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9/9/2015 OBJECTIVES TICKBORNE • Describe the epidemiology, origin, treatment, and prevention of tick borne DISEASES diseases. • Solve case studies. Lynda Britton, Ph.D., MLS(ASCP)CM, SM LSU Health Shreveport TICK LIFE CYCLE BLACKLEGGED TICKS HOW TICKS SPREAD DISEASE MOUTH PARTS • Preparing to feed 10 min to 2 hrs • Cuts skin surface & inserts feeding tube • Secrete saliva to modulate host defenses – Pain and itch – Hemostatis – Inflammation and wound healing • Transmits pathogen while feeding for days • Drops off to complete next life stage 1 9/9/2015 • Emerging Infections Program • CDC in 2007 • Coordinate surveillance – Public health impact –Cost – Tick density and acaridice treatment – Trap rodents and treat with fipronil – Identify novel pathogens molecularly WWW.CDC.GOV/PUBS RESEARCHER DRAGS FOR TICKS CASE 1 • 12 y. o. from Santa Cruz, CA • Swollen painful right knee and hip • Recurrent knee swelling & pain lasting several days every 4-5 months • Treated with herbs instead of antimicrobials • WBC = 7000, HCT 33%, ESR 73mm/h • Knee aspirate: 59,000 WBCs and protein of 5 g/dL • No growth on bacterial cultures but positive by PCR 2 9/9/2015 WHAT DO YOU SUSPECT IS THE LYME DISEASE PATHOGEN? • Most common tick-borne disease in North America 25% 25% 25% 25% • Caused by Borrelia burgdorferi A. Rickettsia • >21,000 cases reported during 2014 rickettsii • 5th among all nationally notifiable conditions B. Borrelia • 90% cases occur in 12 states burgdorferi • Tick vectors: Ixodes scapularis C. Borrelia hermsii I. pacificus D. Ehrlichia sii ri sii chafeensis orfe d herm rickett chafeensis burg ia l Borrelia Rickettsia Borre Ehrlichia I. SCAPULARIS I. PACIFICUS STAGE 1: LOCALIZED LYME DISEASE INFECTION • Erythema migrans • Seasonal (April-September) • Occurs in ~90% • Low risk of disease if tick • Begins 7-10 days after tick attached <36 hrs bite • Signs/symptoms begin 7-10 • Expands over days to weeks days after bite (range 3-30 days) • Central clearing occurs in • May be asymptomatic minority • Rarely fatal • Constitutional: flu-like and lymphadenopathy 3 9/9/2015 DISSEMINATED STAGE DISSEMINATED: • Multiple skin lesions RHEUMATOLOGIC MANIFESTATIONS •Generalized • 60% untreated 20 yrs ago developed joint lymphadenopathy swelling –10% •Malaise, fatigue, – Synovial fluid 24,000 WBC with granulocytes • 5% develop cardiac • Migratory transient intermittent arthritis and effusions involvement • Pain in muscles, bones, – Conductional abnormalities—atrio- tendons, bursa ventricular node block • Baker’s cyst – Myocarditis – Pericarditis DISSEMINATED EARLY NEUROLOGIC LATE NEUROLOGICAL LYME MANIFESTATIONS DISEASE--RARE • Cranial neuropathy • Encephalomyelitis – Bell’s palsy – Parenchymal inflammation of brain and/or spinal • Meningitis-lymphocytic cord – CSF= lymphocytic, mod • Cognitive difficulties ↑ protein, normal glucose • Peripheral neuropathy • Memory, mood, – Intermittent ↓ vibratory sensaon of legs and feet • Sleep disturbances • Encephalopathy – Memory and cognitive impairment – CSF normal HOW IS LYME DISEASE BEST DIAGNOSIS DIAGNOSED? • Characteristic clinical A. Culture in Kelly 25% 25% 25% 25% picture medium • Serology: 2-step B. Histochemical – 1) Polyvalent ELISA: 60- examination of tissue 70% positive by wk 4 C. Peripheral blood – if equivocal or + smear – 2) Western blot (IgG and D. EIA followed by separate IgM): requires 5 Western blot of 10 bands .. .. n. dium at r e smear e in st m m d • PCR lly We exa bloo y Ke b n i ical ed re m u w lt llo • Culture: low yield (27%) che o Cu o Peripheral f ist H EIA 4 9/9/2015 SOUTHERN TICK-ASSOCIATED RASH ILLNESS • Seronegative Lyme Disease • Southern Lyme Disease • “One of the more obtuse diseases. .” SOUTHERN TICK-ASSOCIATED AMBLYOMMA RASH ILLNESS AMERICANUM • Red, expanding-3” • 7 days of bite • Long star tick • Geography • Cause unknown LONE STAR TICK CLINICAL MANIFESTATIONS • Range and abundance ↑over past 20-30 years • Large numbers in Maine and as far west as • Early Lyme Disease central Texas and Oklahoma – Fatigue, headache, fever, and muscle pains • All life stages will feed on humans— • No arthritis, neurological disease or aggressive chronic symptoms • Feed readily on dogs and cats • Saliva can be irritating; redness and discomfort at a bite site does not necessarily indicate an infection 5 9/9/2015 DIAGNOSIS DIFFERENCE WITH LYME DISEASE • Clinically: symptoms, geographical • STARI patients recall tick bite location and tick bite • Time from bite to symptoms shorter • No blood tests because don’t know • Patients with rash less likely to have etiological agent symptoms • Smaller and fewer lesions • More likely to have central clearing CASE #2 WHAT SHOULD BE INCLUDED IN THE DIFFERENTIAL DIAGNOSIS? A. Meningococcemia B. RMSF 13% 13% 13% 13% 13% 13% 13% 13% • In June, 5 y.o. female C. HME • Taken to an ED in Missouri D. Enterovirus • 3-day history of intermittent fever, E. Kawasaki disease headache, mild nausea, and a sore throat F. Drug reactions ° º • Fever of 105 F (40.6 C) G. Streptococcal • Maculopapular rash on her legs, including a F E e n infection i ase v MS HM io R se tions ct bo c a ccem di fe e co rea in h the soles of her feet. t go Enterovirus al f asaki rug o H. All of the above in cc ll n aw D e K A M ptoco re St FURTHER INFO NEEDED CASE CONTINUED • Rash began on the arms and legs same • How long has rash been there? day • Where it appeared before onset of fever? • Did not own a dog • Medications? • No history of recent travel out of the local • Immunocompromising conditions? area • Recent activities? • No history of a tick bite but ticks in the • Ill contacts? area around their house 9 •Travel? • 8.8 x 10 WBC with 5% bands, 70% neutrophils • Tick exposures? • 50 x 109 platelets 6 9/9/2015 ROCKY MOUNTAIN SPOTTED FEVER • Etiologic agent: Rickettsia rickettsii • Vectors: – Dermacentor variabilis (dog tick) – D. andersoni (wood tick) – Amblyomma americanum (lone star tick) RMSF • 91 cases in 2014 • Spring and summer • Incubation period: ~ 7 days (2-14) • Abrupt onset severe HA, F/C, prostration, myalgias, nonproductive cough (later) • GI manifestations very common • CNS: meningitis, meningoencephalitis • Death: 1-2 weeks after symptom onset in 4- 8% (untreated) RASH • Macular rash develops day 2-6 (90%): wrists, ankles, palms, & soles then centrally • Lesions become petechial, hemorrhagic • Vasculitis 7 9/9/2015 WHAT IS THE PREFERRED METHOD DIAGNOSIS TO DIAGNOSE RMSF IN THE LAB? • Clinical & epidemiologic A. Blood culture 25% 25% 25% 25% • Nonspecific lab findings: B. Direct fluorescent – anemia, hyponatremia, low plts, ↑AST antibody • RMSF PCR from rash site C. PCR of rash biopsy is now preferred D. Immunohisto- • IFA: IgM +: 1-2 w but may chemical staining of be false + blood y sy ... • IgG +: 4-6 w iop st culture b d antibod ical t ash r m Bloo f o ‐che escen o • Immunostaining: skin r CR t P is h fluo o t n u m biopsy prior to therapy Direc Im RICKETTSIA PARKERI CASE 3 • Healthy 2 y.o. female southeast Missouri • Mild form of RMSF • Fever 104⁰F, vomitting and diarrhea • Confirmed 2002 • Red maculopapular rash that blanched began on cheek • Amblyomma maculatum tick • 4.9 x 109 WBC, 10.9 g/dL Hgb, 102 x 109 plt • Cattle on Gulf Coast of Texas • Hyponatremia and mildly elevated liver enzymes • Virginia • Inclusion bodies in cytoplasm of monocytes • Antibodies cross react with RMSF • PCR confirmed diagnosis WHAT IS THE SUSPECTED EHRLICHIOSIS PATHOGEN IN THIS CASE? • Obligate intracellular bacteria form A. Anaplasma 25% 25% 25% 25% distinct microcolonies called morulae B. Rickettsia rickettsii • Diseases: C. Ehrlichia • Human Monocytotropic Ehrlichiosis chafeensis • Human Granulotropic Anaplasmosis D. Borrelia hermsii • E. ewingii • “Rocky Mountain Spotless Fever” ii • Ehrlichia muris-like (EML)—MN & WI sis sma ts a een hermsii ricket haf Anapl ia c tts e Borrelia ck Ri Ehrlichia 8 9/9/2015 EHRLICHIOSIS • Southeastern and south-central US • Lone star tick • OK, MO, AR 35% cases • Ehrlichia muris-like in upper midwest • 966 reported case in 2014 SIGNS AND SYMPTOMS BLOOD TRANFUSION AND ORGAN TRANSPLANTATION • Fever, headache • Chills, malaise • Survive for > 1week in refrigerated blood • Muscle pain • Leukoreduced blood • Nausea, diarrhea, anorexia • Infected within 1 mo • Confusion • No cases confirmed by organ • Conjuctival injection transplantation • Rash—more common in children 9 9/9/2015 GENERAL LAB FINDINGS LABORATORY DIAGNOSIS • Serology (IFA) – Negative in first 7-10 days • Anemia – Minimum peak of 1:80 • Leukopenia – Fourfold change • Thrombocytopenia – Positive: week 3 • Elevated AST – Peak: week 6 – Cross-reactivity • Elevated ALT • Morulae detected in 20% • PCR LABORATORY DIAGNOSIS IF MORULAE WERE SEEN IN SEGMENTED NEUTROPHILS, WHAT WOULD BE THE AGENT? • Antibody titer to Ehrlichia chaffeensis 25% 25% 25% 25% antigen by IFA in paired serum samples A. Ehrlichia •PCR chaffeensis • Identification of morulae in leukocytes and B. Anaplasma a positive IFA titer phagocytophilia • Immunostaining of E. chaffeensis antigen C. Rickettsia ricketsii in a biopsy or autopsy sample ii lia D. Borrelia ensis hi p ffe rickets yto cha sia urgdorferi b • Culture from a clinical specimen agoc a burgdorferi chia h li p ickett rreli r a R o Eh sm B la nap A ANAPLASMA PHAGOCYTOPHILUM • Human granulocytotropic ehrlichiosis • Fever, shaking chills • Severe headache • Malaise, myalgia • Nausea, vomiting, diarrhea, anorexia • Cough • Mild self-limiting in most • Fatal if advanced age or immunosuppressed, malignancy or chronic inflammatory • Rash 60% children; 30% adults • More likely to have morulae 10 9/9/2015 • Persons aged >70 and 60–69 years • <1% fatality • Ixodes scapularis and Ixodes pacificus (blacklegged ticks) • New England, North Central and Pacific states LABORATORY DIAGNOSIS EHRLICHIA EWINGII • IgG antibody titer by IFA in paired serum samples • Canine granulocytotropic ehrlichiosis (first week and 2-4 weeks later) • 20 human cases • Positive PCR whole blood in 1st week and •Vector: A. americanum confirmation • First 4 cases of human infection reported • Identification of morulae in leukocytes and a July, 1999 positive IFA titer • Immunocompromised patients from Missouri, • Immunostaining in a biopsy or autopsy sample Oklahoma, and Tennessee • Culture of A.
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  • Ehrlichiosis in Brazil

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    Review Article Rev. Bras. Parasitol. Vet., Jaboticabal, v. 20, n. 1, p. 1-12, jan.-mar. 2011 ISSN 0103-846X (impresso) / ISSN 1984-2961 (eletrônico) Ehrlichiosis in Brazil Erliquiose no Brasil Rafael Felipe da Costa Vieira1; Alexander Welker Biondo2,3; Ana Marcia Sá Guimarães4; Andrea Pires dos Santos4; Rodrigo Pires dos Santos5; Leonardo Hermes Dutra1; Pedro Paulo Vissotto de Paiva Diniz6; Helio Autran de Morais7; Joanne Belle Messick4; Marcelo Bahia Labruna8; Odilon Vidotto1* 1Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina – UEL 2Departamento de Medicina Veterinária, Universidade Federal do Paraná – UFPR 3Department of Veterinary Pathobiology, University of Illinois 4Department of Veterinary Comparative Pathobiology, Purdue University, Lafayette 5Seção de Doenças Infecciosas, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul – UFRGS 6College of Veterinary Medicine, Western University of Health Sciences 7Department of Clinical Sciences, Oregon State University 8Departamento de Medicina Veterinária Preventiva e Saúde Animal, Universidade de São Paulo – USP Received June 21, 2010 Accepted November 3, 2010 Abstract Ehrlichiosis is a disease caused by rickettsial organisms belonging to the genus Ehrlichia. In Brazil, molecular and serological studies have evaluated the occurrence of Ehrlichia species in dogs, cats, wild animals and humans. Ehrlichia canis is the main species found in dogs in Brazil, although E. ewingii infection has been recently suspected in five dogs. Ehrlichia chaffeensis DNA has been detected and characterized in mash deer, whereas E. muris and E. ruminantium have not yet been identified in Brazil. Canine monocytic ehrlichiosis caused by E. canis appears to be highly endemic in several regions of Brazil, however prevalence data are not available for several regions.
  • Antigen Detection Assay for the Diagnosis of Melioidosis

    Antigen Detection Assay for the Diagnosis of Melioidosis

    PI: Title: Antigen Detection assay for the Diagnosis of Melioidosis Received: 12/05/2013 FOA: PA10-124 Council: 05/2014 Competition ID: ADOBE-FORMS-B1 FOA Title: NIAID ADVANCED TECHNOLOGY STTR (NIAID-AT-STTR [R41/R42]) 2 R42 AI102482-03 Dual: Accession Number: 3650491 IPF: 3966401 Organization: INBIOS INTERNATIONAL, INC. Former Number: Department: IRG/SRG: ZRG1 IDM-V (12)B AIDS: N Expedited: N Subtotal Direct Costs Animals: N New Investigator: N (excludes consortium F&A) Humans: Y Early Stage Investigator: N Year 3: Clinical Trial: N Year 4: Current HS Code: E4 Year 5: HESC: N Senior/Key Personnel: Organization: Role Category: Always follow your funding opportunity's instructions for application format. Although this application demonstrates good grantsmanship, time has passed since the grantee applied. The sample may not reflect the latest format or rules. NIAID posts new samples periodically: https://www.niaid.nih.gov/grants-contracts/sample-applications The text of the application is copyrighted. You may use it only for nonprofit educational purposes provided the document remains unchanged and the PI, the grantee organization, and NIAID are credited. Note on Section 508 conformance and accessibility: We have reformatted these samples to improve accessibility for people with disabilities and users of assistive technology. If you have trouble accessing the content, please contact the NIAID Office of Knowledge and Educational Resources at [email protected]. Additions for Review Accepted Publication Accepted manuscript news Post-submission supplemental material. Information about manuscript accepted for publication. OMB Number: 4040-0001 Expiration Date: 06/30/2011 APPLICATION FOR FEDERAL ASSISTANCE 3. DATE RECEIVED BY STATE State Application Identifier SF 424 (R&R) 1.