organic

compounds present in waste water.

• Organic compounds are oxidized by a strong chemical oxidant, and using the
reaction stoichiometry, the organic content is calculated.

• COD content of a waste-water sample usually exceeds the measured BOD

(COD > BOD5).

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TOC (TOTAL ORGANIC CARBON))

• The TOC content of waste-water samples can be determined by using a TOC

analyzer.

• After proper dilutions, samples are injected into a high-temperature (900° to

950°F) furnace and all organic carbon compounds are oxidized to CO2,
which is measured by an infrared analyzer.

• To determine the TOC content, waste-water samples should be acidified to

remove inorganic carbon compounds (mainly carbonates). The total carbon
content of waste water can be determined before and after acidification, and
the difference is inorganic carbon content.

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A typical waste-treatment operation employing biological treatment
includes the following steps:

1. Primary treatment includes the removal of coarse solids and suspended

matter (screening, sedimentation, filtration) and conditioning of the waste-water
stream by pH adjustment and nutrient additions (e.g., PO34 , NH+ 4).

2. Secondary treatment is the major step in biological treatment; it includes

biological oxidation or anaerobic treatment of soluble and insoluble organic
compounds.

3. Tertiary treatment includes the removal of the remaining inorganic compounds

(phosphate, sulfate, ammonium) and other refractory organic compounds by
one or more physical separation methods, such as carbon adsorption, deep-
bed filtration, and in some cases membrane-based techniques, such as
reverse osmosis or electrodialysis.
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16.6.2. BIOLOGICAL WASTE TREATMENT
PROCESSES

• Could be aerobic or anaerobic.

• The major aerobic processes (or reactor types) used

in waste-water treatment are (1) activated sludge, (2)
trickling filter, (3) rotating biological contractors, and
(4) oxidation ponds.

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 Activated-sludge processes include a well-agitated and aerated continuous-
flow reactor and a settling tank.
A long narrow tank with single feed approaches PFR behavior; circular tanks
approach CFSTR.

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THANK
YOU
Sherry Anne Y. Panganiban
PRESENTOR