Microbiology BIOL 2230
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Date Created: 10/15/15
BIOL 2230 I REEDER IMIVIUNIZATION Im munization A Immunotherapy Artificial Passive Immunity 1 3 7 Vac 1 2 3 bio22307immunizationdoc First attempts involved the transfusion of horse serum containing antitoxins antibodies to prevent tetanus and to treat patients exposed to diphtheria Today antisera from animals has been replaced with products of human origin due to the stimulation of allergies such as serum sickness or anaphylaxis a Antiserum is a blood serum containing specific antibodies produced against an antigen such as a toxin a virus or a bacterial cell b The horse because of its size and availability was the animal of choice used in such a way so that it could be immunized with either bacteria or their toxic products and then its immune serum used against a disease in a human39 rabbits and cows were also used 1 Today when human immune globulin SIG is not available antisera and antitoxins of animal origin horses can be used diphtheria botulism spider and snake bites Immune serum globulin ISG or gamma globulin contains immunoglobulins extracted from the pooled blood of at least 1000 human donors a Each serum lot contains a broad section of IgG and some IgM antibodies b It is the treatment of choice in preventing measles and hepatitis A and in replacing antibodies in immunodeficient patients Specific immune globulin SIG is derived from a more defined group of donors a SIG serum is derived from patients convalescing and in a hyperimmune state after such infections as pertussis rabies tetanus chicken pox and hepatitis B A major disadvantage is its specificity there are many different antigenic types of pneumococci and meningococci that cause pneumonia and meningitis respectively39 physician must ascertain which antigenic type is the causative agent before the correct antiserum can be administere The major advantage of antiserum is that it provides the patient with circulating antibodies that are effective immediately after its injection passive form of immunity Antiserum therapy a frequently a skin test is performed to determine whether a person is allergic to a particular antiserum by injecting a small amount into the skin and looking for in ammation within a 10 to 15 minute perio Provides temporary immunity in an adult at effective levels for only about 4 to 6 weeks b cination Artificial Active Immunity Conferring active immunity artificially by exposing a person to material that is antigenic but not pathogenic The objective is to obtain the same response as in natural immunity stimulate B and T lymphocytes and create memory clones The exact timing of an immunization sequence depends upon the type of vaccine used the route of inoculation and the immunological competence of the individual Ideal vaccines would stimulate lifelong immunity be completely safe to use require only one administration and be relatively easy to produce a many vaccines in use today do not possess all of these characteristics b for many infectious diseases there are no effective vaccines Most vaccines are injected by subcutaneous intramuscular or intradermal routes a Oral vaccines are available for only three diseases adenovirus infection military recruits poliomyelitis routine childhood and rotavirus newborn infants b Intranasal route for the new in uenza vaccine Tamiflu Relanza Immunity Duration a Realistic expectations the use of a vaccine will be effective and as lasting as the immunity acquired by recovery from the natural disease39 it is not expected to stimulate lifelong protection b Antibodies will reach measurable levels titer within 7 to 10 days after vaccine administration and continue to rise for a week or two and then level off 1 This antibody level may be maintained for 6 months to a year and then usually start to decline 2 Within one to two years at very low levels a second injection of the same vaccine will booster antibody production to high levels within 2 to 3 days Vaccines are grouped according to the antigen type used and the method of preparation killed or inactivated microorganisms living attenuated microorganisms avirulent toxoids inactivated exotoxins or antigenic components and genetically engineered microbes or microbial antigens a Killed inactivated vaccines are prepared by cultivating the desired bacterial or viral strain and treating with formalin radiation or other agent that does not destroy antigenicity 1 Examples include vaccines for pertussis typhoid fever Salk polio and rabies 112408 Page 1 of4 b Living attenuated vaccines are processed to lessen or negate the viral or bacterial in uence 1 Methods include longterm cultivation selection of mutant strains that grow at colder temperatures passage of the microbe through unnatural hosts or tissue culture and removal of virulence genes 2 Examples include vaccines for tuberculosis measles mumps Sabin polio and rubella 3 Live or active viral vaccine provides longer immunity than does inactivated because living organisms also stimulate cellmediated immunity in which inactivated vaccine does not because the living organisms may grow within the individual a much greater antigenic mass can result the continued presence of living organisms continues to stimulate antibody formation there remains the rare possibility that live avirulent strains of virulent organisms may revert to the virulent orm and cause serious disease polio l the oral polio vaccine OPV has been replaced by inactivated polio vaccine IPV for children c Toxins and toxoids purified bacterial exotoxin that has been chemically denatured l Toxins must be treated to reduce their harmful effects forming a toxoid 2 Treated with formaldehyde and prepared by precipitating the toxoid on alum aluminum sulfate which results in a higher level of antibody response 3 Used for active immunization against tetanus and diphtheria d Subcellular or Acellular vaccines are taken from microbial cultures produced by recombinant DNA technology or is synthesized chemically utilizes the exact antigenic determinant that stimulates immunit 1 Examples include capsules of the pneum ococcus and meningococcus the protein surface antigen of anthrax and the surface proteins of hepatitis B Example types of vaccines based on their specific content a Bacterial vaccines 1 Contain either attenuated or killed inactivated bacteria cells a Uses of inactivated strains include actions against various Gram negative related diseases typhoid pertussis plague against common types of Streptococcus pneumoniae against Group A and C meningococci and against tuberculosis b Living attenuated strains tuberculosis tularemia anthrax and brucella in farm animals c Capsular material pneumococcal pneumonia Haemophilus in uenzae and Neisseria meningitidis b Rickettsial vaccines 1 Contain the rickettsial organism grown in yolk sacs of chick embryos or in cell cultures and killed by form aldehyde treatment 2 Not widely used but do offer protection to those people who are more likely to be infected by tick bites 3 Vaccines available against Rocky Mountain spotted fever typhus and other rickettsial diseases c Viral vaccines l Composed of either inactivated or active attenuated virions active vaccines are believed to induce a longer lasting immunity even lifelon 2 Uses include in uenza inactivated all three types of poliomyelitis red measles rubeola German measles rubella yellow fever and mumps which are active vaccines attenuated 9 New vaccine strategies employ antigen synthesis recombinant DNA and gene cloning technology a rDNA technology 1 provides means of isolating the genes that encode various microbial antigens 2 insert them into plasmid vectors 3 clone in appropriate hosts cloning host can be stimulated to synthesize and secrete a protein product antigen which is then harvested and purified 4 Examples hepatitis B rotavirus Lyme disease39 also agents of syphilis Schistosom a and in uenza b quotTrojan Horse vaccine using genetic recombination 1 genetic material from a selected infectious agent is inserted into a live nonpathogenic carrier microbe 2 in theory the carrier microbe will multiply and express the foreign genes and the vaccine recipient will be immunized against the microbial antigens 3 Examples include Vaccinia the original virus used to vaccinate for smallpox and adenoviruses Vaccinia is also used as the carrier in experimental vaccines for AIDS herpes simplex 2 leprosy and tuberculosis c DNA vaccines similar to gene therap l Microbial DNA is inserted into a plasmid vector and inoculated into a human recipient 2 Human cells may take up some of the plasmids and express the microbial DNA in the form of protein 3 Because these proteins are foreign B and T cells will be sensitized and form memor 4 Over 30 DNAbased vaccines are being tested in animals Lyme disease hepatitis C herpes simplex in uenza tuberculosis papillomavirus and malaria The new generation of vaccines last longer reduce the number of booster shots and cover more than four diseases at one time a Already available DPT39 MMR measles mumps rubella chicken pox VZ H u in uenzae 39 hepatitis B bi022307immunizationdoc 112408 Page 2 of 4 amp333 00 L O b New DTP with polio H flu and hepatitis B with DTP or 5 vaccinations at once MMR with chicken pox 11 Vaccines must go through many years of trials in experimental animals and humans before licensing 12 Most complications are local reactions at the injection site fever and allergies C Serological and Immune Tests 1 Serology is the immunological branch that traditionally deals with in vitro serum diagnostic testing a antibodies formed during an immune reaction hold practical value besides combating infection 1 antibody titer or specificity can reveal a patient39s history of contact with microorganisms or other antigens 2 provides a tool for detecting identifying and quantifying antibodies or antigens b Modern serological testing now includes serum urine cerebrospinal fluid whole tissues and saliva 1 These tests are helpful in confirming a suspected diagnosis or in screening a certain population for disease c Serological tests typically exhibit a high degree of specifici pv and sensitivity 1 Specificity focuses only on a certain antibody or antigen 2 Sensitivity can detect even very small amounts of target antibodies 2 Six basic types of serologic tests 1 391 39 39 reaction 39 39 reactions 1 fixation reactions immunofluorescence reactions quot 39 and Wquot m 4 391 39 assays ELISA a Precipitation Reactions The combination of free soluble antigen and antibody molecules forming lattices huge interlocking macromolecular webs three dimensional aggregate reaction is observable in a test tube in which antiserum has been carefully laid over on antigen solution resulting in a cloudy or opaque zone 1 lattices become visible as they precipitate and settle out of solution a precipitation ringtest perform ed in liquid noting a cloudy zone of precipitation where antigen and antibody meet detect syphilis antibodies immunodiffusion assay performed in a gel agar plate containing wells where molecules might diffuse meet and form a precipitate 1 immunoelectrophoresis assay movement of antigen and antibody is accelerated by electrical current 2 Countercurrent immunoelectrophoresis CIE very rapid test for diagnosis of lifethreatening bacterial infections bacterial meningitis and viral a Western Blot for detecting proteins antibody or antigen variation of immunoelectrophoresis 1 Is currently the verification test for HIV antibodypositive people from the ELISA test Agglutination Reactions antigen g antibody is attached to a larger particle such as a cell red blood cell or bacteria or latex bead with determinant groups on the surface when antigen and antibody are combined and insoluble threedimensional aggregate forms and settles out 7 V S7 1 Direct type involves antigens or antibodies that are naturally part of a larger particle microorganisms or erythrocyte a hem agglutination RBCs are mixed with a known virus and a patient39s serum of unknown content 1 Test is based on a competition between the RBCs and the antibodies for the virus antigens 2 Viral diseases diagnosed measles rubella mumps mononucleosis and in uenza b These tests are routinely performed by blood banks to determine ABO and Rh blood types for transfusion preparations c Other examples 1 Rapid Plasma Reagin RPR test for syphilis antibodies 2 Diagnose Mycoplasma pneumonia 3 WeilFelix reaction for diagnosing rickettsial infections 2 Indirect type antigens or antibodies are artificially absorbed onto a latex bead or other particle a Examples include assays pregnancy hormone in the urine identifying Candida yeasts and bacteria staphylococci streptococci gonococci and diagnosing rheumatoid arthritis 3 Tests may be qualitative andor quantitive c Complement Fixation Reaction antibodies bound to antigen can activate complement and initiate the formation of a membrane attack complex that will lyse a target cell an antigen39s or antibody39s presence may be detecte 1 When the antibodies act in conjunction with the intrinsic complement system on RBCs the cells hemolyze lyse and release hemoglobin a uses four components antibody antigen complement from guinea pig blood and sensitized sheep red blood cells b Invaluable in diagnosing certain viral rickettsial fungal and parasitic infections 2 are extremely sensitive and quantitative antibody concentrations can be determined 3 many tedious and complicated steps are involved requiring at least 48 hrs to complete 4 Important verification procedure for group A streptococci infections scarlet fever rheumatic fever and others 5 Also used to diagnose influenza AampB in epidemiology labs Q fever and certain fungal infections p 1 H and 1 d Immunofluorescence Reactions antibodies are tagged with fluorescent dyes that make them easy to identify visually 1 dyes such as fluroescein and rhodamine emit visible light in response to UV radiation bio22307immunizationdoc 112408 Page 3 of 4 BIOL 2230 REEDER UNUSUAL PROCARYOTIC CELLS l Atypical Procaryotes A Rickettsiae sing rickettsia 1 Typically smaller than most bacteria 03 to 07um by 15 to 20um 2 Rod shaped and structurally related to Gram negative bacteria they stain negative with the Gram stain and therefore may be a special type of Gram negative bacteria a contain muramic acid and diaminopimelic acid found only in Gram negative walls in their cell walls b cell wall consists of an inner and outer membrane 3 Features that set rickettsiae apart from normal bacteria similar to viruses in that they are obligate intracellular parasites except for Rochalimaea guintana 39 its pathogenesis also differs involves an arthropod vector a can grow only inside the cells of animals and cannot be grown on artificial media unable to produce sufficient energy to replicate extracellularly b once they are removed from their intracellular environment they lose viability and infectivity39 relates to loss of intracellular metabolites c rickettsiae cell membranes contain unusual transport systems allowing uptake of intracellular NAD CoA and ATP39 normal cells cannot do this d are able to synthesize their own proteins nucleic acids and other macromolecules e are able to oxidize glutamic acid and other Kreb s cycle intermediates to form ATP f none are able to utilize glucose 4 Culturing techniques a infection of susceptible animals such as guinea pigs b grown in tube tissue cultures of living animal cells such as chick embryos l incubate fertile hens39 eggs at 35 C for 710 days 2 organism is then injected into the yolk sac where they can enter and grow in embryonic cells of the yolk sac membrane 3 incubation for an additional 45 days 4 a number of techniques may be used for partially purifying the rickettsia followed by differential centrifugation c for production of certain rickettsial vaccines rickettsiae are grown in monolayer cultures of chick or duck embryo cells 5 Pathogenesis a the manner in which humans are infected differs between rickettsia and other bacteria 1 With one exception Q fever acute disease of animals that can be transmitted to hum ans human infections involve an arthropod vector carrier of pathogenic organism from one host to another such as an infected tick louse ea or mite 2 The disease incidence depends on the geographic distribution of the vector and on the risk of exposure which is enhanced by such things as poor hygienic conditions amp camping in wooded areas b the major reservoir of the rickettsiae is in the wild animal population 1 cycle of infection going from animal to arthropod and back to animal 2 with one exception the hum an body louse which carries epidemic typhus the infected arthropod vector is normally not harmed 3 because rickettsiae are passed along a transovarian route by the vector through the eggs from mother to offspring the wild animal reservoir is not actually necessary for rickettsiae maintenance in nature 4 in those diseases where the vector is not the human body louse humans become infected only when they accidentally enter the anim altoarthropod infective cycle 5 In the USA there are two rickettsial diseases of significance Rocky Mountain spotted fever Bickettsia rickettsii and Q fever Coxiella burnetii 39 other rickettsial diseases such as epidemic Q prowazekii endemic amp scrub typhus are important in developing countries 6 The typical lesion caused by the rickettsiae is a vasculitis particularly in the endothelial lining of the vessel wall where the organism is found characteristic rash edema and hemorrhage due to increased capillary permeability B Chlamydiae 1 Are also obligate intracellular parasites originally referred to as the large viruses are not transmitted using an arthropod vector 2 Have a complex mode of reproduction called the developmental cycle a an extracellular infective elementag body small dense cell about 03um in diameter is taken into the host cell through the cell membrane BlO2230 l l l SUALPROCARYOTIC 102204 an initial or reticulate body then forms from the elementary body reorganization into a large metabolically active dense cell c large cell grows and multiplies fission d after 2448 hours the reticulate bodies noninfectious reorganize into small dense infective elementary bodies once again e host cell bursts releasing these infective cells 3 believed descendants of Gram negative bacteria 4 parasite feature relates to their inability to synthesize their own ATP which is required to be metabolically complete a have unusual transport systems which allow for ATP and as well as large protein molecules 5 Their complex developmental cycle distinguishes them from all other procaryotes a no other bacterial organism produces daughter cells that must undergo a morphological reorganization before they become infective 6 Divided into two subgroups based on the morphological appearance of their intracellular inclusion bodies presence or absence of glycogen in their inclusions their sensitivity to sulfonamides and the extent of DNA homology between related organisms a b Group A form compact inclusion bodies containing glycogen and are inhibited by sulfonamides causative agents of trachoma inclusion conjunctivitis nongonococcal urethritis epidymitis female pe vic in ammatory disease and lymphogranuloma l Trachoma occurs only in humans chronic in ammation of the conjunctiva can lead to blindness transmitted by direct contact with fingers or contaminated towels clothing and sexual transmission Group B different inclusion body with no glycogen and is resistant to sulfonamides psittacosis infects the lungs primarily causing either asymptomatic features rising antibody titer g high fever amp pneumonia atypical l C psittaci infects birds amp mammals humans are infected primarily by inhaling organisms in dry bird eces 2 C pneum oniae atypical pneumonia described by pharyngitis bronchitis and pneum onitis C Mycoplasmas l Smallest organisms known capable of growth and reproduction outside living host cells a variable sizes averaging 012 to 025um diameter 2 Major morphological difference from other procaryotes lack a cell wall a my in size and shape due to exible 3layered cell membrane ultra microscopic coccoid cells to branching or unbranched long filaments 3 Theirs is the only bacterial membrane that contains cholesterol normally found in eucaryotic cell membranes 4 Reproduction is still unclear a coccoid cells routinely elongate into a long filament followed by formation of coccoid structures internally b fragmentation will subsequently result in the coccoid structure release c budding may also occur 5 Vary in their growth requirements but all can be grown on artificial media a growth on solid media results in colonies too small to be seen with the naked eye 1 under low power colonies show a quotfriedegg look 2 T strains when grown are even smaller and require both cholesterol and urea for growth 6 Widespread in humans and other animals a causative agents of pleuropneum onia in cattle arthritis in rats and rolling disease neurological disorder in mice b found commonly in the human mouth and frequently in the human genital tract especially T strains 1 in the genital tract T strains may cause in ammation c found isolated from the joints of arthritis patients has not been proven as to its involvement in the arthritic syndrome d causative agent of atypical pneumonia in hum ans with increased incidence in the winter M pneumoniae a pathogen only for humans is transmitted by respiratory droplets respiratory mucosa is not invaded but ciliary motion is inhibited and necrosis of the epithelium occurs 2 quotAtypicalquot means that a causetive bacterium cannot be isolated on routine media or that the disease does not resemble pneumococcal pneumonia e associated with many plant disorders D L Forms 1 Derivative forms found from the normal structural appearance of different bacterial types smaller sized cells lacking cell walls BlO2230 l l l SUALPROCARYOTIC 102204 BIOL 2230 Reeder Hyperclinic Report Layout Organismg s1 Causes of Disease s A General Description Pathogenesis and Epidemiology of speci c disease 1 General description 2 Pathogenesis include virulence factors of causative agents if required viral bacterial etc 3 Epidemiology General Description Pathogenesis and Epidemiology of speci c disease IF assigned more than one disease 1 General description 2 Pathogenesis 3 Epidemiology Body Systems Involved 1 system is comprised of a Include speci c parts impacted by the disease or infection Example Pathogenesis and Epidemiology of Meningococcal Disease B BIOL2230 HyperclinicReportLayout Pathogenesis a upon reaching its portal of entry in the nasopharynX pili of the meningococci allow it to attach this may result in simple asymptomatic colonization in some individuals the meningococci are engulfed by mucosal epithelial cells and penetrate nearby blood vessels damages epithelium and causes pharyngitis in the brain via the blood vessels the bacteria rapidly cross the bloodbrain barrier permeate the meninges and produce the following symptoms fever sore throat headache stiff neck convulsions and vomiting meningococcemia can also occur as the pathogen sheds endotoxin into the generalized circulation providing a potent stimulus for certain leukocytes release of cytokines by the leukocytes damages the blood vessels leading to vascular collapse hemorrhage and lesions called petechiae on the appendages and trunk in some cases meningococcemia progresses and can lead to death sudden onset of fever higher than 40 C chills delirium widespread ecchymoses shock and coma cardiac failure generalized intravascular clotting damage to the adrenal glands and death within a few hours miology have a sporadic or epidemic incidence in the late winter or early spring g Epide a Page 1 of3 11272007 II III BIOL2230 HyperclinicReportLayout human carriers in their nasopharynX represent the continuing reservoir of infection the carrier state can last from a few days to several months in 3 to 30 of the adult population and can exceed 50 in institutional settings transmission is set when carriers live in close quarters with nonimmune individuals as in college dormitories daycare facilities and military barracks highest risk groups are young children 636 months old and older children and young adults 1020 years old Neisseria meningitidis is the second most common cause of meningitis in adults after Streptococcus pneumoniae Pathogens Responsible A Bacterial causes of disease l Microbial characteristics a Organism name 1 Description 2 Habitat 3 Transmission 4 Incidence 5 Virulence 6 Detection b B Viral causes of disease Microbial characteristics a Organism name 2 3 etc b Case Study Description Initial Prognosis and Potential Causes 1 Case descriptions 2 Your prognosis relating case description and information that you have researched thus far 3 Bacterial causes of a list organisms b c 4 Viral causes of a list organisms c 5 Fungal causes of Page 2 of3 11272007 BIOL 2230 Reeder Hyperclinic Report Layout Organismg s1 Causes of Disease s A General Description Pathogenesis and Epidemiology of speci c disease 1 General description 2 Pathogenesis include virulence factors of causative agents if required viral bacterial etc 3 Epidemiology General Description Pathogenesis and Epidemiology of speci c disease IF assigned more than one disease 1 General description 2 Pathogenesis 3 Epidemiology Body Systems Involved 1 system is comprised of a Include speci c parts impacted by the disease or infection Example Pathogenesis and Epidemiology of Meningococcal Disease B BIOL2230 HyperclinicReportLayout Pathogenesis a upon reaching its portal of entry in the nasopharynX pili of the meningococci allow it to attach this may result in simple asymptomatic colonization in some individuals the meningococci are engulfed by mucosal epithelial cells and penetrate nearby blood vessels damages epithelium and causes pharyngitis in the brain via the blood vessels the bacteria rapidly cross the bloodbrain barrier permeate the meninges and produce the following symptoms fever sore throat headache stiff neck convulsions and vomiting meningococcemia can also occur as the pathogen sheds endotoxin into the generalized circulation providing a potent stimulus for certain leukocytes release of cytokines by the leukocytes damages the blood vessels leading to vascular collapse hemorrhage and lesions called petechiae on the appendages and trunk in some cases meningococcemia progresses and can lead to death sudden onset of fever higher than 40 C chills delirium widespread ecchymoses shock and coma cardiac failure generalized intravascular clotting damage to the adrenal glands and death within a few hours miology have a sporadic or epidemic incidence in the late winter or early spring g Epide a Page 1 of3 11272007 II III BIOL2230 HyperclinicReportLayout human carriers in their nasopharynX represent the continuing reservoir of infection the carrier state can last from a few days to several months in 3 to 30 of the adult population and can exceed 50 in institutional settings transmission is set when carriers live in close quarters with nonimmune individuals as in college dormitories daycare facilities and military barracks highest risk groups are young children 636 months old and older children and young adults 1020 years old Neisseria meningitidis is the second most common cause of meningitis in adults after Streptococcus pneumoniae Pathogens Responsible A Bacterial causes of disease l Microbial characteristics a Organism name 1 Description 2 Habitat 3 Transmission 4 Incidence 5 Virulence 6 Detection b B Viral causes of disease Microbial characteristics a Organism name 2 3 etc b Case Study Description Initial Prognosis and Potential Causes 1 Case descriptions 2 Your prognosis relating case description and information that you have researched thus far 3 Bacterial causes of a list organisms b c 4 Viral causes of a list organisms c 5 Fungal causes of Page 2 of3 11272007
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