Professional Documents
Culture Documents
Taxonomy: Order: Spirochaetales Family: Spirochaetaceae Genera: Spirochaeta Cristipira Treponema Borrelia Family: Leptospiraceae Genus: Leptospira
Spirochetes
General Characteristics: i. helical coils (slender, spiral and filamentous organisms) ii. motile due to axial filaments/periplasmic flagella iii.Gram (-), facultative anaerobes iv. Transverse fission v. Special methods of staining: >>Levaditi method for tissues >>Fontana-Tribondeau for smears
Treponema
Treponema pallidum Morphology: i. contains numerous tight, rigid coils which are more or less regular ii. Sluggish motility, drifting motion and flexous movements iii. Obligate anaerobes, microaerophilic iv. Very hard to stain but best observed by darkfield microscopy v. Division time: 30 hours
Cultivation Non-culturable in artificial medium but can be maintained in the laboratory in testicular chancres of rabbits Clinical Infection: Syphilis also known as Lues venerea (disease of blood vessels and of the perivascular areas) >>worldwide distribution in adults and adolescents
Mode of transmission i. sexual contact ii. Direct contact with primary lesions iii. Contact with body fluids or secretions of patients in infections (primary and secondary stages) iv. Prenatal intrauterine infection Types of syphilis: i. acquired: venereal and nonvenereal ii.congenital
Clinical Manifestations:
Stages of syphilis: i. Primary stage: 10-90 days ip typically a single dry lesion, nontender and firm, with a clean surface, raised border and reddish color lesions known as hard chancre or Hunterian chancre appear on the genitalia or within the anal canal systemic signs or symptoms are absent but lymph nodes are frequently enlarged and tender
ii. Secondary stage: 2-10 weeks after 1lesion fever, sore throat, generalized lymphadenopathy, headache and rash lesions be found on other parts of the body (2 lesions called condylomas (condylomata lata, occurs around moist areas like vagina and anus) all secondary lesions are highly infectious
iii. Latent stage no signs and symptoms of active syphilis but remain seroactive
iv. Tertiary stage involvement of the deep organs of the body Examples: gummas = nonprogressive localized lesions of the dermal elements or supporting structures of the body (also known as tertiary lesions (benign tertiary syphilis) ; 3-10 yrs ff last evidence of 2 syphilis
neurosyphilis = involvement of CNS cardiovascular syphilis = commonly involved organs are the great vessels of the heart; 10-40 yrs after 1 syphilis
Congenital syphilis = results from transplacental infection of the developing fetus and is often very severe & mutilating form of the disease
Bejel or Endemic Syphilis = caused by a variant of T. pallidum (T. pallidum subspecies endemicum) = non-venereal form of syphilis that is transported by direct contact (mucous membrane) = Africa and Middle east (arid areas)
Laboratory Diagnosis
>>Specimens: tissue fluid from early surface lesions and blood Methods: a. Direct visualization by darkfield microscopy, FAT, or by special stains of infected tissue like Levaditi Silver Impregnation Technique and Fontana Tribondeau b. Animal inoculation test c. Serologic tests:
c. Serologic tests: i. non-specific or non-treponemal antibody test >> organism is not used as antigen >> a cardiolipin lecithin antigen is used to detect antibody-like ` substance called reagin >>cardiolipin lecithin antigen from deep heart muscle; nonspecific >>reagin nonspecific- can be produced in tuberculosis, leprosy and malaria
Nonspecific flocculation test VDRL Eagle Kahn Mazzini Kline RPR-Rapid Plasma Reagin Hinton USR Untreated Serum Reagin nonspecific complement fixation test Kolmer Wasserman
ii. Specific or treponemal antibody test >>organisms are used as antigen >>used to detect specific treponemal antibodies >>FTA-ABS confirm positive result >>TPI (Treponema Pallidum Immobilization) >>TPA (Treponema Pallidum Agglutination) >>Reiter Protein Complement Fixation >> MHA-TP (Microhemagglutination assay)
Application of methods: a. Primary syphilis darkfield ID provides the most definite and earliest means of diagnosis b. Secondary (STS) serologic tests or treponemal tests for syphilis are almost always (+) c. Tertiary syphilis flourescent test is the most specific test for syphilis Treatment: a. Penicillin drug of choice b. Erythromycin, tetracycline and cephaloridine
Treponema pertenue Morphology: >>closely related to T. pallidum >>serologically and morphologically indistinguishable and are differentiated by the type of lesions produced in experimental animals Clinical Infections: >>Yaws (Frambesia) tropics >>endemic, in children, many humid, hot tropical countries
Mode of transmission: >>direct contact (person to person) other than sexual contact in children under age 15 (skin contact)
Clinical manifestation: i. primary lesion >>mother yaw or Framboise >>painless erythematous papule that heals during subsequent 1 or 2 months
ii. Secondary lesion >>daughter yaw >>resembles the primary lesion; occurs 6 weeks to 3 months later Laboratory Diagnosis: >>no serologic test distinguishable human yaws from syphilis Treatment: >>penicillin drug of choice
Treponema carateum Morphology: >>similar to both T. pallidum and T. pertenue Clinical Infection: >>Pinta-disease of tropical areas of central and south america Mode of transmission: i. person to person contact (skin contact) ii. Rarely by sexual intercourse
Clinical Manifestations: i. primary and secondary flat, erythematous and nonulcerating - the healing lesion first becomes hyperpigmented and later, as scarring occurs, will be depigmented -lesions most commonly occur on the hands, feet & scalp ii. tertiary lesions -uncommon in pinta
Laboratory Diagnosis: >>serologically similar to both T. pallidum and T. pertenue and is distinguished by failure to produce cutaneous lesions in rabbits, hamsters or guinea pigs
Treatment: penicillin
Borrelia
General Morphology and Physiology: Axial fibrils Can be seen with light microscopy Culture: >>microaerophilic >>tickborne borrelia >>28-30 degree C >>generation time: 18 hours all have arthropod vectors
Epidemics usually occur >>in the cold season >>among the crowded and poor >>in homes with inadequate hygiene
A. Relapsing fever (disease) spirochetes of genus Borrelia Morphology: >>helical orgs which are loosely coiled with coarser and irregular spirals >>demonstrated by direct stain (Wright Giemsa)peripheral blood >>microaerophilic and actively motile with a corkscrew-like motion >>11-30 periplasmic flagella
Clinical Infection: >>Borreliosis or relapsing fever (tick fever, famine fever or yellow plague) >>Mode of transmission: i. ticks or human body and head lice ii.transplacental transmission iii.infected blood >>Types of diseases: i. Tickborne or endemic or American Relapsing Fever caused by: -B.duttoni B.anserina -B.turicatae B. hermsii -B.parkeri
Laboratory Diagnosis: >>Specimen: blood (best demonstrated in the course of a febrile period) a. Giemsa and Wright stained blood smears and Gentian violet b. darkfield microscopy b. animal inoculation test c. serologic d. culture using: i. Kellys medium ii. Chick embryo iii.Barbour-Stoenner-Kelly medium iv. Smiberts medium
B. Lyme Disease
illness associated with skin rash, erythema chronicum migrans caused by Borrelia burgdorferi
Morphology: a. resembles other members of the family spirochaetaceae b. microaerophilic c. has 7 to 11 periplasmic flagella
Clinical Infection: Lyme disease (erythema chronicum migrans, Lyme arthritis & Bannwarths syndrome) a. Tick bites b. Vectors (ticks) i. Ixodes pacificus West Coast ii. Ixodes dammini East Coast and Midwest iii. Ixodes ricinus Europe iv. Ambyloma americanum also a vector of tularemia and Rocky Mt. spotted fever
Clinical Manifestation i. first stage >expanding lesion with papule, malaise, fever, headache, stiff neck ii. secondary stage ->onset of neurologic and cardiac involvement ->headache, Bells palsy, radiculoneuropathy, myocarditis, arrythmias
Laboratory Diagnosis >>Specimen: skin biopsy from lesions, synovial tissue, blood, CSF a. stained smears (Dieterles silver stn) b. culture >Barbour-Stoenner-Kelly medium >Mod. Kellys medium c. serologic tests i. ELISA ii. Indirect immunofluorescence assay
Leptospirae
tightly wound spirochetes, about the size of treponemes, shaped like a shepherds crook; saprophytes and animal pathogen only accidentally infecting man
Morphology a. helicoidal, tightly coiled, thin, flexible spirochetes with very fine spirals in which one or both ends is often bent forming a hook b. active rotational motion (corkscrew-like) c. obligate aerobes, oxidase (+), catalase (-) or peroxidase (+)or both d. diaminopimelic acid content of leptospira differentiates from Treponema and Borrelia, which instead contain ornithine
Characterization of Species a. L. interrogans representing all the parasitic strains or pathogenic organisms b. L. biflexa includes all saprophytic strains or water leptospira Clinical Infection: >>Leptospirosis (Swineherds disease, Fort Bragg Fever, Pretibial Fever, Weils disease, Canicola Fever, Autumnal Fever)
a. Mode of transmission: i. direct and indirect contact with infected urine of an animal ii. Ingestion of contaminated food and drinks iii. Infected soil, food and water enter through the mucus membrane or breaks in the skin
b. Clinical manifestation i. infecting serogroups -> severe icteric disease L. icterohemorrhagiae (most common cause of disease) -> less severe & anicteric disease L. australis, L.pyrogenes, -> mild L. canicola, L. ballum,L.pomona
ii. Stages of Leptospirosis (illness is biphasic) - 1st stage variable febrile onset with jaundice, hemorrhage and nitrogen retention - 2nd stage aseptic meningitis with intense headache, stiff neck and pleocytosis in CSF nephritis and hepatitis skin, muscle and eye lesions myocarditis
Laboratory Diagnosis >>Specimens: blood & CSF (1st wk), urine (2nd week) a. darkfield microscopy b. animal inoculation test c. serologic test i. macroscopic slide agglutination ii. Microscopic slide agglutination iii. CFT, FAT, Hemagglutination d. culture i. Fletchers medium ii. Noguchis medium iii. Stewarts medium iv. Ellinghausen, McCollough, Johnson and Harris (EMJH) medium (incubation: dark at RT-114days)