GENERAL MICROBIOLOGY Chapter no 1

Introduction Definition: Microbiology is the study of living organisms of microscopic size,

which include bacteria, fungi, algae, protozoa, and the infectious agents at the borderline of life
that are called viruses. Pharmaceutical Microbiology Pharmaceutical Microbiology is an
applied branch of microbiology. It involves the study of: microorganisms concerned with the
manufacture of pharmaceuticals e.g. minimizing the number of microorganisms in a process
environment, excluding microorganisms and microbial bi-products like exotoxins and endotoxins
from water and other starting materials, and ensuring the finished pharmaceutical product is
sterile.
Microbiology as a field of biology Biologists having different opinions as how biology can best
be subdivided. o Divisions based on different groups of life:
❖Zoology (Animals) ❖Botany (Plants) ❖Entomology (Insects) ❖Microbiology
(Microorganisms) o Divisions based on the level at which study is conducted: ❖Molecular

form and functions: ❖Morphology ❖Physiology ❖Metabolism ❖Genetics ◦ Microbes or

biology ❖Cell biology ❖Organismal biology ❖Population biology o Divisions on the basis

◦ Microbiologists are concerned with the characteristics and functions such as morphology,
microorganisms are minute living things that are usually unable to be viewed with the naked eye.

physiology, taxonomy, genetics and molecular biology. ◦ Microorganisms are closely associated
with the health and welfare of human beings i.e. production of fermented products such as

and to decompose organic wastes. ◦ Harmful effects of microorganisms include deterioration of

vinegar, cheese and bread, production of industrial chemicals such as ethyl alcohol and acetone

microbiology: ◦ Historians are unsure who made the first observation of microorganisms but
materials like iron pipes, glasses and lenses and can cause disease and spoil food. History of

microscope was available during mid 1600s. ◦ An English scientist named Robert Hooke (1635 –

other plant materials. He observed strands of fungi among the specimens of cells he viewed. ◦
1703) made key observations (1665) of the fine structure of cork, honeycomb like structure and

The concept of cell theory is credited by two Germans, Matthias Schleiden and Theodor
Schwann (1838 - 1839) described cell as the basic structural and functional units of all

◦ Investigators described the Protoplasm (Greek proto, “first” and plasm, “formed substance”)
organisms. They recognized that cells, no matter what the organism, are very similar in structure.

introduced to characterize the living material of a cell is a colloidal organic complex consisting

contains nuclei or an equivalent nuclear substance. ◦ In 1670s a Dutch merchant named Anton
largely of protein, lipids, and organic acids enclosed by membranes or cell walls; always

Van Leeuwenhoek made careful observations of microscopic organisms which he called as

fungi and bacteria. ◦ After the death of Leeuwenhoek study of microbiology didn’t develop
animalcules, and he is regarded as one of the first to provide accurate description of protozoa,

because microscopes were rare. In those years theory of spontaneous generation was debated
which stated that microorganisms arise from lifeless matter such as beef broth. This theory was

decaying meat if meat is covered to prevent the entry of flies. ◦ Louis Pasteur worked in middle
disputed by Francesco Redi (1626 – 1697) who showed that fly maggots didn’t arise from

and late 1800s. He performed various experiments to discover why wine and dairy products
become sour and he found that bacteria were to blame. Pasteur had to disprove spontaneous
generation to sustain his theory and he devised a series of swan-necked flasks filled with broth.

in neck not the broth. Thus flasks didn’t become contaminated. ◦ Pasteur postulated the germ
He left the flasks open to air but the flasks had curve in neck so that microorganisms would fall

theory of disease which states that microorganisms are the cause of infectious disease. ◦ Pasteur
attempt to prove the germ theory were unsuccessful. However German scientist Robert Koch
provided the proof by cultivating Anthrax bacteria apart from any other type of organism. He

anthrax. This procedure came to be known as Koch’s postulates. ◦ Carolus Linnaeus (1707 –
then injected pure culture of bacilli into mice and showed that the bacilli invariably caused

basis the present day classification in biology was established. ◦ In the early eighteenth century,
1778), a Swedish botanist was given the concept of classification and nomenclature. On this

animal. ◦ In 1866, a German zoologist, E. H. Haeckel suggested a third kingdom, Protista. ◦ In

the classification of living organisms placed all organisms into one of two kingdoms, plant and

the late 1800s and the first decade of 1900s there emerged a golden age of microbiology during

microbial diseases were discovered during that period. ◦ After world war II antibiotics were
which many agents of infectious diseases were identified. Many of the etiologic agents of

use of antibiotics. ◦ In 1940s electron microscope was developed. In that decade cultivation
introduced to medicine. The incidence of pneumonia, T.B and meningitis were declined with the

methods for viruses were also introduced and knowledge of viruses developed rapidly. ◦ In 1952,

is produced by soil bacterium. ◦ In 1950s and 1960s viral diseases like polio, measles, mumps
Waksman was awarded the NOBEL PRIZE in the discovery of antibiotics Streptomycin, which

and rubella came under control. ◦ In 1969, R. H. Whittaker proposed more recent and
comprehensive classification, the five kingdom system. ◦ Modern microbiology reaches into
many fields including development of pharmaceutical products, use of quality control methods in
food and dairy products and industrial application of microorganism. Microorganisms also
produce vitamins, amino acids, enzymes and growth supplements. One of the major area of
applied microbiology is biotechnology where microorganisms are used as living factories to
produce pharmaceuticals e.g. insulin, blood clotting factors and number of vaccines.
1. Classification of microorganisms ◦ Microorganisms differ enormously in terms of their
shape, size and appearance and in their genetic and metabolic characteristics. All these properties
are used in classifying microorganisms into the major groups with which many people are

as chlamydia, rickettsia and mycoplasmas. ◦ 1.1) viruses, viroids and prions: ◦ Viruses do not
familiar, e.g. bacteria, fungi, protozoa and viruses, and into the less well known categories such

have a cellular structure. They are particles composed of nucleic acid surrounded by protein;
some possess a lipid envelope and associated glycoproteins, but recognizable chromosomes,
cytoplasm and cell membranes are absent. Viruses are incapable of independent replication as
they do not contain the enzymes necessary to copy their own nucleic acids; as a consequence, all
viruses are intracellular parasites and are reproduced using the metabolic capabilities of the host
cell. A great deal of variation is observed in shape (helical, linear or spherical), size (20–400 nm)
and nucleic acid composition double stranded (single- or , linear or circular RNA or DNA), but

microscope; instead they may be viewed using an electron microscope. ◦ Viroids (virusoids) are
almost all viruses are smaller than bacteria and they cannot be seen with a normal light
even simpler than viruses, being infectious particles comprising single-stranded RNA without

no known human pathogens in this category. ◦ Prions are unique as infectious agents that they
any associated protein. Those that have been described are plant pathogens, and, so far, there are

contain no nucleic acid. A prion is an atypical form of a mammalian protein that can interact
with a normal protein molecule and cause it to undergo a conformational change so that it, in
turn, becomes a prion and ceases its normal function. Prions are the agents responsible for

spongiform encephalopathy (BSE) or mad cow disease. ◦

transmissible spongiform encephalopathies, e.g. Creutzfeldt– Jakob disease (CJD) and bovine

1.2) prokaryotes and eukaryotes: ◦ The most fundamental distinction between the various
microorganisms having a cellular structure (i.e. all except those described in section 1.1 above) is

cellular structure and mode of reproduction. ◦ prokaryotes are the bacteria and archaea and
their classification into two groups- prokaryotes and eukaryotes— based primarily on their

eukaryotes are all other cellular microorganisms, e.g. fungi, protozoa and algae. ◦ prokaryotes
are normally haploid (possess only one copy of the set of genes in the cell) and reproduce

normally have the potential to reproduce sexually. ◦ The major difference between these two
asexually; eukaryotes, by contrast, are usually diploid (possess two copies of their genes) and

types of cell is the possession by the eukaryotes of a true cell nucleus in which the chromosomes
are separated from the cytoplasm by a nuclear membrane. The prokaryotes have no true nucleus;
they normally possess just a single chromosome that is not separated from the other cell contents
by a membrane. 1.2.1
)Bacteria and archaea: ◦ Bacteria are essentially unicellular, although some species arise as
sheathed chains of cells. ◦ The bacteria of interest in pharmacy and medicine belong to the group
known as the eubacteria. The other subdivision of prokaryotes, the archaea, have little or no
pharmaceutical importance and largely comprise organisms capable of living in extreme
environments (e.g. high temperatures, extreme salinity or pH) or organisms exhibiting

production of methane). ◦ The eubacteria are typically rod-shaped (bacillus), spherical (cocci),
specialized modes of metabolism (e.g. by deriving energy from Sulphur or iron oxidation or the

curved or spiral cells of approximately 0.5–5.0μm (longest dimension) and are divided into two

procedure developed in 1884 by Christian Gram . ◦ Some of the bacteria that contaminate or
groups designated Gram-positive and Gram negative according to their reaction to a staining

cause spoilage of pharmaceutical materials are saprophytes, i.e. they obtain their energy by
decomposition of animal and vegetable material, while many could also be described as parasites

pathogens (parasites damaging the host). ◦ Rickettsia and chlamydia are types of bacteria that are
(benefiting from growth on or in other living organisms without causing detrimental effects) or

cannot easily be cultivated in the laboratory. ◦ Most of the bacteria grow well at temperatures
obligate intracellular parasites, i.e. they are incapable of growing outside a host cell and so

between ambient and human body temperature, and exhibit wide variations in their requirement
for, or tolerance of, oxygen. Strict aerobes require atmospheric oxygen, but for strict anaerobes
oxygen is toxic. Many other bacteria would be described as facultative anaerobes (normally

concentrations lower than those in normal air). ◦ 1.2.2) Fungi : ◦ Fungi are eukaryotes and
growing best in air but can grow without it) or micro- aerophils (preferring oxygen
structurally more complex than bacteria. Fungi are considered to be non photosynthesizing

these two groups is not always clear. ◦ Yeasts are normally unicellular organisms that are larger
plants, and the term fungus covers both yeasts and moulds, although the distinction between

than bacteria (typically 5–10μm) and divide either by a process of binary fission or budding

independent existence) ◦ Mould is an imprecise term used to describe fungi that do not form
(whereby a daughter cell arises as a swelling from the parent that eventually separates to lead an

filaments or threads (hyphae) which vary between 1 and > 50mm wide. ◦ Although fungi are
fruiting bodies visible to the naked eye. Most moulds consist of a tangled mass (mycelium) of

which this has never been observed. ◦ Most fungi are saprophytes with relatively few having
eukaryotes that should, in theory, be capable of sexual reproduction, there are some species in

pathogenic potential. ◦ 1.2.3) Protozoa: ◦ Protozoa are eukaryotic, predominantly unicellular

microorganisms that are regarded as animals rather than plants. ◦ Many protozoa are free-living

including humans, e.g. the organisms responsible for malaria and amoebic dysentery. ◦ 1.2.4)
motile organisms that occur in water and soil, although some are parasites of plants and animals,

Algae: ◦ Algae are eukaryotes which can exist as single cells, an example of which is

instance Rhodymenia (red seaweed). ◦ All algae contain a pigment called chlorophyll a (other
Chlamydomonas, or joined together in chains like Spirogyra or made up of many cells, for

types of chlorophyll such as b, c and / or d may also be present) and they make their own food by

appearance. 2) Naming of microorganisms: ◦ Aristotle classified all living things as plants or

photosynthesis. The chlorophyll is contained in the chloroplasts and gives many algae their green

animals. ◦ Swedish botanist Carl Linnaeus made great efforts in classification of living things i.e.
Binomial nomenclature. ◦ Microorganisms, just like other organisms, are known by two names:
that of the genus (plural=genera) and that of the species. ◦ The former is normally written with

aureus or Escherichia coli. ◦ These may be abbreviated by shortening the name of the genus
an upper case initial letter and the latter with a lower case initial letter, e.g. Staphylococcus

provided that the shortened form is unambiguous, e.g. staph. aureus, [Link]. ◦ Both the full and
the shortened names are printed in italics to designate their status as proper names. ◦ The species
within a genus are sometimes referred to by a collective name, e.g. staphylococci or
pseudomonads, and neither these names, nor names describing groups of organisms from
different genera, e.g. coliforms, are italicized or spelt with an upper case initial letter.
Concept Scope of microbiology with reference to pharmaceutical science: ◦ Pharmaceutical
microbiology: ◦ Pharmaceutical microbiology is the applied branch of microbiology which
allow pharmacist to manufacture pharmaceuticals from microorganisms either directly or with
the use of some products produced by them. Other aspects of pharmaceutical microbiology

agents. ◦
includes research and development for manufacturing of various anti tumors and anti microbial

Scope of microbiology: ◦ Criteria and standards for the microbiological quality of medicines
depend upon the route of administration of the medicine in question. The vast majority of
medicines that are given by mouth or placed on the skin are non-sterile, i.e. they may contain
some microorganisms (within limits on type and concentration), whereas all injections and
ophthalmic products must be sterile, i.e. they contain no living organisms. For a sterile product
the criterion of quality is simple; there should be no detectable microorganisms whatsoever. The
product should, therefore, be able to pass a test for sterility, and a knowledge of the procedures

◦ Injections are also subject to a test for pyrogens; these are substances that cause a rise in body
and interpretation of results of such tests is an important aspect of pharmaceutical microbiology.

temperature when introduced into the body. Strictly speaking, any substance which causes fever
following injection is a pyrogen, but in reality the vast majority are of bacterial origin, and it is

considered within the realm of microbiology. ◦ Sterile medicines may be manufactured by two
for this reason that the detection, assay and removal of bacterial pyrogens (endotoxins) are

different strategies. The most preferred option is to make the product, pack it in its final
container and sterilize it by heat, radiation or other means. The alternative is to manufacture the
product from sterile ingredients under conditions that do not permit the entry of contaminating
organisms. Those responsible for the manufacture of sterile products must be familiar with the
sterilization or aseptic manufacturing procedures available for different product types, and those
who have cause to open, use or dispense sterile products (in a hospital pharmacy, for example)

product contamination. ◦ There is a large range of antimicrobial drugs used to prevent and treat
should be aware of the aseptic handling procedures to be adopted in order to minimize the risk of

microbial infections. Because of this range and diversity of products, pharmacists are now far

particular categories of infection. ◦ Microorganisms are valuable in the maintenance of our

more commonly called upon to advise on the relative merits of the antibiotics available to treat

plant and animal material and in the fixation of atmospheric nitrogen. ◦ The uses of
ecosystems. Their role and benefits in the carbon and nitrogen cycles in terms of recycling dead

microorganisms in the food, dairy and brewing industries are also well established. Applications
of Microbiology to food fall into 2 general categories: 1)Preserving food from microbes, making
sure that harmful microbes or toxins produced by them are not in the food 2)Using microbes to
produce particular types of food such as cheese (bacteria), beer (yeast), bread (yeast), yogurt
(bacteria), etc. oApart from these major applications, however, the uses of microorganisms in the
manufacture of medicines prior to 1980 were very limited. Enzymes were developed for use in
cancer chemotherapy (asparaginase) and to digest blood clots (streptokinase), and
polysaccharides also found therapeutical applications (e.g. dextran— used as a plasma
expander). oAnother major advancement of microbiology is recombinant DNA technology in the
1970s. This technology permitted human genes to be inserted into microorganisms, which were
thus able to manufacture the gene products far more efficiently than traditional methods of
extraction from animal or human tissues. Insulin, in 1982, was the first therapeutic product of
DNA technology to be licensed for human use, and it has been followed by human growth

permitted the development of vaccines e.g. hepatitis B vaccine. ◦ All these developments,
hormone, interferon, blood clotting factors and many other products. DNA technology has also

together with miscellaneous applications in the detection of mutagenic and carcinogenic activity
in drugs and chemicals and in the assay of antibiotics, vitamins and amino acids have ensured
that the role of microorganisms in the manufacture of medicines is now well recognized, and that
a basic knowledge of immunology , gene cloning is an integral part of pharmaceutical
microbiology.
Pharmaceutical importance of the major categories of microorganisms: ◦
Virus: ◦ The importance of viruses derives exclusively from their pathogenic potential. Because
of their lack of intrinsic metabolism viruses are not susceptible to antibiotics, and the number of
effective synthetic antiviral drugs is limited. For these reasons, viral infections are among the
most serious and difficult to cure, and of all the categories of microorganism, only viruses appear

Pathogens. ◦ Because they are not free-living, viruses are incapable of growing on manufactured
in (the most serious) Hazard Category 4 as classified by the Advisory Committee on Dangerous

medicines or raw materials, so they do not cause product spoilage. ◦ Viruses are relatively easy

perspective. ◦
to destroy by heat, radiation or toxic chemicals, so they do not represent a problem from this

Bacteria: ◦ Many of the medically and pharmaceutically important bacteria are pathogens, and

activity of antibiotics and biocides (disinfectants, antiseptics and preservatives). ◦ A wide variety
some of these pathogens are of long-standing notoriety as a result of their ability to resist the

enzymes and carbohydrates. ◦ The ability of bacteria to grow on diverse substrates ensures that
of bacteria are exploited commercially in the manufacture of other medicines including steroids,

their potential as agents of spoilage in manufactured medicines and raw materials is well
recognized, and the ability of many species to survive drying means that they survive well in
dust and so become important as contaminants of manufactured medicines. The ability to survive

well exemplified by bacterial spores. ◦

not only in dry conditions but in other adverse environments (heat, radiation, toxic chemicals) is

Fungi: ◦ Fungi are able to form spores that survive drying, so they too arise commonly as
contaminants of manufactured medicines. However, the degree of resistance presented by the

problem. ◦ Fungi do not generally create a significant infection hazard ; relatively few fungal
spores is usually less than that exhibited by bacteria, and fungi do not represent a sterilization

species are considered major pathogens for animals that possess a fully functional immune
system. There are several fungi which, while representing little threat to immunocompetent
individuals, are nevertheless capable of initiating an infection in persons with impaired immune

possessing this characteristic. ◦

function; the term opportunist pathogens is used to describe microorganisms (of all types)

Protozoa:
◦ Protozoa are of significance due to the pathogenic potential of a few species. Because protozoa

manufacturing environment. ◦ Protozoal infections are not currently a major problem to human
do not possess cell walls they do not survive drying well, so they are not a problem in the

health in temperate climates, although they are more troublesome in the tropics.