Microbiology

History
Introduction
Classification

MRS. E. MUONGA
 What is Microbiology

 Microbiology is the study of microorganisms.

 It is a branch of biology that deals with the investigation of microscopic organisms, also known as microbes,
that are either unicellular or multicellular in nature.
 These microbes are found in the living bodies of plants and animals and also in the air, water, soil and marine
kingdom.
 These microbes includes bacteria, viruses, parasites.
 Microorganisms are microscopic in the sense that they are too small to be seen by the naked eye.
 History of Microbiology and
Invention of the microscope
 The history of microbiology begins with the invention of the microscope.
 Anton Van Leeuwenhoek is called the “inventor of the microscope”
 He was a Dutch draper whose hobby was lens grinding and making microscopes.
 He created a “simple” microscope that could magnify to about 275x, and published drawings of
microorganisms in 1683
 He used simple ground lenses to reach magnifications of over 200x
 Robert Hooke developed the microscope further into what is now called the compound microscope
 The microscope has improved greatly over the years and now has
(i) An objective lens at the bottom that gathers light, magnifies and focuses the image inside the
body tube.
(ii) A body tube that helps with focusing the image and
(iii) An ocular lens at the top that magnifies the image again
 The Discovery of Microorganisms
 Almost 100 years later, the Danish biologist Otto Müller extended van Leeuwenhoek’s studies and organized bacteria
into genera and species according to the classification methods of Carolus Linnaeus.
 This was the beginning of the taxonomic classification of microbes.
 In 1840, the German pathologist Friedrich Henle proposed criteria for proving that microorganisms were responsible
for causing human disease (the “germ theory” of disease).
 Robert Koch and Louis Pasteur confirmed this theory in the 1870s and 1880s with a series of elegant experiments
proving that microorganisms were responsible for causing anthrax, rabies, plague, cholera, and tuberculosis.
 Other brilliant scientists went on to prove that a diverse collection of microbes was responsible for causing human
disease.
 The era of chemotherapy began in 1910, when the German chemist Paul Ehrlich discovered the first antibacterial
agent, a compound effective against the spirochete that causes syphilis. This was followed by Alexander Fleming’s
discovery of penicillin in 1928, Gerhard Domagk’s discovery of sulfanilamide in 1935, and Selman Waksman’s
discovery of streptomycin in 1943. In 1946, the American microbiologist John Enders was the first to cultivate viruses
in cell cultures, leading the way to the large-scale production of virus cultures for vaccine development.
 Thousands of scientists have followed these pioneers, each building on the foundation established by his or her
predecessors, and each adding an observation that expanded our understanding of microbes and their role in disease.
 Microorganisms and Diseases
Koch’s Postulates
 In 1884, Koch published four postulates that summarized his method for determining whether a
particular microorganism was the cause of a particular disease.
 Each of Koch’s postulates represents a criterion that must be met before a disease can be
positively linked with a pathogen.
 In order to determine whether the criteria are met, tests are performed on laboratory animals and
cultures from healthy and diseased animals are compared.
Koch’s Postulates
1. The suspected pathogen must be found in every case of disease and not be found in healthy individuals.
2. The suspected pathogen can be isolated and grown in pure culture.
3. A healthy test subject infected with the suspected pathogen must develop the same signs and symptoms of
disease as seen in postulate 1.
4. The pathogen must be re-isolated from the new host and must be identical to the pathogen from postulate 2.
What are Microorganisms

 There are several classes of living things.

 Based on the organization of their cellular structures, living organisms can be
Eukaryotes or Prokaryotes.
 Eukaryotes are much larger and more complex than prokaryotes. Their processes
are compartmentalized into organelles.
 Prokaryotes represent 2 domains i.e bacteria and archaea. Archaea usually live in
Earth’s extreme environments while bacteria are the most abundant and
diversified organisms on Earth. However, they lack membrane-bound organelles
and their processes are not compartmentalized.
Microorganisms continued…

 Microbes (microorganisms) have both specific and special characteristic features

that enable them to survive adequately in a wide spectrum of environments.
 They can survive in streams, ponds, lakes, rivers, oceans, ice, water-borne pipes,
hot springs, gastro-intestinal tracts, roots of plants and even in oil wells.
 Microbiology is therefore the study of tiny organisms that cannot be seen by the
naked eye and also includes the study of the microorganisms’ distribution in
nature, relationship to each other and other living microorganisms, their effects on
human health and their reaction to various physical and chemical agents.
Different fields of medical microbiology

 Microbiology can also be divided into 2 broad categories namely Industrial and Medical microbiology.
Industrial Microbiology
 This is a branch of microbiology that deals with the study and use of various microorganisms that are responsible for
the production of many products such as alcoholic products, antibiotics, enzymes and even vaccines.
 The use of genetically modified microorganisms (GMOs) is also another advanced field of microbiology.
a. Industrial microbiology includes the following fields:
(i) Aero-Microbiology- concerned with the preservation and preparation of food, food-borne illnesses and their
ultimate prevention
(ii) Beverage Microbiology-concerned with the making of beer, shandy, wine, and a variety of alcoholic beverages
e.g whisky, brandy, rum, gin, vodka, etc.
(iii) Exomicrobiology - concerned with exploration of life in outer space.
(iv) Food microbiology-concerned with the making of cheese and yoghurt.
(v) Geochemical microbiology-concerned with the study of coal, mineral deposits, gas formation, prospecting the
deposits of gas and oil and recovery of minerals from low-grade ores.
(vi) Waste-treatment microbiology-concerned with treatment of domestic and industrial effluents/waste.
Fields of microbiology continued…

b. Medical Microbiology
 Concerned with making diagnostic protocols for identification of causative agents
of various human ailments and subsequent preventive measures.
 The clinical microbiology laboratory plays an important role in the diagnosis and
control of infectious diseases.
 However, the ability of the laboratory to perform these functions is limited by the
quality of the specimen collected from the patient, the means by which it is
transported from the patient to the laboratory, and the techniques used to
demonstrate the microbe in the sample.
 Because most diagnostic tests are based on the ability of the organism to grow,
transport conditions must ensure the viability of the pathogen.
 Importance of Microbiology

1. Important in the making of life-saving drugs or antibiotics such as penicillin, ampicillin, chloramphenicol,
ciprofloxacin, tetracycline, streptomycin.
 Some of these life-saving drugs are derived from microorganisms.
2. Microorganisms are protectors of the environment, ensuring that minerals e.g. carbon and nitrogen are
recycled
3. In combating infectious diseases
4. Useful in the food industry where they can be used as active cultures
Different fields of microbiology

 The following are the different fields of microbiology depending on the organism
of interest.
 Bacteriology- the study of bacteria
 Mycology-the study of fungi
 Phycology-the study of algae
 Protozoology-the study of protozoans
 Virology-the study of viruses
 Parasitology-the study of parasites (which includes all the above organisms)
Classification of
Bacteria
Ways of classifying bacteria

 Various classifications have been brought forth but the following are widely
accepted:
1. According to morphology – their physical characteristics under the microscope
2. According to physiology – their enzymatic and biochemical characteristics
3. According to their motility
4. According to gaseous requirements
5. According to nutritional requirements
6. According to temperature requirements
7. According to pH requirements
8. According to their cell wall composition
 Classification of Bacteria based on shape
(Morphology)
 Depending on their shape, bacteria are classified into several varieties
1. Cocci (from kokkos meaning berry) are spherical or oval cells
2. Bacilli (from baculus meaning rod) are rod shaped cells
3. Vibrios are comma shaped curved rods and derive their name from their characteristic vibratory motility.
4. Spirilla are rigid spiral forms.
5. Spirochetes (from speira meaning coil and chaite meaning hair) are flexuous spiral forms
6. Mycoplasmas are bacteria that are cell wall deficient and hence do not possess a stable morphology. They
occur as round or oval bodies and as interlacing filaments.
 Classification of bacteria based on
Enzymatic and Biochemical Characteristics

Catalase Test
 Catalase is an enzyme produced by many bacteria
 It splits hydrogen peroxide into water and oxygen
 Hydrogen peroxide is a by-product of aerobic respiration and is lethal if it accumulates in the
bacterial cell
 Catalase enzyme degrades this hydrogen peroxide before it can do any damage to the bacterial cell
2H2O22H2O + O2
 The test is used to differentiate members of the family micrococci from streptococci
 It is also carried out for Mycobacteria to differentiate tubercle bacilli from atypical mycobacteria
 Streptococci are catalase negative while micrococci are catalase positive
Oxidase Test

 The enzyme oxidase plays a vital role in the operation of the electron transport
system during aerobic respiration
 Aerobic bacteria, as well as some facultative anaerobes and microaerophiles
exhibit oxidase activity.
 Oxidase positive bacteria include Vibrio spp,Pasteurella spp
 Oxidase negative bacteria include all members of the family enterobacteria
Coagulase Test

 The enzyme coagulase produced by a few Staphylococcus species is a key feature

of pathogenic Staphyloccocci
 The enzyme causes coagulation of blood, causing the organism to block off its
infection from the host’s immune system
 Coagulase is able to convert fibrinogen into fibrin which results in the formation
of a visible clot
 The test is used to identify S. aureus and differentiate it from other species of
Staphylococcus
Urease Test

 This enzyme is possessed by certain fungi and bacteria

 It hydrolyses urea releasing ammonia into the medium
 This causes a change in pH which is detected by the colour change in the indicator
dye
 It can be used to differentiate groups of bacteria and fungi
Indole Test

 Tryptophan is an essential amino acid that can undergo oxidation by enzymatic

activities of some bacteria
 Tryptophan is converted into metabolic products by the enzyme tryptophanase
 Tryptophanase converts tryptophan into the metabolic products indole, skatole
and indole acetic acid
 The ability to hydrolyse tryptophan into indole is not possessed by all bacteria.
 Examples of Indole positive bacteria include E. coli and Vibrio cholera
 Examples of Indole negative bacteria are Klebsiella pneumoniae and Salmonella
typhi
Methyl Red Test

 All enteric bacteria ferment glucose with the production of organic acids and
energy
 The Methyl red test detects the presence of high concentrations of acidic end
products
 Examples of MR positive bacteria are E. coli and Salmonella
 Examples of MR negative bacteria are Klebsiella pneumoniae and Enterobacter
species
Voges-Proskauer Test

 Named after 2 microbiologists

 The test determines the capability of some bacteria to produce neutral end-
products after glucose metabolism
 Most enteric bacteria produce acetoin as the main end product of glucose
metabolism and form smaller quantities of mixed acids
 Most enteric bacteria are VP negative with the exception of Klebsiella, Serratia
and a few other organisms
Citrate Utilization Test

 Some bacteria can also utilize citrate as their sole source of carbon
 In the absence of fermentable glucose or lactose, some bacteria are able to utilize
citrate as a sole source of carbon for their energy.
 Sodium citrate is useful in the tricarboxylic acid cycle
 Examples of citrate positive bacteria are Klebsiella and Citrobacter
 Examples of citrate negative bacteria are E. coli and Salmonella typhi
Classification of bacteria based on motility

 Bacteria can be motile or non-motile based on the presence of structures called

flagella that enable them to move around
 These movements of motile bacterial cells depend on the arrangement of the
flagella around the bacterial cell
 Therefore, bacteria can be said to be:
(i) Monotrichous – single polar flagellum located on one end
(ii) Amphitrichous – With two flagella, one on each end
(iii) Lophotrichous – With two or more flagella at one or both ends
(iv) Peritrichous – Flagella all over the surface
(v) Atrichous – Bacteria without flagella (e.g. cocci which rarely have flagella)
Flagella arrangements
 Classification based on gaseous
requirements
 Depending on the influence of oxygen on growth and viability, bacteria are
divided into aerobes and anaerobes.
 Aerobic bacteria require oxygen for growth.
 They may be like Vibrio cholera, which will grow only in the presence of
oxygen or like other organisms which are ordinarily aerobic but can grow
in the absence of oxygen.
 Most bacterial of medical importance fall into the latter category (are
ordinarily aerobic but can grow in the absence of oxygen).
 Bacteria such as Clostridia, grow in the absence of oxygen and may even
die on exposure to oxygen.
 Another group of bacteria grow best in the presence of low oxygen.
Classification based on gaseous
requirements
Obligate aerobes
- Require oxygen to grow, cannot grow in environments with no oxygen
- E.g pseudomonas found commonly causing nosocomial infections
Facultative anaerobes
- Can use oxygen but can grow in its absence
- E.g staphylococcus spp.
Obligate Anaerobes
Cannot use oxygen, harmed by the presence of oxygen
Grow well in environments that don’t have oxygen
e.g Clostridium spp.
Microaerophilic
- Require oxygen, but at low concentrations e.g Brucella spp.
Classification based on pH

 Most bacteria prefer neutral pH (6.5 – 7.5)

 Acidity inhibits microbial growth and is used as a preservation
technique (e.g pickles)
 Alkalinity also inhibits microbial growth, but is not commonly used
for preservation purposes
 However, some organisms are able to grow in both acidic and alkaline
environments
 Acidic –acidophiles – (pH of 0.1-5.4) – e.g lactobacillus sp.
 Alkaline- alkalophiles (pH greater than 8.5) e.g Vibrio cholerae
 Neutral pH-neutrophiles – (pH 5.4 to 8.5) – most bacteria
 Classification based on temperature
requirements
1. Psychrophiles – ‘cold-loving’ bacteria. These can further be divided into 2 groups.
(a) True psychrophiles – grow well at temperatures less than 15 degrees
- Optimum growth is at 15 degrees
- Usually don’t cause food spoilage
- Examples are bacteria that grow in cold places like the north pole, ocean depths
(b) Psychrotrophs – their optimum growth is at temperatures between 15 and 25 degrees Celsius
- Responsible for most low temperature food spoilage
2. Mesophiles – ‘middle loving’
- Most bacteria responsible for food spoilage are under this category
- Includes most pathogens and food spoilage organisms
- Most are adapted to live in the bodies of animals
- Optimum temperature is 37 degrees but grow well at temperatures between 25 and 40 degrees
 Classification based on temperature
requirements
3. Thermophiles – ‘heat loving’ bacteria.
- Grow at temperatures higher than 45 degrees
- Optimum growth is between 50 and 60 degrees
4. Extreme Thermophiles
- Also known as hyperthermophiles
- Grow well at temperatures higher than 80 degrees
- Most live in volcanic and ocean vents
 Classification of bacteria based on
nutritional requirements
 Bacteria can be classified nutritionally, based on their energy requirement
and on their ability to synthesize essential metabolites.
 Bacteria which derive their energy from sunlight are called phototrophs,
those who obtain energy from chemical reactions are called chemotrophs.
 Bacteria which can synthesize all their organic compounds are called
autotrophs and those that are unable to synthesise their own metabolites
are heterotrophs.
 Some bacteria require certain organic compounds in minute quantities.
These are know as growth factors or bacterial vitamins. Growth factors
are called essential when growth does not occur in their absence, or they
are necessary for it.
Classification of bacteria based on cell
wall composition
 Bacteria can be classified into two main groups based on the cell wall composition.
Though there is a group of bacteria that lack cell walls (The Mycoplasmas-acid fast
bacteria), those that have cell walls can broadly be classified as Gram positive and Gram
negative.
 The structure, chemical composition and thickness of the cell wall differs in gram positive
and negative bacteria.
1. The peptidoglycan layer is much thicker in gram positive than gram negative bacteria.
Some gram positive bacteria also have some fibers of teichoic acid that protrude outside
the peptidoglycan whereas the gram negative bacteria do not.
2. Gram Negative organisms have a complex outer layer consisting of lipopolysaccharide,
lipoprotein and phospholipid. Lying between the outer membrane and cytoplasmic
membrane in gram negative bacteria lies a periplasmic space which is the site where
some species have enzymes called beta-lactamases that degrade penicillins and other
beta-lactam drugs.