Bacteria live in a variety of habitats such as the soil, the ocean, and even the insides of the human gut.
They exist either as free-living organisms or as parasites. These are most successful living forms as they live in the diverse environment.
Bacterial cells are smaller and simpler than eukaryotic cells. They are an exceedingly diverse group of organisms that differ in size, shape, habitat, and even in metabolism.
This course covers classification of bacteria, gram staining, and bacterial reproduction.
Classification of Bacteria Based on the Morphology
Based on their body-type (morphology), bacteria can be classified into six distinct groups. Cocci and bacilli are the most common ones. These groups are described below.
These are oval or spherical shaped bacteria (Figure 2). They are arranged in diverse ways. Cocci can be classified on the basis of arrangement.
- Diplococcus: coccus in pairs. E.g, Neissseria gonorrhoae, Pneumococcus
- Streptococcus: coccus in chains or rows. Eg. Streptococcus salivarius
- Staphylococcus: coccus in irregular (grape-like) bunch. Eg. Streptococcus salivarius. Staphylococcus infections are commonly seen in food poisoning.
- Tetrad: Coccus in group of four. E.g. Micrococcus luteus, found in dust, soil, air and the skin of mammals.
- Sarcina: coccus in cubical arrangement of cell. E.g. Sporosarcina
These are rod-shaped bacteria. E.coli is the most commonly found bacilli present in human body. Based on arrangement, bacilli are further classified as shown below:
- Coccobacilli: E.g. Brucella
- Streptobacilli: chain of rod shape bacteria: E.g. Bacillus subtilis
- Comma shaped bacilli: E.g. Vibrio cholerae
They are bacteria that lack a cell wall. Mycoplasma is known as PPLO (Pleuropneumonia like organism).
It can cause sore throat, bronchitis, and pneumonia. These are smallest free-living organisms. E.g. Mycoplasma pneumoniae (Figure 4).
These are spiral-shaped bacteria. Spirochetes are a group comprised of six genera of bacteria. Examples of Spirochaetes are Spirochaeta, Treponema, Borrelia, and Leptospira.
They are causative agents of diseases like, syphilis and Lyme disease.
Rickettsiae and Chlamydiae
Rickettsiae and Chlamydiae (Figure 6) are gram negative bacteria and obligate intracellular parasite. They use host adenosine triphosphate (ATP), a source of energy, for their survival.
Both cannot be cultured on non-living artificial media; instead they are inoculated into living cells for culture. They are sensitive to antibiotics.
Actinomycetes are heterogeneous group of gram-positive, generally anaerobic bacteria (Figure 7).
These are one of the most diverse groups of filamentous bacteria. Examples of Actinomycetes are: Nocardia asteroids, Dermatophilus
Classification of Bacteria based on the Optimum pH for Growth
Based on the optimum acidity (pH) required for their growth, bacteria can be of three types:- acidophiles, alkaliphiles, and neutriphiles.
These types are described below.
- Bacteria that have grow optimally between pH 0 and 5.5 are called acidophiles.
- The cytoplasm of these bacteria is acidic in nature.
- Some acidopiles are thermophilic in nature. Such bacteria are called thermoacidophiles.
- Examples: Thiobacillus ferroxidans, Helicobacter pylori (Figure 8), Thermoplasma, Sulfolobus, Acidianus, Oligotropha corboxydovorans
- Bacteria that grow best in an alkaline environment (pH 9-11) are called alkaliphiles.
- Example: Vibrio cholerae, bacteria belonging to the genera Bacillus, Micrococcus, Pseudomonas, and Streptomyces
- Bacteria that grow best in a neutral environment (pH 6.5-7.5) are called neutrophiles.
- Most of the bacteria live in soil or water and grow at neutral pH.
- Example: E. coli (Figure 10)
Classification of Bacteria based on the Structure
A few different criteria are used to classify bacteria. They can be distinguished by the nature of their cell walls, by their shape, or by differences in their genetic makeup.
The structure of a bacterium is very simple as compared to the eukaryotic cells (Figure 11). It consists of following components discussed below.
Cell wall is found outside of the cell membrane. Peptidogylcan and Techoic acid are major components of the cell wall. Peptidoglycan is a porous cross-linked polymer that provides strength to cell wall.
Teichoic acid is water soluble polymer of glycerol or ribitol. Phosphate is present in gram positive bacteria. It is the major surface antigen.
Bacterial cells are surrounded by a variety of surface structures that allow them to survive in extreme environments.
The major function of the cell wall is to provide rigidity, strength, structural support, protection against mechanical stress, and infection. It also aids in osmotic regulation and diffusion of gases in and out of the cell.
In 1884, Dr. Christian Gram developed “Gram stain”. This stain helps to distinguish between Gram-positive and Gram-negative bacteria.
It is based on differential staining with a crystal violet-iodine complex and a safranin counterstain.
Gram stain is the most important and widely used microbiological differential staining technique. It also allows determination of cell morphology, size, and arrangement.
When stained with gram stain and fixed by a mordant, some bacteria are able to retain the stain by resisting decolorization after washing with decolorizer (acetone and absolute alcohol), while others get decolorized.
The bacteria that retain the primary stain are called Gram positive and those that get decolorized and then counterstained are called Gram negative (Figure 12).
The table below differentiates the gram-positive and gram-negative bacteria.
|Reaction to Gram Stain||Stain red or pink, they don’t retain the Gram stain when washed with absolute alcohol and acetone||Stain dark violet or purple, they retain the color even after washing|
|Peptidoglycan layer||Single layered and thin|
Multilayered and thick
|Lipid and lipoprotein content||High (due to presence of outer membrane)||Low|
|Flagellar structure||Four rings in basal body||Two rings in basal body|
|Toxin production||Produces endotoxins|
|Cell wall composition||Cell wall is 70-120A0 thick and two layered. Lipid content is 20-30%, Murein content is 10-20%||Cell wall is 100-120 A0 thick and single layered. Lipid content of the cell wall is low, whereas Murein content is 70-80%|
|Antibiotic resistance||Resistant to antibiotics|
Susceptible to antibiotics
|S layer||S layer is attached to outer membrane||S layer is attached to the peptidogycan|
|Resistance to sodium azide solution||High||Low|
|Pathogenecity||Most of them are pathogenic to humans and other animals||Very few forms are pathogenic to humans and other animals|
|Examples||Escherichia coli, Rhizobium, Vibrio, Acetobacter||Streptococcus, Clostridium, Lactobacillus, Bacillus subtilis|
Along with their differences, gram-positive and gram-negative bacteria have several similarities, as enumerated below:
- They both are prokaryotic, bacterial cells and lack membrane-bound organelles.
- They contain plasmid DNA along with circular DNA.
- Their cell wall is made of peptidoglycan and cytoplasm is surrounded by lipid bilayer with several proteins spanning it.
- They have both flagellated and non-flagellated forms.
- They undergo transformation, conjugation, and transduction
Bacterial cell membrane is also referred as cytoplasmic or inner membrane. It lies internal to the cell wall and encloses cytoplasm of the bacterium. This membrane is a functional equivalent of eukaryotic plasma membrane.
This membrane is a fluid phospholipid bilayer embedded with proteins. The phospholipid bilayer is arranged so that the polar ends of the molecules form the outermost and the innermost surfaces of the membrane.
The non-polar ends form the center of the membrane. Functions of cell membrane are listed below:
- Selective permeability
- Energy production
- Removal of waste
- Endospore formation
Glycocalyx helps in adhesion, protection from desiccation, and attack by a host’s immune system.
Capsule is present in capsulated bacteria (Baciilus subtilis, Haemplhilus influenza, Streptococcus pneumoniae, Klebsiella pneumoniae etc).
Most prokaryotes contain a polysaccharide layer outside of the cell wall. This layer is called capsule.
This capsule is 0.2µm thick layer firmly attached to the cell wall of the capsulated bacteria.
In Bacillus anthracis, it is made up of polypeptide (a polymer of D-glutamic acid) and in Streptococci, it is made of L-aminoacids.
Functions of capsule are as follows:
- Virulence factor: capsule enhances the ability of bacteria to cause disease by preventing their phagocytosis.
- Capsule prevents the cell from desiccation as it is hygroscopic and drying.
- Attachment: capsule helps in attachment on diverse surface (of host) even on bacteriophage.
- Capsule protects bacteria from mechanical injury, temperature, drying.
- Capsule resists phagocytosis by WBCs hence it is anti-phagocytic.
- Capsule is a source of nutrition whenever nutrient supply gets very low in the cell.
- mutants use their capsules as a source of energy.
Flagella is a filamentous (hair-like) extension through the membrane or the wall of a cell. They vary from 15-20nm in diameter and help in motility of the bacteria (Figure 13).
These are unbranched, long, and mostly composed of the protein flagellin (known as H antigen). Motility plays important roles in survival and virulence of the bacteria.
Each flagellum consists of three main parts: filament, hook and basal body
Filament is a thin hair like structure that arises from hook and extends from the cell surface.
It is a wider region at the base of filament, embedded in the cell envelope, and acts as a flexible coupling between the filament and the basal body of the flagella.
It consists of central rod inserted into several rings attached to cell membrane and cell wall.
Based on arrangement of flagella they are divided into five types shown below:
- Monotrichous: single flagella attached on one end of the cell. Examples; Vibrio cholera, Pseudomonas
- Lophotrichous: bundle of flagella present in one end of the cell. Example: Pseudomanas
- Amphitrichous: single or cluster of flagella present at both the end of cell. Example; Aquaspirillium.
- Peritrichous: flagella present all over the surface of the cell. Example; E. coli, Salmonella, Klebsiella.
- Atrichous: flagella absent. Example, Shigella
Bacterial cytoplasm is composed of water, enzymes, nutrients, gases. It contains cell structures, such as ribosomes, chromosome, polypeptide (chaperones), cytoplasmic inclusions (volutin, polysaccharides, lipids etc.), and plasmids.
Some bacteria also produce spores and cysts.
Plasmids replicate independently of chromosome and appear to give bacteria a selective advantage to the bacteria.
Pili or Fimbriae are hair-like filaments (tiny hollow projections) that protrude from the cell membrane of the bacteria into the external environment.
They are usually present in gram-negative bacteria. A pilus is composed of subunits of the protein pilin. They play a significant role in adhesion and conjugation.
Some pili are useful in biofilm formation, phage transduction, DNA uptake, and bacterial motility.
Although bacteria are single-celled microscopic organisms, they reproduce in suitable conditions. Bacteria reproduce by vegetative, asexual, and sexual methods.
Vegetative reproduction includes binary fission, fragmentation, and budding. Sporulation (spore formation) is asexual form of reproduction in bacteria.
Sexual reproduction is by genetic recombination by three main processes: Conjugation, transduction, and transformation.
Binary fission is the commonest form of propagation for bacteria, where parent cell divides into two similar daughter cells.
During this process, an organism duplicates its genetic material (DNA) and then undergo a process of cytokinesis (division of cytoplasm) forming two daughter cells. Each daughter cell receives one copy of DNA.
The cell membrane starts growing inwards and by cytokinesis two daughter clone cells are formed — each containing separate DNA molecules.
In suitable conditions, bacteria can double in number through binary fission in 10-20 minutes.
Fragmentation is commonly known as splitting, where parent organism divide into several pieces and each piece develops into a complete new individual.
Budding is a form of vegetative reproduction that results from the outgrowth of a part of a cell leading to a separation of the original organism into two individuals.
The parent cell forms a bud at one end and forms a nucleus for the bud by the process of mitosis. Later, the bud separates from the parent cell and forms a different organism.
During unfavorable conditions, metabolically-dormant structure called spores are formed by the process of sporulation (Figure 14).
This process is divided into several stages. In Bacillus subtilis, the entire process of sporulation takes 8 hours to complete.
Spores can be produced within (endospores) or outside (exospores) the bacterial cells.
Bacteria producing endospore are Bacillus, Clostridium, Paenibacillus, Sporosarcina and bacteria producing exospore are Methylosinus.
Bacterial conjugation is a process by which a bacterial cell transfers genetic material to another bacterial cell (Figure 15). The bacterial DNA that is transferred by this process is called F-Plasmid (F is for plasmid).
A cell that has a copy of the F-plasmid is called F-positive, F-plus or F+ cell, and is a donor cell, while a cell that does not contain F-plasmid is called F-negative, F-minus or F– cell, and is considered a recipient cell.
Transformation is the process by which foreign DNA is introduced into a cell (Figure 16). Bacterial transformation plays a very important role in genetic engineering and molecular cloning.
After transformation, the host cells may express the acquired genetic information, which may serve as a source of genetic diversity and potentially provide benefits to them (e.g., antibiotic resistance).
The natural process of transformation was first reported by Frederick Griffith in 1928. The first protocol for artificial transformation of E. coli was published by Mandel and Higa in 1970.
This method became the basis for chemical transformation. Later it was discovered that an electrical field application to the cells also enhances the uptake of DNA, called an electroporation.
Bacteriophage is most commonly used virus for bacterial transduction (Figure 17). Random or generalized and specialized are the two kinds of transduction.
The table below shows the major points of difference between the two.
|Discovery||Generalized transduction was discovered in 1952 by Norton Zinder and Joshua Lederberg||Morse discovered specialized transduction in 1956|
|Phages||Mediated by lytic phages||Mediated by lysogenic phages|
|Process involved||Phage accidently packages a random piece of bacterial DNA just before the bacterial lysis||Phage DNA gets integrated in the bacterial DNA. During the unfavorable conditions, the viral DNA excise and sometimes carries a part of bacterial DNA too|
|DNA transferred||Phages can carry any part of the bacterial chromosome||Phages carry only restricted part of the bacterial chromosome|
Bacteria are not always “bad”. A human body encounters both good and bad bacteria every day.
Bacteria is called “good” as they help us digest our food and protects us from infections, whereas bad or pathogenic bacteria make us sick or even lead to mortality.
Examples of Various Bacteria
A few examples of good/harmless bacteria are Escherichia Coli, Lactobacillus Johnsonii, Bacteroides Thetaiotamicron, Staphylococcus epidermidis, Bifidobacterium Longum, Viridans Streptococci etc.
Neisseria gonorrhoeae, Mycobacteria, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Clostridium tetani, Clostridium botulimum, Bacillus anthracis, Bordetella pertussis, Borrelia burgdorferi are a few examples of harmful/bad bacteria.