ILOs. 10/10/2016 Maha Fathy 2

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2 ILOs 1- List different components of bacterial cell. 2-Describe structure of cell wall of Gram +ve and ve bacteria 3-Recognize role of cell wall and cytoplasmic membrane in survival and growth of bacterial cell. 4-Compare between mycoplasma, spheroplasts, protoplasts, and L-forms. 5- Define mesosomes and inclusion granules. 6-Recogenize differences between flagella and pili regarding structure and function. 7- Mention structure and function of bacterial ribosomes. 8-Correlate between different bacterial cell structures (capsule, fimbriae) and bacterial virulence. 9-Mention how, when, and why some bacterial genera produce endospores. 10/10/2016 Maha Fathy 2

3 Bacterial Structure Bacteria have characteristic cell structure (prokaryotic cells) Size: µm (µm= 1/1000 mm) Various shapes: spherical (cocci), rods (bacilli), spiral. 10/10/2016 Maha Fathy 3

4 Bacterial Shapes Cocci Bacilli Spirochetes Cocco Curved Bacilli Bacilli 10/10/2016 Maha Fathy 4

5 Bacterial Cell Structure Cell Wall Extra cellular structures Endospore Cytoplasm + Intra-cytoplasmic structures Cell (cytoplasmic) membrane 10/10/2016 Maha Fthy 5

6 Prokaryotic Cell Structure 10/10/2016 Maha Fathy 6

7 Cell Envelope Cell Wall Cytoplasmic Membrane Capsule & Slime Flagella Fimbria (Pili) 10/10/2016 Maha Fathy 7

8 Cell Wall Structure Give the bacteria its shape (rigid structure) Gram positive Gram negative 10/10/2016 Maha Fathy 8

9 Cell Wall Gram Positive Bacteria Thick peptidoglycan layer (repeating sugar subunits linked by peptide side chains) Teichoic acid (Polymers of glycerol or ribitol phosphate Major surface Ag) Gram Negative Bacteria Thin peptidoglycan layer Outer membrane formed of Lipopolysaccharides (Endotoxins) Peri-plasmic space 10/10/2016 Maha Fathy 9

10 Cell Wall of Gram positive Bacteria 40 sheets 10/10/2016 Maha Fathy 10

11 Peptidoglycan Structure (murein or mucopeptide) 10/10/2016 Maha Fthy 11

12 Cell Wall of Gram negative Bacteria 10/10/2016 Maha Fathy 12

13 Outer Membrane Structure 10/10/2016 Maha Fthy 13

14 Lipopolysaccharides of Gram negative bacteria 10/10/2016 Maha Fthy 14

15 Cell Wall Function Maintains the shape of the bacterial cell (cocci, bacilli etc). Supports the weak cytoplasmic membrane. Plays a role in cell division. It contains associated proteins: porins for selective permeability & adhesins for attachment Responsible for staining properties of the organism. 10/10/2016 Maha Fthy 15

16 Cell wall deficient bacteria Naturally occurring Mycoplasma Sterol containing plasma membrane Spontaneously (or artificially) induced) L-Forms Lab Induced Protoplast Spheroplast 10/10/2016 Maha Fathy 16

17 Cell wall deficient Bacteria Naturally occurring: Mycoplasma Smallest type of bacteria No defined cell shape Resistant to antibiotics which act on cell wall Resistant to lysozyme 10/10/2016 Maha Fathy 17

18 Lysozyme breaks this bond 10/10/2016 Maha Fthy 18

19 Lab induced cell wall deficient bacteria Protoplast Originate from Gram Positive bacteria Spheroplast Originate form Gram negative bacteria Induced by lysozyme which destroys the peptidoglycan layer Cell membrane Outer membrane 10/10/2016 Maha Fathy 19

20 Protoplast From Gram positive bacteria Induced by lysozyme All cell wall material has been removed Spherical structure surrounded by one membrane Spheroplast From Gram negative bacteria Induced by action of lysozyme on EDTA-treated Gram negative cell (or action of antibiotics) Still retaining part of cell wall (outer membrane) Spherical structure surrounded by two membranes 10/10/2016 Maha Fathy 20

21 How are these cell wall deficient bacteria kept without rupture??? By being maintained in an osmotically balanced medium (A solution that has the same concentration of ions and small molecules as cytoplasm) 10/10/2016 Maha Fathy 21

22 L-Forms Can occur Spontaneously (or induced) from some bacterial genera ( Bacillus, Clostridium, Haemophilus, Pseudomonas, Staphylococcus, and Vibrio). By action of antibiotics which can inhibit cell wall synthesis NB: First discovered in Lister Institute in London 10/10/2016 Maha Fathy 22

23 Medical importance of L-forms L-forms can remain viable inside the host They can replicate They are not killed by antibiotics acting on cell wall L-forms can revert to the normal form on removal of the inhibitor producing relapses of infection. 10/10/2016 Maha Fathy 23

24 Functions of Outer Membrane It protects the peptidoglycan from effects of lysozyme It impedes the ingress of many antibiotics It is associated with proteins: Porins: allow diffusion of some material Adhesins: attachment 10/10/2016 Maha Fathy 24

25 Cytoplasmic Membrane Composed of protein and phospholipids In contrast to eukaryotic cells: it is rich in protein with little phospholipids (no sterol except in mycoplasma) It has multifunctional nature Function: Active transport and selective permeability Involved in protein secretion (No Golgi apparatus nor endoplasmic reticulum in prokaryotic cell) Electron transport and oxidative phosphorylation (No mitochondria in prokaryotic cell) 10/10/2016 Maha Fathy 25

26 Mesosomes They are invaginations of the cytoplasmic membrane inside the cytoplasm. Two types of mesosomes are known as lateral and septal. Septal mesosomes are involved in cell division where bacterial DNA is attached. 10/10/2016 Maha Fathy 26

27 Role of septal mesosomes in cell division 10/10/2016 Maha Fathy 27

28 Capsule Found in some bacteria Mostly formed of high molecular weight polysaccharides (antigenic) or poly peptide Can not be seen in Gram stain preparations (special stain) An important virulence factor Capsular antigens are used for diagnosis & Typing 10/10/2016 Maha Fathy 28

29 The capsule as an important virulence Antiphagocytic Help in attachment of bacteria to mucous membranes factor Formed only in vivo 10/10/2016 Maha Fathy 29

30 Slime (Glycocalyx) Excreted by some bacteria as loosely attached polysaccharide material Plays an important role in attachment of bacteria to surfaces and formation of biofilms on biotic or abiotic surfaces It plays an important role in pathogenesis of many diseases (as cystic fibrosis and dental caries) Can lead to healthcare associated infection when attached to devices inserted in human host as IVDs or urinary catheters or prosthetics like artificial valves 10/10/2016 Maha Fathy 30

31 Flagella & Fimbria (Pili) 10/10/2016 Maha Fathy 31

32 Flagella Formed of proteins (Flagellin) Found in some bacteria Make bacteria motile (organelle of motility) Fimbria (Pili) Formed of proteins (Pilin) Found in some bacteria They are important virulence factors enabling bacteria to adhere to mammalian cells (colonization) Specialized pili are involved in gene transfer (sex pilus) in a process called conjugation 10/10/2016 Maha Fathy 32

33 Polar/Monotrichous Lophotrichous Amphitrichous Peritrichous 10/10/2016 Maha Fathy 33

34 Intra cytoplasmic structures 10/10/2016 Maha Fathy 34

35 Intracellular Structures Genetic material Storage granules or inclusion granules Ribosomes: protein synthesis Endospores 10/10/2016 Maha Fathy 35

36 Genetic material Bacterial chromosome -Single circular chromosome (double stranded DNA molecule) -Carries the genes essential for bacterial life Free in cytoplasm No nuclear membrane No nucleolus No histones No introns Extra chromosomal DNA: plasmid 10/10/2016 Maha Fathy 36

37 Plasmid Extra chromosomal double stranded circular DNA. Carry certain genetic information, e.g. antibiotic resistance, toxin production Can be transmitted from bacterial cell to another (gene transfer) Genetic information carried by plasmid are not essential for bacterial life 10/10/2016 Maha Fathy 37

38 Inclusion granules Round granules observed in cytoplasm of many bacteria. These are not permanent or essential structures. They are either stored energy or nutrient reserve concerned with cell metabolism, e.g. volutin granules (metachromatic granules of Corynebacterium diphtheriae). 10/10/2016 Maha Fathy 38

39 Ribosomes The site of protein synthesis. They have sedimentation coefficient of 70S being composed of 2 subunits: small 30S and large 50S subunits. Aggregates of multiple ribosomes on a m-rna are called polysomes Human cell ribosomes are 80S; formed of 2 subunits: 60S & 40S 10/10/2016 Maha Fathy 39

40 Endospores Metabolically dormant forms of some species (Bacillus & Clostridia) Spore formation occurs in response to unfavorable conditions, e.g. depletion of nutrition, accumulation of metabolites or unsuitable gaseous environment They are highly resistant to dryness, heat and chemicals as disinfectants They can live in soil for many years 10/10/2016 Maha Fathy 40

41 Spores are formed outside the body, Can not be stained by ordinary stains. Spore may be oval or round; terminal, central, or sub-terminal In germination each spore gives rise to a single vegetative cell which can multiply. 10/10/2016 Maha Fathy 41

42 Sporulation process Septum from invagination of cell membrane Fore spore Endospore maturation Death of mother cell and release of endospore 10/10/2016 Maha Fathy 42

43 Endospore structure The outer proteinaceous coat surrounding the spore provides much of the chemical and enzymatic resistance. Cortex: a very thick layer of specialized peptidoglycan A germ cell wall: a layer of peptidoglycan will become the cell wall of the bacterium after the endospore germinates Core: cell's DNA, ribosomes and large amounts of dipicolinic acid Additional outer glycoprotein layer called the exosporium. 10/10/2016 Maha Fathy 43

44 Resistance of Endospores Marked resistance of endospores is attributed to: Thick outer coats Impermeable cortex High content of calcium and dipicolinic acid Low water content Very low metabolic and enzymatic activity 10/10/2016 Maha Fathy 44

45 Gram positive Gram negative Peptidoglycan layer Thick Thin Teichoic acid Present Absent Outer membrane Absent Present Periplasmic space Absent Present LPS Absent Present Lipid and lipoprotein content Low (except mycobacteria) High Toxins Exotoxins Endotoxins & Exotoxins Disruption by lysozymes High Low Susceptibility to penicillins (in general) High Low Resistance to drying High Low 10/10/2016 Maha Fathy 45

46 Example Prokaryotic cell Bacterial cell Eukaryotic cell Fungal cell Human cell Cell wall peptidoglycan Present Absent Cell membrane Nuclear material No sterol (except mycoplasma) Single chromosome Extrachromosomal DNA No histones No introns Sterol Mitochondria Absent Present Rough endoplasmic reticulum Absent Present Ribosomes 70 S 80 S True nucleus Nuclear membrane Multiple chromosomes DNA associated with histones Introns Division 10/10/2016 Binary fission Maha Fathy Mitosis 46

47 Most welcome for any question 10/10/2016 Maha Fathy 47

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