Mammary Immunology 2
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| CMT | Measures nuclear DNA, more cells = more DNA = stickier = more clumping
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| SCC Beneficial | Migrate rapidly and in sufficient numbers during early stages of infection, function properly and eliminate bacteria effectively, return to low levels quickly
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| SCC Detrimental | Delayed migration, unable to control rapid growth of bacteria, elevated numbers persist for a long period of time
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| N-acetyl-B-D Glucosaminidase (NAGase) | Lysosomal enzyme increased during mastitis, comes from immune cells
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| Mastitis Control Program | Udder health goals, clean/dry environment, proper milking procedure, milking equipment, record keeping, clinical mastitis management, dry cow management, biosecurity, monitor udder health, review
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| Inhibitors of Cell Wall Synthesis | Beta-lactams (penicillins and cephalosporins), polypeptides (bacitracin), and vancomycin - inhibit when bacteria is actively growing
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| Inhibitors of Protein Synthesis | Macrolides (erythromycin), lincosamides (clindamycin), and chloramphenicol bind 50s subunit, animonglycosides (genatmycin, amikacin) and tetracyclines bind 30s subunit - binds ribosomes to prevent function
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| Prevent Transcription or DNA Integrity | Quinolones (norfloxacin, ciprofloxacin, enroflocacin) inhibit DNA gyrase or topoisomerase, rifamycins (rifampin) inhibit mRNA formation and transcription
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| Antibiotics | Time and concentration dependent, antimicrobial resistance dependent
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| Gene Transfer | Transduction: through phage or virus, conjugation: through pili, competance: release of DNA into the environment and accepted through phagocytosis
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| Resistance | Microbe produces enzyme that destroys antibiotic, microbial enzyme added to antibiotic structure, changes in cell wall permeability, changes in target protein, transport or antibiotic out of cell
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| Contagious Pathogens | Streptococcus agalactiae (eradicated bc obligate of mammary gland), Staphylococcus aureus, Corynebacterium bovis (monitoring tool) - source usually infected udders, transmission at milking, usually chronic subclinical
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| Environmental Pathogens | Source is cows environment, transmission between milkings, higher incidence in herds that control contagious pathogens, infections during dry period and at calving, less responsive to mastitis control practices
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| Opportunistic Pathogens | Source is healthy teat skin + milkers hands, transmission due to bacterial load on teat end, most common bacteria in herd, no effect on milk production, eliminated with teat dip and dry cow therapy, responsive to antibiotics, protection from others
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| Uncommon Mastitis Pathogens | Psuedomonas aeruginosa from contaminated water/teat dip, Nocardia species from soil and bad infusions, Actinomyces pyogenes from soil, Mycoplasma species contagious, preceded by respiratory disease
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| Staphylococcal Species | Catalase positive, produce catalase enzyme, degrade hydrogen peroxide to water and O2, gram + cocci, haemolysis (a green, b clear zone, y none), thick cell wall, SA coagulase positive
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| Pathogenicity | Ability to infect a host
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| Pathogenic | Disease causing
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| Virulence | Ability to cause disease in the infected host, bacterial properties that enhance disease-producing capabilities, avirulent = cant cause disease
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| S. aureus | Gram (+) cocci, colonize skin, nasal cavity, and mucosal membranes, thrive on unhealthy skin, coagulase positive, b haemolysis - pathogen wins 70-80% of the time, cause clots = shed in cyclic manner
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| Abscess Formation by SA | Host attempts for confine bacteria, wall off bacteria with scar tissue = poor antibiotic cure rates, SA can break through accesses
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| Virulence Factors | Any characteristic the bacteria produces that can increase the chance for infecting a host, causing disease, prolonging disease, or altering the severity of a disease
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| Surface Factors | Cell envelope teichoic/lipoteichoic acid, peptidoglycan, protein A, pseudocapsules, slime, adhesins,
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| Teichoic/Lipoteichoic Acid | AFFECTS ERYTHROCYTES, activates complement, hypersensitivity, adhesion to host cells, biofilm formation, covalently bound to peptidoglycan, specific antibodies - antigenic determinant
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| Peptidoglycan | Affected by growth conditions, pyrogenic effects (TNF, IL 1/6), aggregation and lysis of blood platelets, INHIBITS LEUKOCYTE MIGRATION, purulent lesion, specific antibodies, biosynthetic function, active transport
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| Protein A | Cell wall component, 90-98% SA, inhibits opsonization through Fc binding, blood coagulation, platelet damage, 1 protein A binds two Ig,, resists milk flushing
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| Pseudocapsules | Polysaccharide surface, frequently observed on fresh isolates and those grown in milk, acts as a PHYSICAL BARRIER for C3 and antigen masking, type 5 and 8 in 70-80% of strains, evade complement
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| Slime | Diffuse capsule, heterogeneous in composition, saccharides, neutral sugars and amino acids, poor antigen - no specific antibodies against it, anti-phagocytic
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| Adhesins | Surface protein: affinity for fibronectin, collagen, laminin, fibrinogen, resists flushing effect of milking
Cell wall anchored proteins: collagen and fibrinogen binding protein, clumping factor A and B, fibronectin binding proteins
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| Secreted Factors | Coagulase, enterotoxins, superantigen, haemolysins, A/B/Y toxins, hyaluronidase, lipase, catalase, nuclease, fatty acid modifying enzyme, Panton-Valentine leukocidin, staphylokinase
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| Coagulase | Protein that binds prothrombin and converts fibrinogen to fibrin, heterogeneous, encourages walling off of SA
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| Enterotoxins | Associated with foodborne illness, diarrhea and vomiting, 8 types, TSST-1, produced during infection by 4-83% of isolates, act as mitogen/superantigen causing nonspecific expansion of T cells
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| Superantigen | Overproduction of IL2, causes clonal expansion of all T cells nonspecifically
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| Haemolysins | A/B/Y/D four types, ability to lyse red blood cells, significance as virulence factors vary, CNS cannot perform haemolysis
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| Hyaluronidase | Break down tissue structure
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| A Toxin | Binds plasma membrane of epithelial cells, pore formation, osmotic lysis, teat cells more resistant, secretory cells more sensitive, septic shock
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| B Toxin | Makes cell more susceptible to A toxin, cytotoxic to secretory epithelial cells
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| Y Toxin | Binds and damages cell membranes, activates various cells
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| Panton-Valentine Leukocidin | 2 components acting in synergy, kills leukocytes by forming pore, produced by large number of isolates from acute dermatitis, less hemolytic than A toxin, microbes form of MAC, PVL + infections more severe
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| Staphylokinase | plasminogen activator protein, role in fibrinogen degradation, most mastitis strains do not produce this factor, degradation of clot - cyclical shedding, usually only in clinical cases
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| Gangrenous Mastitis | SA is main bacterial agent, rapid multiplication and toxin release, blood vessel constriction, massive blood clots, restricts blood supply to affected area
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| SA Diseases | Pneumonia, sepicemia, osteomyelitis/septic arthritis, toxic shock syndrome, wound infections, scalded skin syndrome
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| Colonization | Adhesins, binding proteins: fibronectin, collagen, laminin, fibrinogen, surface factors: techoic acid, protein A, peptidoglycan
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| Host Response | Lysozyme, lactoferrin, lactoperoxidase, complement, phagocytosis, antibody
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| Invasins | Staphylokinase, hyaluronidase, extracellular enzymes: proteases, lipases, nucleases, collagenase, elastase
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| Resistance to Phagocytosis | Coagulase, protein A, leukocidin, hemolysins, carotenoids, superoxide dismutase, catalase, antigenic variation, growth at low pH
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| Resistance to Immune Response | Coagulase, antigenic variation
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| Host Response | Cell activation (TCR MHCII binding), cytokine production/adhesion molecule, apoptosis
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| Toxigenesis | TSST toxin, enterotoxins A-G, cytotoxic toxins (hemolysins and leukocidin)
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| Host Response | Disease
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| MRSA | Small red bumps to abscesses, rapid blood test, SR assay using positive blood culture, PCR to test for SA and MR,
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| CNS Infections | Bacteremia, endocarditis, UTI, surgical site infections, endophthalmitis, colonization of indwelling plastic devices, infections in preterm infants
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| CNS Virulence | Capsular polysaccharide, biofilm formation, adhesins, protection from chemotaxis, phagocytosis, resistance to host microbial peptides, lipoteichoic acids and peptidoglycan, antibiotic resistance genes
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| Inactivated Vaccine | Virulent micro-organism killed with chemicals or heat - Polio, pertussis, influenza, J5
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| Live, Attenuated | Live miro-organism with disabled virulent properties but intact ability to simulate immunity - influenza and lyme
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| Toxoid | Inactivated toxic compounds from microoganisms - tetnus
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| Heterologous Vaccine | Microorganism that shares cross-reacting antigens with the microorganism that causes the disease - cowpox, smallpox, BCG vaccine for tuberculosis
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| Subunit | Fragment of an organism that can create an immune response - HepB
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| Goals of SA Vaccine | Eliminate chronic infections, reduce clinical mastitis, prevent new infections
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| Staph-Vax-Conjugate | Capsular polysaccharide 5+8 + pseudomonas exotoxin A, phase III human trials showed reduction in bacteremia but short lasting
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| A-Hemolysin | Protective against pneumonia in mouse model, prevents lung epithelial cell lysis, protected damage to lung which helped control bacterial growth and focused the immune response
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| PVL | Tested in same context as A hemolysin - not protective
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| RIP AGR Inhibitor | No toxin production, decreased biofilm formation, target protein within system can alter how SA produces and secretes toxins
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| Passive Immunotherapies | Hyperimmune IgG preparation, anti-lipoteichoic acid monoclonal antibody for initial use in preventing staph infections in neonates
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| Lysostaphin | Enzyme that selectively cuts through the cell wall of SA, cleaves polyglycine cross-links in the peptidoglycan layer of the cell wall of Staph species - success in transgenic species but expensive
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| Accessory Gene Regulator | Regulates the production of Staph toxins at late stages of growth, decreases production of surface adhesion proteins and increases secreted proteins
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| Heteropolymer Antibodies | Monocolonal antibody specific to a red blood cell receptor that is chemically linked to a second antibody that binds a particular pathogen
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| Mastitis Vaccines | Staphoid A-B: toxin based
Lysigin: includes capsular serotypes 5,8, 336, and 5 different SA strains
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| Vaccine Strategies | Elicit antibody responses: capsular components, recombinant proteins/toxins
Limited success: Protein A, low antibody titers in milk
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| Streptococcal Species | Gram + Cocci, catalase negative, some haemolysis
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| Serotyping | Lancefield typing (A-T) of beta hemolytic strep, based on cell surface proteins, use antibodies to recognize surface antigens, group A+B are beta hemolytic
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| Streptococcus agalactiae | Contagious, group B, CAMP positive, obligate of udder, chronic subclinical but can be eradicated, bacteria shed in high numbers, high SCC, 85-95% cure rate with antibiotics, lower duct system
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| CAMP | Strep ag produces a diffusible extracellular compound that in conjunction iwth beta hemolysin of SA causes complete lysis of RBC
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| Environmental Strep | S. uberis, S. dysgalactia, S. faecalis, S. bovis ...... enterococci
Infections during dry period, between milkings
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| Strep Virulence Factors | Polysaccharide capsule, bac protein,, C5a-ase cleaves C5a, Streptolysin S and Streptolysin O, M protein, leukocidins, firbronectin binding proteins adhesions, streptokinase, hyaluronic acid capsule, strep pyrogenic exotoxins, proteases
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| Bac protein | Binds IgA
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| Streptolysin | S: beta hemolysin of group a strep, oxygen stable leukocidin
O: oxygen labile leukocidin
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| M protein | Adhesin, found in fimbriae, blocks binding of complement to peptidoglycan, interferes with phagocytosis, mutations alter structure rendering antibodies ineffective
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| Streptokinase | Cleaves plasminogen to plasmin
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| Strep pyrogenic exotocins | Types a, b, c = superantigens
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| Hyaluronic acid capsule | Strep uberis, no changes in binding of antibody, no effect on macrophage phagocytosis, can lyse/impair function of neutrophils at high concentrations
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| Proteases | Specific nutrient requirements for growth (AA), releases amino acids from microenvironment in mammary gland
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| Strep dysgalactiae | Lancefield serological group C, mouth/vagina/mammary gland skin, dry cows and heifers, 50-60% cure lactation, 85% involution cure, pre dip effective
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| Strep dysgalactiae virulence factors | IgG binding protein, hyaluronidase (digests capsule, inhibits macrophage activity), fibronectin binding protein, fibrinolysin, streptokinase (plasminogen activation)
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| Competence Stimulating Peptide | Mediated quorum sensing system, exponential growth phase = high levels of surface associated proteins, stationary phase = high levels secreted proteins
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| Strep Immune Evasion | Capsule: hyaluronic acid is produced, C5a peptidase: C5a enhances chemotaxis of phagocytes, M protein is fibrillar surface protein, leukocidins, production of toxins, introduce circulating cross reactive antibodies, antigenic variation and disguise
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| Antigenic Variation | Drift: gradual accumulation of minor mutations, decreased recognition by the immune system
Shift: sudden and major change in antigeniciy, failure of the immune system to recognize a new antigenic type
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