Lecture 2: Bacterial Anatomy

Bacterial cells are characterized by complicated structure and are composed of different elements.

Structural elements of bacterial cell: Constant Cell wall Cytoplasmic membrane Cytoplasm Ribosomes Mesosomes Nucleoid Inconstant Capsule Flagella Spores Intracellular inclusions Pilli Plasmids

All structural elements of bacteria are sub classified into 2 groups: 1. Group of constant elements- elements of this group are always present in bacterial cell and may be detected in structure of bacteria belonging to different species and genera. Constant elements are the cell wall, cytoplasmic membrane, cytoplasm, ribosomes, mesosomes and nucleoid. 2. Group of inconstant elements includes- elements which may present or absent in bacterial cell. Some of them appear under unfavourable environment conditions such as dessication, starvation, changing of tempersature. Some structural

The cell wall takes part in cell division by forming an ingrowth from the cell wall. intracellular inclusions. flagella. Structures enveloping the bacteria. fimbria/pili and plasmids. . comprising the cytoplasma and the single circular chromosome of DNA. It is elastic. gelatinous layer known as capsule. The outer layer covering the bacteria cell consists of 2 components. 1. Inconstant elements are capsule.elements such as the flagella and fimbria are detected only in structure of certain genus and species of microorganism. The chemical composition for cell wall of Gram (+ve) and Gram (-ve) bacteria is different. Chemical Structure of the Cell Wall The cell wall is a tough and rigid structure surrounding the bacteria like a shell. The cell wall encloses the protoplasm. Cell wall of Gram (-ve) bacteria.the rigid cell wall and the cytoplasmic membrane. Some bacteria may possess additional structures such as the proctective. porous and freely permeable layer. spores.

The lipoprotein layer connects the outer membrane to the peptidoglycan layer. These specific proteins form porines and hydrophilic molecules are transported through these porines. The outer membrane is a phospholipids bilayer. In Gram (+ve) bacteria’s cell wall. The thickness of the peptidoglycan layer is 18-80 nm in Gram (+ve) bacteria while it is only 2 nm in Gram (-ve) bacteria. The polysaccharide represents a major surface antigen. connect bacteria with the environment through channels and pores. The toxicity is associated with the lipid portion. protection of cell from mechanical and osmotic action of the environment. Protoplasts are derived from Gram (+ve) bacteria. But sometimes the bacteria may lose cell wall and form derivatives: spheroplasts. Cell wall of Gram (+ve) bacteria. They are produced by growth of bacteria with penicillin. These acids are water soluble polymers. They are produced artificially by lysosome/ penicillin action. 2. Also. the O-antigen. The lipopolysaccharides layer is the endotoxin of Gram (-ve) bacteria. They are also unstable and osmotically fagile.The Gram(-ve) bacteria cell wall is a complex structure which containing 3 component layers outside the peptidoglycan layer. They lose their cell wall completely. the structure of the Gram (-ve) bacteria is characterized by the presence of periplasmic space. contains receptors for bacteriophages attachment (viruses of bacteria). This layer is attached to polysaccharides. Most Gram (+ve) bacteria cell wall contains a significant anount of teichoic and teichuronic acids. Hypertonic media are necessary for their maintenance. Functions of the cell wall: • • • • • • maintenance of the cell shape. The layer of lipopolysacchariedes is the third layer and it consist of a complex lipid called lipid A. it contains specific proteins. protoplasts and L-forms Spheroplasts are derived from Gram (-ve) bacteria. . It is a space between the inner and outer membranes and contains a number of important proteins and oligosaccharides playing an important role in osmoregulation of cells. They are osmotically fragile and maintain in hypertonic medium/culture. the peptidoglycan layer is much thicker than those in Gram (-ve) bacteria. determines the antigenic characteristics of the bacteria. The periplasmic space is absent and the peptidoglycan layer is closely associated with cytoplasmic membrane. These derivatives lose their cell wall partially. taking part in the regulation of growth and division of cells. The teichoic acids constitutes the major surface antigen of Gram (+ve) bacteria.

The main properties of L-forms: • • • • • • • • similarity of morphological changes. spheroplasts and protoplasts are able reverse back to their initial forms. takes part in the cell division. L-forms. formation of stable and unstable L-forms. Electron microscopy shows 3 layers forming the cytoplasmic membrane: the central layer is composed of protein molecules and on its both sides are lipid molecules. similarity of cultural changes. Methods for cell wall demonstration: • • • • plasmolysis antigen-antibody reaction differential staining ( Gram’s staining or acid-fast staining) electron microscopy techinique Cytoplasmic membrane It is a thin. ability to long persistence (able to circulate in the body for a long time and cause chronic infection). The cytoplasmic membrane is characterized by semi-permeability and selective transport. transformation of Gram (+ve) in Gram (-ve). They develop either spontaneously or artificially under the action of penicillin or other agents. decreasing of virulence. Functions of the cytoplasmic membrane: • • • • transport of chemicals through the cytoplasmic membrane (active and passive). changes in antigenic properties. takes part in bacterial oxydation. ability to return to initial form. elastic layer lying beneath the cell wall.L-forms are cell wall deificient forms of bacteria (the cell wall is destroyed by the action of antibiotics and lytic enzymes) and they are more stable than spheroplasts and protoplasts. Cytoplasmic membrane separates the cytoplasm components from the external environment. . They may multiply by binary fission while spheroplasts and protoplasts can not. the chemicals concentration. The plasma membrane is visible in ultra thin section under the electron microscope. separating it from the cell’s cytoplasm.

intracellular inclusions). They have sedimentation coefficient . few membranous bodies of mesosomes. Method for demonstration of mesosomes: • electron microscopy. The septal mesosome is attached to bacterial chromosomes (nucleoid) and involved in DNA segregation and in formation of cross over walls during cell division. They are distributed throughout the cytoplasm. The unit of sedimentation equal to 70s (consist of 2 units. There is only 1 septal mesosome in a bacterial cell. multilaminated.g. Several lateral mesosomes are detected in the structure of bacteria. Ribosomes Ribosomes are complicated structures 10-20 nm in size.Methods for demonstration of the cytoplasmic membrane: • • plasmolysis electron microscopy Mesosomes Mesosomes are derivatives of cytoplasmic membrane. membranous sac-like bodies. They are convoluted. Ribosomes are the sites of protein synthesis. Methods for demonstration of the cytoplasm: • • simple method of staining complicated method of staining . Mesosomes are principle centers of respiratory enzymes storage in bacteria. It is also the place of localization of the bacterial nucleoid. Method for ribosome demonstration: • electron microscopy.70s. Cytoplasm The cytoplasm is a colloidal watery solution containing organic and inorganic dissolved chemicals. They are formed by convoluted unvaginations of the plasma membrane into the cytoplasm. The biological role is that it is the medium for chemical reactions to take place in bacterial cell. There are 2 types of mesosomes: lateral and septal. vacuoles and inconstant elements (e. Lateral mesosomes are centers of respiratory enzymes storage.30s unit and a 50s unit). It lacks mitochondria and chloroplasts and contains large number of ribosomal granules.

Its structure resembles the nucleus.• • plasmolysis electron microscopy Nucleoid The bacterial nucleoid is a thin fiber of double stranded DNA tightly coiled inside the cytoplasm. Burvy-Gience staining) Pathogenic micro-organism representatives: Encapsulated bacteria (always with capsule layer): . (virulence is the measure of pathogenicity) It act as an antigent. It enhances the virulence of the bacteria. Methods for demonstration of the nucleoid: • • • electron microscopy special method of staining PCR. It is the most external layer. The solid component is complicated of polysaccharides or polypeptides. Capsule The capsule is an outer covering of thick material surrounding the bacterial cell wall. The nuclear DNA doesn’t contain basic proteins. bacteria still alive without inconstant elements. It consist of 98% water and 2% of solids.polymerase chain reaction . the double stranded DNA is haploid. It protects the bacteria from phagocytosis and penetration of lytic enzymes. Its function is the maintenance of genetic information/ characteristics. but there is no nuclear membrane and small nucleus (micronucleus). phagocytes and enzymes. Methods for demonstration of the capsule: • • electron microscopy special staining (negative method of staining. DNA-DNA hybridization Inconstant Structural Elements The inconstant structural elements are not as important as constant elements. Functions of the capsule: • • • It serves as a protective covering against antibacterial substances such as bacteriophages.

cytoplasmic appendages protruding through the cell wall. Flagella Flagella are filamentous. mucous membrane infection.single polar flagellum located at one end. Streptococcus pyogenus ( pathogen of pyogenus infection of different localization). Flagella are the organs of locomotion and have characteristics patterns of distribution in the bacterial cells. ( the fastest microorganisms). They are unbranched. long structures composed of the protein.• • • Klebsiella pneumoniae (pathogen of pneumonia) Klebsiella ozaenae (pathogen of ozena ) Klebsiella rhinoscleromatis (pathogen of rhinoscleroma). Capsule forming bacteria (form capsule only in human / animal body): • • • • • • Bacillus anthracis (pathogen of anthrax) Streptococcus pneumoniae ( pathogen of lobal pneumonia) Yersinia pestis ( pathogen of plague) Francisella tularensis (pathogen of tularemia) Staphylococcus aureus (pathogen of skin lesion. internal organ infection. According to the localization of flagella. . flagellated bacteria may be classified into 4 groups: • monotrichous. infection of the GIT).flagellin.

(e. the bacteria grow everywhere and while in the other tube. and we will see that in 1 tube. the basal membrane is localized in cytoplasmic membrane and the number of basal membrane is only one. staining by silver impregnation technique (Morosov’s stain). The main substructures are namely flagellum/filament. during starvation. amphitrichous. The structure of the Gram (+ve) and Gram (-ve) bacteria are different.numerous flagella surrounding the body of the bacterium.a tuft of flagella at one or both ends. and electron microscopy.flagella located on both sides (at both polar ends of bacteria). lophotrichous.g.) Bacterial spores Bacterial spores are highly resistant dormant (resting) forms of bacteria. Bacterial spores are formed under unfavourable environmental conditions. [Examination (x40)] (ii) study growth of microorganism in semi-solid medium. hook and basal membrane. the flagella has 2 basal membrane because its cell wall is thinner and has hook and filament. Method for flagella demonstration: • direct methods: Loffler’s staining. . the bacteria grow in the line of inoculation. then incubate. In Gram (-ve) bacteria. In Gram (+ve) bacteria.• • • peritrichous. bacteria inoculated by injection. ( 2 tubes. • indirect method: method of motility detection (i) non-fixed smear preparation: crush drop technique and hanging drop technique.

evaporation of water. maturation stage. 4. the diameter of the spore is smaller than the diameter of the vegetative part.B! Spores are not forms of replication.dessication. Spore may localize in different parts of the vegetative form: • • • centrally subterminal –between septal and end of the vegetative form. Stage of cell wall formation. change in temperature. N. In Clostridium. the diameter of the spore is bigger than the diameter of the vegetative part. The duration of sporulation process is 18-20 hours. Endospores are small spherical or oval bodies. forespore stage. Bacteria form endospores and never form exospore (e.g.at the end of the vegetative form. but NOT replication. The sporulation process occurs in 4 stages: 1. 2. only rod shaped bacteria are sporing. termial. So. In Bacillus. action of penetrative radiation and action of UV light. the function of spores is protection form unfavourable conditions. Only Bacillus and Clostridium are able to form bacterial spores. Methods of bacterial spore demonstration: . action of chemicals. 3.) The spores are formed within the bacterial cell are named endospores. preparatory stage. fungi) Thus.

Clostridium perfringens (pathogen of gas gangrene infection) Clostridium novyi (pathogen of gas gangrene infection) Clostridium septicum (pathogen of gas gangrene infection) Clostridium histolyticum (pathogen of gas gangrene infection) Fimbria/ Pilli Fimbria are thin. Auesky staining (modification of acid-fast staining) Spore forming bacteria: • • • • • • • Bacillus anthracis ( pathogen of anthrax). and the energy that it stores is used during starvation. Sex fimbria take part in conjugation and is responsible for transmission of genetic information from 1 cell to another. They are typical structures of Gram (-ve) bacteria. .pillin There are 2 types of fimbria: common fimbria and sex fimbria. They are the sources of stored energy and are only present in some species of bacteria. Intracellular inclusion Intracellular inclusions are localized in the cytoplasm. Fimbria are arranged peritrichously. The number of fimbria is more than the number of flagella. Common fimbria are responsible for attachment of the micro-organism to the substrate (host cells or tissue. Methods for demonstration of fimbria: • electron microscopy. These granules are found in large quantities when the bacteria are grown in abundance of nutrient. Each bacteria possess around 1-4 sex fimbria.• • Gram’s staining. short filamentous appendages extruding from the cytoplasmic membrane. polysaccharides and element sulphur. Fimbria are composed of the protein.vegetative form stain violet for Gram (+ve) bacteria and the spore is still colourless. Clostridium botulinum ( pathogen of botulism).) Each bacteria possesses 100-500 common fimbria. The braight representative of bacteria with intracellular inclusions is Corynebacterium diphtheriae where it has volutin granules. volutin. They are chemically composed of lipids. Clostridium tetani (pathogen of tetanus). Methods for demonstration of intracellular inclusions: • electron microscopy.

• Neisser’s method of staining. .

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