1) Bacteria have a simple cellular structure lacking membrane-bound organelles. Their cell envelope consists of an outer cell membrane and a peptidoglycan cell wall.
2) Gram-positive bacteria have a thick peptidoglycan cell wall exterior to the cell membrane, while gram-negative bacteria have an additional outer membrane and a thinner peptidoglycan layer between the inner and outer membranes.
3) Some bacteria form highly resistant endospores during periods of environmental stress, allowing them to survive harsh conditions. Endospores have a dehydrated core surrounded by protective layers and can remain dormant for long periods.
1) Bacteria have a simple cellular structure lacking membrane-bound organelles. Their cell envelope consists of an outer cell membrane and a peptidoglycan cell wall.
2) Gram-positive bacteria have a thick peptidoglycan cell wall exterior to the cell membrane, while gram-negative bacteria have an additional outer membrane and a thinner peptidoglycan layer between the inner and outer membranes.
3) Some bacteria form highly resistant endospores during periods of environmental stress, allowing them to survive harsh conditions. Endospores have a dehydrated core surrounded by protective layers and can remain dormant for long periods.
1) Bacteria have a simple cellular structure lacking membrane-bound organelles. Their cell envelope consists of an outer cell membrane and a peptidoglycan cell wall.
2) Gram-positive bacteria have a thick peptidoglycan cell wall exterior to the cell membrane, while gram-negative bacteria have an additional outer membrane and a thinner peptidoglycan layer between the inner and outer membranes.
3) Some bacteria form highly resistant endospores during periods of environmental stress, allowing them to survive harsh conditions. Endospores have a dehydrated core surrounded by protective layers and can remain dormant for long periods.
GROWTH, AND METABOLISM • Phospholipid w/c form 2 parallel surfaces
(phospholipid bilayer) • Polar phosphate groups - outside 2 Basic Types of Microorganisms • Nonpolar lipid chains are – inside • Acts as permeability barrier, restricting Prokaryotic (Before – nucleus) molecules that enter and leave the cell Eukaryotic (True – nucleus) Peptidoglycan
• Determines the shape of the cell
Prokaryotes • Composed of a cross-linked polymeric mesh ➢ include the bacteria and cyanobacteria • Glycan – linear polymer of alternating which were formerly classified as blue- monosaccharide subunits (backbone) green algae. • Peptido – short string of AA forming a ➢ They possess a simple make-up that does network w/ high tensile strength not contain sub-cellular organelles. The typical size of a prokaryote is about 1um diameter. ➢ Archaebacteria and Eubacteria (MONERA) Difference b/w Gram (+) & Gram (-) Cell Walls:
➢ contain membrane-bound organelles walls that are exterior to the cytoplasmic such as mitochondria, lysosomes, (plasma) membrane endoplasmic reticulum and golgi bodies. • The peptidoglycan in most gram (+) species is covalently linked to teichoic acid, which is essentially a polymer of substituted glycerol units linked by 2 Kinds of Cells phosphodiester bonds Eukaryotic – cells with a well-defined nucleus • Teichoic acids are major cell surface antigens & are integrated into the Prokaryotic – cells without a nucleus peptidoglycan layers but not tethered to the All bacteria are prokaryotes cytoplasmic membrane • Lipoteichoic acids are lipid modified and Unorganized DNA (single double-stranded integrated by this moiety into the outer molecule) leaflet of the cytoplasmic membrane Gram (-) Bacteria Cell Wall Bacterial Structure • More complex cell wall structure composed of 2 membranes (an outer membrane and Cell Envelope a periplasmic membrane/space) (inner • Term applied to all material external to and membrane = cytoplasmic / plasma enclosing the cytoplasm membrane) • Layers: Cell Wall, Plasma / Cell / • The 2 membranes are separated by Cytoplasmic Membrane, * Capsule (some / periplasmic space, w/c contains the not all) peptidoglycan layer • It consists of several chemically and • The periplasmic space also contains functionally distinct layers, the most degradative enzymes and transport prominent of which are the cell wall and proteins the cytoplasmic membrane • The peptidoglycan layer of gram (-) cells is • Also includes the capsule or glycocalyx, if thin, and cells are consequently more present susceptible to physical damage • The outer membrane is distinguished by • These result in the formation of a dormant the presence of embedded cell called an endospore inside the original lipopolysaccharide (LPS) that is the cell major constituent of the outer leaflet of the • Endospores can be released from the outer membrane original cell as free spores • The polysaccharide portion of LPS (O- • Spores are the most resistant life forms polysaccharide) is antigenic and can, known therefore, be used to identify different • They are remarkably resistant to heat strains and species (they survive boiling), desiccation, • The lipid portion (lipid A) is imbedded in ultraviolet light, and bactericidal chemical the membrane and is toxic to humans and agents animals • In fact, sterilization procedures are • It is called endotoxin assessed by their ability to inactivate Spores External Capsule and Glycocalyx Sporulation • sticky, viscous, slime material that forms an extracellular coating around the cell – • Repackaging a copy of bacterial DNA into glycocalyx a new form that • allows cells to adhere to surfaces, protect - contains very little water bacteria from antibodies & phagocytosis - no metabolic activity • Capsules can also protect bacteria against - does not divide desiccation, or drying, which facilitates - has a restructured, highly impermeable, transmission multilayered envelope • Begins with the invagination of the parent Appendages cell membrane, producing a double Flagella membrane that encapsulates and isolates a copy of the bacterial DNA in what will • Long, semirigid, helical, hollow tubular become the core of the spore structures composed of several thousand • Mature spore retains the complete molecules of the protein flagellin machinery for protein synthesis, and new • Enable bacteria to move in a directed spore-specific enzymes are synthesized in fashion the core of the spore • Anchored in the cell membranes by a basal • The core also has high levels of a unique body, which is a complex molecular compound called calcium dipicolinate, machine that rotates the flagellum like the which is thought to be important for screw propeller of a ship protection of the spore DNA from • Cells may have one or many flagella environmental damage • Highly antigenic • Many enzymes of the original vegetative (non-dividing) cells are degraded Pili (Fimbriae) • When the endospore is completed, the • Shorter and thinner than flagella and parent cell lyses, releasing the spore function as attachment structures that Medical Significance of Sporulation promote specific cell-to-cell contact • The attachment can be between the • Some of the most notorious pathogens are bacterial cell and the host eukaryotic cell or spore-formers, including: between one bacterial cell and another - B. anthracis - Bacillus cereus (gastroenteritis) Spores and Sporulation - Clostridium tetani • To enhance survival during periods of - Clostridium botulinum environmental hostility (such as - Clostridium perfringens (gas gangrene) nutritional deprivation), some - Clostridium difficile gram-positive rods undergo profound • Spores of these organisms can remain structural and metabolic changes viable for many years and are generally not killed by boiling, but they can be killed by autoclaving (temps > 120oC at elevated • Because bacteria reproduce by binary pressure) fission (1 becomes 2, 2 become 4, 4 • In the absence of an autoclave, spores can become 8, etc.), the number of cells be largely eliminated by a primary boiling to increases exponentially with time (the activate germination and, after a short exponential, or log, phase of growth) period of vegetative growth, a second • Depending on the species, the minimum boiling doubling time can be as short as 10 mins or as long as several days • Ex: for a rapidly growing species such as E. Bacterial Growth and Metabolism coli in a nutritionally complete medium, a single cell can give rise to some 10 million ➢ All cells must accomplish certain cells in just 8 hours metabolic tasks to grow and divide • Eventually, growth slows and ceases ➢ All cells, whether bacterial or human, entirely (stationary phase) as nutrients accomplish these metabolic tasks by are depleted, and toxic waste products similar pathways accumulate ➢ There are, however, some important • Most cells in a stationary phase are not differences that set bacteria apart dead, however metabolically from eukaryotic cells • If they are diluted into fresh growth Characteristics of Bacterial Growth medium, exponential growth will resume after a lag phase • If bacterial cells are suspended in a liquid nutrient medium, the increase in cell Energy Production number or mass can be measured in • A distinctive feature of bacterial several ways metabolism is the variety of mechanisms • Techniques include: used to generate energy from carbon - microscopically counting the cells in a sources given volume using a ruled slide • Depending on the biochemical mechanism - counting the number of appropriately used, bacterial metabolism can be diluted cells that are able to form colonies categorized into 3 types: following transfer to a solid nutrient (agar) - aerobic respiration surface - anaerobic respiration - quantitating the turbidity—which is - fermentation proportional to the cell mass—of a culture in liquid medium Aerobic Respiration
Stages of Bacterial Growth Cycle • Metabolic process in which molecular
oxygen serves as the terminal electron acceptor of the electron transport chain • In this process, oxygen is reduced to water • Respiration is the energy-generating mode used by all aerobic bacteria Anaerobic Respiration
• Metabolic process in which inorganic
compounds other than molecular oxygen serve as the terminal electron acceptors • Depending on the species, acceptors can 1. Lag Phase be molecules such as nitrate or sulfate • Anaerobic respiration can be used as an 2. Log (exponential) Phase alternative to aerobic respiration in some 3. Stationary Phase species (facultative organisms), but is obligatory in other species (some obligate 4. Death (decline) Phase anaerobes) Fermentation • S. aureus - most virulent specie secrete coagulase, an enzyme that causes • Anaerobic process utilized by some citrated plasma to clot bacterial species • S. aureus 🡪 COAGULASE (+) • It is the metabolic process by which an organic metabolic intermediate derived Coagulase Test from a “fermentable” substrate serves as 🡪 Used to differentiate S. aureus from coagulase the final electron acceptor (-) species (S. epidermidis, S. saprophyticus) 🡪 2 drops saline 🡪 slide (T) and slide (C) MODULE 4: STAPHYLOCOCCI 🡪 Emulsified with the organism using a wire loop slide (T)
Gram Staining 🡪 A drop of plasma anticoagulated w/ EDTA
(EthyleneDiamineTetraacetic Acid) is placed on • Staining method using crystal violet the slide (T) & (C) (primary stain) & a counterstain – safranin 🡪 Macroscopic clumping in the plasma within 10 or fuchsine secs in slide (T) w/ no clumping in the slide (C) 🡪 • To distinguish & classify bacteria into Gram Coagulase (+) (+) or Gram (-) • Based on its peptidoglycan cell wall 🡪 No clumping in both slides 🡪 Coagulase (-) • Developed by Hans Christian Gram (1884) * followed by a tube test ➢ Gram (+) bacteria stains (DARK) VIOLET • Other species that occasionally cause (retains the Crystal Violet) disease and lack coagulase are often ➢ Gram (-) bacteria stains PINK or RED referred to as coagulase-negative (retains the counterstain Safranin or staphylococci Fuchsine) • Resistant to heat and drying; can persist ➢ Gram-variable / Gram-indeterminate 🡪 for long periods on fomites (inanimate unclear or no stain objects), which can then serve as sources - Ex: Typhus bacillus of infection Staphylococci General Features • Frequent hand-washing before and after contact with food or potentially infected • Stains dark violet 🡪 Gram (+) individuals decreases the transmission • Round and tend to occur in bunches like grapes Staphylococcus Aureus • Because growth of staphylococci requires • Generally, significant host compromise supplementation with various amino acids is required like: and other growth factors, they are routinely - break in the skin or insertion of a foreign cultured on enriched media containing body (ex: wounds, surgical infections) nutrient broth and/or blood - obstructed hair follicle (folliculitis) • Facultatively anaerobic organisms - compromised immune system • Produce catalase 🡪 Catalase (+) Disease may be: Catalase Test 1) largely or wholly the result of actual (+) Bubbles 🡪 Catalase (+) 🡪 Staphylococci invasive infection, overcoming host defense mechanisms, and the production (-) Bubbles 🡪 Catalase (-) 🡪 Streptococci of extracellular substances which facilitate 🡪 A small drop of HYDROGEN PEROXIDE is invasion placed on a microscope slide 2) a result of toxins in the absence of invasive infection (“pure” toxinoses) 🡪 Applicator stick is touched to a bacterial colony 3) a combination of invasive infection and and the tip is smeared onto the hydrogen peroxide Intoxication drop 🡪 Fibroblasts and their products may form a wall around the inflamed area, which contains bacteria Epidemiology and leukocytes • Frequently carried by healthy individuals on 🡪 This creates a characteristic pus-filled boil or the skin and mucous membranes abscess • Carriers serve as a source of infection to themselves and others (ex: direct contact, 🡪 Serious consequences occur when the bacteria by contamination of fomites (objects such invade the bloodstream 🡪 septicemia (presence as a doorknob, which in turn can be a and persistence of pathogenic microorganisms source of infection) or contamination of or their toxins in the blood) 🡪 rapidly fatal food, which can then result in food 🡪 Bacteremia (presence of viable bacteria poisoning circulating in the bloodstream) may result in Pathogenesis - seeding internal abscesses Cell wall virulence factors: - skin lesions - Capsule: thin (microcapsule) & resistant to - infections in the lung, kidney, heart, skeletal phagocytosis muscle, or meninges - Protein A: major component of cell wall 1. Localized skin infections - Fibronectin-binding protein (FnBP) 🡪 adhesin 🡪 - small, superficial abscesses involving hair promote binding to mucosal cells & tissue follicles (folliculitis) or sweat or sebaceous glands - Clumping factor: Coagulase - Ex: sty (external hordeolum) – eyelash 🡪 Cytolytic exotoxins: attack RBC (hemolysin) - Furuncle (boil) – subcutaneous abscess 🡪 Panton-Valentine leukocidin: pore-forming (neck, face, axilla, buttocks) toxin making the strain more virulent (community- - Carbuncle – larger, deeper skin infections acquired methicillin-resistant S. aureus – MRSA) - Impetigo – localized, superficial, spreading 🡪 Superantigen exotoxins crusty skin lesion in children - Enterotoxins: cause food poisoning 2. Deep, localized infections - Toxic shock syndrome toxin (TSST-1) - metastatic from superficial infections - Exfoliatin toxin (ET): scalded skin syndrome in - most common cause of acute and chronic children bone marrow infection Clinical Significance - most common cause of acute infection of 🡪 S. aureus causes disease by infecting tissues, joint space in children (septic joint) – emergency typically creating abscesses and/or by producing because pus can rapidly cause irreparable toxins cartilage damage
🡪 A common entry point into the body is a break 3. Acute endocarditis
in the skin, which may be a minute needlestick or - generally associated with IV drug abuse by a surgical wound injection of contaminated preparations or needles 🡪 Another portal of entry is the respiratory tract - also colonizes skin around the injection site, (Ex: staphylococcal pneumonia) and if the skin is not sterilized before injection, 🡪 Localized host response to staphylococcal bacteria can be introduced into soft tissues and the infection: bloodstream, even when a sterilized needle is used - inflammation (swelling) 4. Septicemia - accumulation of pus (pyogenic) - generalized infection with sepsis or bacteremia - necrosis of tissue that may be associated with a known focus (for example, a septic joint) or not (an occult focus) 5. Pneumonia Immunity 6. Nosocomial infections 🡪 Do not elicit strong or long-lasting immunity, as demonstrated by the continuing susceptibility of - hospital-associated infections, often of individuals to S. aureus infections throughout life wounds (surgical, decubital) or bacteremia associated with catheters Treatment 7. Toxinoses 🡪 Require aggressive treatment, including incision and drainage (I&D) of localized lesions, as well as a. Toxic shock syndrome: systemic antibiotics - high fever - Oxacillins - rash ('sunburn', w/ diffuse erythema followed - Cephalosporins by desquamation) 🡪 Vancomycin for methicillin-resistant S. aureus - vomiting, diarrhea 🡪 hypotension Coagulase-negative Staphylococci - multiorgan involvement (GI, renal, and/or hepatic damage) 🡪 Of 12 species recovered as normal commensals of human skin and anterior nares, the most b. Staphylococcal gastroenteritis: abundant and important is S. epidermidis - ingestion of food contaminated with 🡪 The second most important coagulase-negative enterotoxin staphylococcus is S. saprophyticus - protein rich (egg salad or cream pastry) or 🡪 Important agents of hospital-acquired infections salty, like ham, and improperly refrigerated associated with the use of implanted prosthetic - nausea, vomiting, and diarrhea, are acute devices and catheters following a short incubation period (< 6 hours) Staphylococcus Epidermidis because toxin in the food has already been formed before food is ingested • Present in large numbers as part of the normal flora of the skin c. Scalded skin syndrome: • As such, it is frequently recovered from - superficial bullae resulting from the action of blood cultures, generally as a contaminant an exfoliative toxin that attacks the intercellular from skin adhesive of the stratum granulosum, causing • Despite its low virulence, it is a common marked epithelial desquamation cause of infection of implants such as heart valves and catheters - the bullae may be infected or may result from • Acquired drug resistance by S. epidermidis toxin produced by organisms infecting a different is even more frequent than by S. aureus site • Produces an extracellular polysaccharide Lab Identification material called polysaccharide intercellular adhesin (“slime”), that facilitates adherence 🡪 Stain strongly gram (+), and are frequently seen to bioprosthetic material surfaces, such as in grapelike clusters intravenous catheters, and acts as a barrier 🡪 Relies largely on microscopic and colony to antimicrobial agents morphology and catalase positive Staphylococcus Saprophyticus 🡪 Distinguished from the coagulase-negative staphylococci primarily by coagulase positive • Frequent cause of cystitis in women, probably related to its occurrence as part of 🡪 Tend to be yellow (“aureus,”- golden) and normal vaginal flora hemolytic, rather than gray and nonhemolytic like • It tends to be sensitive to most antibiotics, the coagulase-negative staphylococci even penicillin G • Distinguished from S. epidermidis and 🡪 S. aureus is also distinguished from most most other coagulase-negative coagulase-negative staphylococci by being staphylococci by its natural resistance to mannitol-positive novobiocin MODULE 5: STREPTOCOCCI Serologic (Lancefield) Groupings 🡪 Many species of streptococci have a polysaccharide in their cell walls known as C- Streptococci General Features substance, which is antigenic and easily extractable with dilute acid • Gram (+) • Non-motile 🡪 Classifies primarily β-hemolytic streptococci • Catalase (-) into groups A through U on the basis of their C- • Ovoid to spherical substance • Pairs or chains 🡪 The clinically most important groups of β- • Aerotolerant / Facultative anaerobes hemolytic streptococci are: • Blood enriched medium - Groups A and B 2 Ways of Classification Group A β-Hemolytic Streptococci (GABHS) 1.) Hemolytic properties on blood agar 🡪 S. pyogenes (Streptococcus Pyogenes) 2.) Serologic (Lancefield) groupings • most clinically important member of this Hemolytic Properties on Blood Agar group of gram (+) cocci ➢ α-Hemolytic: chemical change in the • one of the most frequently encountered hemoglobin in blood agar, resulting in a bacterial pathogens of humans worldwide green pigment that forms a ring around • can invade apparently intact skin or the colony mucous membranes, causing some of the most rapidly progressive infections known • cause rheumatic fever and acute glomerulonephritis • Nasopharyngeal carriage is common especially in colder months and particularly among children • does not survive well in the environment (unlike staph) • instead, its habitat is infected patients and also normal human carriers in whom the organism resides on skin and mucous membranes ➢ β-Hemolytic: gross lysis of RBC, resulting • usually spread person to person by skin in a clear ring around the colony contact and via the respiratory tract ➢ γ-Hemolytic: is a term applied to • usually form long chains when recovered streptococci that cause no color change from liquid culture but may appear as or lysis of RBC individual cocci, pairs, or clusters of cells in Gram stains of samples from infected tissue Capsule: 🡪 Hyaluronic acid (Hyaluronan), identical to that found in human connective tissue, forms the outermost layer of the cell 🡪 Not recognized as foreign by the body and, therefore, is nonimmunogenic 🡪 Antiphagocytic Cell Wall: - treated with a topical agent such as mupirocin, or systemically with penicillin or a first-generation 🡪 contains a number of clinically important cephalosporin (cephalexin) components: 🡪 Erysipelas ➢ M Protein (evasive protein) - not infectious if absent - affecting all age groups - highly variable, w/ 80 different antigenic - fiery red, advancing erythema, especially on types the face or lower limbs - antiphagocytic ➢ Protein F (fibronectin-binding protein) 🡪 Puerperal sepsis - mediates adhesion / attachment in the pharyngeal epithelium - initiated during, or following soon after, delivery caused by exogenous transmission (nasal droplets Extracellular Products: from an infected carrier or from contaminated instruments) or endogenously, from the mother’s 🡪 secretes a wide range of exotoxins that often vaginal flora vary from one strain to another 🡪 Streptococcal toxic shock syndrome Epidemiology: - mediated by production of pyrogenic exotoxins 🡪 the only known reservoir is the skin and mucous membranes of the human host - initially flulike symptoms, followed shortly by necrotizing soft tissue infection, shock, acute 🡪 respiratory droplets or skin contact spreads from respiratory distress syndrome, and renal failure person to person, especially in crowded environments (classrooms & play areas) - Tx: penicillin G + clindamycin Clinical Significance 🡪 Post-streptococcal sequelae 🡪 Major cause of cellulitis - Acute rheumatic fever after 2-3 weeks of pharyngitis characterized by fever, rash, carditis, & 🡪 Acute tonsillopharyngitis (ATP) / arthritis pharyngotonsillitis - Acute glomerulonephritis after a week of - most common type of infection impetigo or pharyngitis causing renal failure - “strep throat” is associated with severe, Lab Identification purulent inflammation of the posterior oropharynx and tonsillar areas 🡪 Rapid latex antigen kits
• (+) test: latex particles clump together
• (-) test, they stay separate, giving the suspension a milky appearance Treatment 🡪 Penicillin 🡪 Clarithromycin or Azithromycin 🡪 Plus Clindamycin to inhibit protein (toxin) synthesis released from rapidly dying bacteria Group B β-Hemolytic Streptococci (GBBHS) 🡪 Impetigo 🡪 S. Agalactiae (Streptococcus Agalactiae) - classic cause • gram (+), catalase (-) - begins on any exposed surface (legs) • found in vaginocervical tract of female - extensive lesions on face & limbs carriers, and the urethral mucous membranes of male carriers as well as in the gastrointestinal (GI) tract • can be transmitted sexually among adults • capsular swelling observed when reacted and from an infected mother to her infant at with type-specific antisera (Quelling birth reaction) • leading cause of meningitis and septicemia • most common cause of community- in neonates, with a high mortality rate acquired pneumonia and adult bacterial • occasional cause of infections in meningitis and is an important cause of postpartum women (endometritis) and otitis media, sinusitis and mastoiditis individuals with impaired immune systems, • Fastidious (complex nutritional in whom the organism may cause requirements) and routinely cultured on septicemia or pneumonia blood agar • Latex agglutination tests can also • Releases an α hemolysin that damages demonstrate the presence of group B red cell membranes, causing colonies to antigen in these samples be α hemolytic • Group B streptococci are β hemolytic, with larger colonies and less hemolysis than group A • Most remain sensitive to penicillin G and ampicillin, which are still the antibiotics of choice 🡪 S. Pneumoniae (Streptococcus Pneumoniae) (Pneumococcus)
• gram-positive, nonmotile, encapsulated
cocci • lancet shaped, and their tendency to occur in pairs accounts for their earlier designation as Diplococcus pneumoniae