VERVE eee ee lit ee "AL PARASITOLOGY Coccidians OUTLINE + thin-walled oocysts - infect other enterocytes thus |. ntroduetion resultng in autanecion responsible forte conic of ane the infection among the immunocompromised) B._ Stages of Coccidians |. Cryptosperiaium hominis ll. Cyclospora cayetanonsis , IV. Gytoisospora bel V. Cryptosporidium parvum Vi. Toxoplasma gondii Vii. Sarcocystis spp. ‘A. Description Largest group of apicomplexan protozoa Class: Conoidasida Subclass: Coccidia Spore-forming, single-celled obligate intracellular protozoan ‘Infect the intestinal tract Causes coccidiosis B. Stages of Coccidians 41, Sexual cycle or sporogony - producing oocysts 2. Asexual cycle or schizogony (merogony) - producing merozoites (meronts), 3, Gametogony - resulting in the development of male (micro) and female (macro) gametocytes (gamonts. Ene ‘A. Introduction ‘All stages of development are completed in the gastrointestinal tract of the host ‘= Water-bome transmission + Chlorination does not affect the parasite + Infective stage: Oocysts B. Morphological forms 1. _Oooytsts to5 um Each contains four sporozoites (present at time of passage into the feces) Infectious when ingested Sporozoites — small trophozoites — intracellular but ‘extracyloplasmic that attaches to brush borders "+ Trophozoites divide by schizogony 3._Micro and macro gametocytes ae ‘= Zygote - macrogamete is fertilized by the microgamete + Two types of oocysts resulting from a zygote * thick-walled oocycts - passed out with the * feces that may contaminate food and water . Pathogenesis & Clinical Manifestations: 1. Cryptosporidiosis, = Selflimiting diarrhea Tasting for 2 to 8 weeks ' abdominal pain, anorexia, fever, nausea, and weight los 2._Acute and gangrenous cholecystitis, * Bile duct and gall Bladder 3._Respiratory infections ‘+ Tead to chronic coughing, dyspnea, bronchiolis, and pneumonia, 4, Blunted vil of intestines * infltration of inflammatory cells into the lamina propria and ‘elongated crypts ‘+ malabsorption and excessive fluid loss D. Laboratory Diagnosis 4. Sheather's sugar flotation and the formalin etherfethy! acetate concentration technique - common 2. Acid-fast staining - quickest and cheapest 3. Kinyour's modified acid-fast stain - oocysts appearing as red-pink doughnut-shaped circular organisms in blue background 4. Blopsy - microvillus region of the infected enterocyte 5. Sputum - pulmonary infections 6, Transbronchial and broncheo-alveolar lavage - pulmonary infections 7. Indirect fluorescent antibody, enzyme immunoassay, ‘and DNA probes specific for C. hominis E. Treatment = Nitazoxanide * Bovine colostrum, paromomycin and clarithromycin = diarhea = Azithromycin * Chemotherapy, body fluid replacement and symptomatic treatment - immunocompetent and immunosuppressed Patients PAPER PIPER PH |. com/esperpicerioh 4‘A. Introduction ‘© Photosynthesizing organelles and autofluorescing panicles characteristic ofthe blue green algae ‘Also known as cyenobacterium-like body (CLB) athogeneciy is the same with Crytosporidiasis Infective stage: Oocyst ‘Mode of Transmission: Ingestion of mature oocyst Infected patients shed oocyst: form two sporooysts each ‘containing only two sporozoites Habitat: jejunum + Fission of sporozoites inside the cells -> meronts (contain 8 to 12 merozoites on 1% gen and 4 merazoites in 2% gen) —+ some merozoites develop into male (micro) and female (macro) gametocytes —+ microgametes fertlize the _macrogametes to produce oocysts > passed out in feces. + oocysts undergo complete sporulation within 7 to 12 days in warm environment ‘Number of ‘Contents of sporocysis | Each sparocyst contains wo sporozoites, . Life Cycle @DPDx Cyclospora cayetanensis D. Pathogenesis & Clinical Manifestations Cyclosporiiosis Symptoms: malaise and low grade fever 12 to 24 hours after exposure; fatigue, anorexia, weightloss, nausea, vomiting, abdominal pain, ftulence, bloating, and dyspnea ‘© Chronic and intermittent watery diarhea ‘© early in infection ‘© may continue for 6 to 7 weeks with six or more stools per day + D-xylose malabsorption E. Laboratory Diagnosis 1. DFS in 400x (hing) magnification - recommended 2. Concentration technique - wio the use of formalin ‘ixative, with the addition of 5% Potassium dichromate 3. Kinyoun stain acid-fast 4. Fluorescent microscopy - blue or green oocysts (365- 450 DM) 5. Satranin Stain and microwave heating 8. PCR - to differentiate with Eimeria species F._Treatment ‘© Trimethoprim-sultamethoxazole - only ifit is needed ‘© 160/800 mg twice dally for? days PAPERPIPERPH | com/osperpivereh anareeeeneeaeenaeeaaneeaaas e € e € € € Seen eaewewewvwevevweveeeevvwvw © orn ann ran a ETE TOTO Oe ‘A. Introduction + Infective stage: Oocyst ‘+ Mode of Transmission: Ingestion of oocyst © Contains 2 sporoblasts (immature) © contains 2 sporocyst (mature) with 4 sporozoites each © B sporozoites in all = Common to children and male homosexuals with AIDS ‘Ingestion — sporozoites excyst in small intestine releasing sporozoites penetrating epithelial cells» starting asexual stage or schizogonic phase + Sporozoites to schizont —+ ruptures in the epithelial cell liberates merozoites in the lumen ‘= Some of the merozoites undergo gametogony to produce ‘macrogametes and microgametes (sexual stages)— form ‘zygote that eventually matures — form an unsporulated oocyst. ‘= Sporulation - within 48 hours after passage with the stool ‘Shape ‘and Transparent Developing sporoblast | Unicellular with granular cytoplasm Membrane “Two layered Young oocyst ("Two sporoblasts Mature oocyst “Two sporocysts, each containing four sausage-shaped sporozoites ._Life cycle oe — © ee A Ope ee Above ne \ Ge e2eoo 28 0So Aa 9002 —n © D. Pathogenesis & Clinical Manifestation Cystoisospariasis * generally asymptomatic or may present as a self-limiting {gastroenterits + Severe: diarthea and fat malabsorption Symptoms: Symptoms include low-grade fever, anorexia, \oriting, general body malaise, weight loss, and flatulence. Stoots: undigested food, mucus, and Charcol- Leyden crystals «Inflation ofthe lamina propria with lymphocytes, plasma calls, and eosinophils E. Laboratory Diagnosis 1. Direct Microscopy - Charcot Leyden crystals 2. Concentration techrique: FECT, ZnSO4, and Sugar Floatation Technique 3. Staining techniques: Phenol-auramine, Iodine, Kinyoun, ‘Auzamine-Rhodamine, modified ZiehkNeelsen (red ‘olor against a green background) 4, Enlorotest and duodenal contents 5. Molecular Testing 6. Blood examination - peripheral eosinophilia F. Treatment ‘+ Asymptomatic - bed rest and bland diet, ‘= Trimethoprim-Sulfemethoxazole (SXT) ‘© 160/800 mg four times per day for 10 days, then ‘two times per day for 3 weeks ‘+ Pyrimethamine and sulfadiazine for 7 weeks PAPER PIPER PH | A.com/ogperpioerphAM idlum (093 sponte ~ ‘Size range 46m ‘Shape Roundish ‘Number of sporoaysts ‘None ‘Number of sporozoites | Four, small ‘Schizonis, Four to eight merozoites ‘Micro and 24 um ‘A. Introduction ‘occurs only in the members ofthe cat family (Felidae) intracellular infects different kinds of nucleated cells including macrophages Definitive Hosts: Cat Felidae) Extraintestinal stages (asexual): tachyzoites and bradyzoites Infecive stage: tachyzolte, the bradyzoite, and the oocyst Habitat: Intestinal epithelium ‘© Merozoites mullyply (schizogony) —> differentiate into microgametocytes and macrogametocytes (gametagony) ~> fetlization ofthe macrogamete by the microgamete — oocysts ooysts - passed out with the feces ofthe cat in the Lunsporulated stage and complete sporulation within 3 to 4 ‘© Inside the mature oocyst two sporocysts are formed ‘© each having four sporozoites: When the mature oocyst reaches the intestine of the new host, it excysts and releases four sporozoites which can penetrate the lamina propria —+ gain entry to the Iymphatics — other organs “Tachyzoites - found during the inal and acute stage ‘ofthe infection ‘© can be transferred from the newly infected mother to the fetus during the first trimester Reproduction: asexual multiplication by variation of binary fission or endodyogeny ‘© formation ofthe plasma membrane by the two new ‘daughter parasites, even before the division of the nucleus Pseudocysts containing proliferating tachyzoltes - seen in tissue sections taken from patients suffering from acute infection Cysts - found in muscles and CNS B._Morphologial forms Toxoplasma gondil Oocyst “s ae Sia ange 10-1682 Shape Round to siahy ova . Pathogenesis & Ctnicl Manifestation ‘Number a sparagas —[ Two, Grpesporisiass Number of sporeztes | Four * nfctve sioge:Goayat Membrane Bordad by Gear, colores 5 Mode of Transmission: Ingeton of mature oocyst {wo layered thin cl wal + Diathoa 5 Malabsorption Toxoplasma gondi Tachyzoes D. Laboratory Diagnosis 4. Sheathers sugar floatation technique ay 2 Riyun mode ect fast st sonst npear sed (ow? pin doughnut shaped oredr organisms — 2. k«raminrteae ara) Qe 4 DNApebe Sie ene BTx2a pm E._Troatmont Shape Grescen-soped © spam & Nitozoxeide Generl comment ‘ately mulling 5 Bovine colostrum, Paromycn& Clrtomych rmorphelane fo {Nimiber ofr One PAPERPIPER PH | fé:con/oapernicerph - i nee ek oe Oe @ & & Ob OG © & © OO O OO OO 6 6.88 8 28 oes fe oooVUUVVVE VV VUVVVVVVVVUUVUVUVUVVUVUVUVUEVUUVEEYUUUYS [Toxoplasma gondii Bradyzoites 8 w | - | Sizerange | Smaller than tachyzores | | General comment Slow-growing morphologie form ophozoite ‘ ‘crescent-shaped pointed anterior rounded posterior end ‘hoptries and micronemes - organelles found in the conoid {or cell penetration ‘+ Nucleus: spherical, posterior ©. Life cycte Oseteetsesone oe Sem te J epee oo Nee ee D. Pathogenesis & Clinical Manifestation Toxoplasmosis, ‘Commonly asymptomatic i immune system is good '= Most common manifestation: encephalitis ‘+ Other manifestations: Myocarditis and focal pneumonia, retinachoroiditi, lymphoreticular hyperplasia with enlargement of the posterior cervical lymph node, hepatitis, splenomegaly, pneumonia, extramedullary hematopoiesis, and failure to gain weight. + Stibith and abortion in first trimester Babies - chorioretints, epileptic seizures, jaundice, hhydrocephaly, and microcephaly. Laboratory Diagnosis Examination of Ussue imprints stained with Giemsa Biopsy: stained with H and E ‘Serodiagnostic method: positive titer or a four fold rise ‘Sabin Feldman Methylene blue dye - maintenance of live organisms in the laboratory 5. KAT tot 6. IFAT ora double sandwich IgM enzyme immunoassay - high titers (1,024), although usuell indicating an acute infection, may algo be seen in chronic cases, hence the need for IgM antibody detection 7. Hemagglutnation test 8. Latox agglutination test 8. ELISA 10. PCR serum, amniotic fuid, cerebrospinal ui, and broncheoalveolar lavage F. Treatment +” Pysimethamine (25-100 mg daly) and Sulfadiazine (11.6 {four tmes daly) used in combination © Keep the Toxoplasma under control but do not kl it © Pyrimethamine - can lower blood counts in most people, given together with leucovorin (Folic acid) © Sulfadiazine - cause serious allergic reactions like fever and rash, can be substituted with cingamycin Spiramyoin: pregnant ‘Azithromycin, clarithromycin, dapsone, and atovaquone + Corticostereis - prevent occurrence of hypersensitivity reactions + Trimethoprim-sufamethoxazole - prophylaxis PAPER PIPER PH |. @com/poperpiperoh 53‘A. Introduction * Causes sarcosporiiosis or sarcocystosis. * Birds, reptiles, mammals, and snakes * Cysts - white threadlke in striated muscles of a house mouse ‘+ Mieschers tubules ‘= Protozoa or fungi! but now a protozoa ‘= Definitive hosts: Humans ‘+ Intermediate host: Cow or pigs ‘= Definitve host eating intermediate hosts - undergoes. sexual reproduction within the intestines © Type 10 cell walls Species 1. Sarcoaystis hominis - sitiated fom vilar protrusions 2. Sarcocystis suihominis- with vilar protrusions B. Morphological forms 4. Zoite ‘> simplest form ‘+ banana-shaped cell, with @ pointed anterior end (apical complex) micronemes, micropores, and thoptres - penetration and Creation ofan intracellular environment Sporulated oocysts and individual sporocysts passed out in the feces ‘capable of surviving on the ground sporulated oocyst undergoes sporogony creating two ‘sporocysts ‘* Once sporogony is complete ~> oocyst undergoes lysis ~> releasing the sporocysts into the environment + contain four sporozoites and a discrete refractile residual body * Ingestion of oocysts/sporocysts —+ sporooysts pass to the smal intestine ‘= Schizogony or merogony - lasts about 15 to 16 days 3._Merozoites * emerge from the Second generation meronis| ‘= enter the mononucleate cells ‘© develop in the direction of blood flow to arterioles, capillaries, venules, and veins throughout the body Schizonts * third asexuel generation ‘+ form metrocytes and encyst in the musctes, initiating ssarcocyst formation 5. Sarcocysts “© agin as unicellular bodies containing a single metrocyte 6._Bradyzoites * As sarcooysis mature, crescent-shaped bodies = sarcocysts are ingested and the wall is digested, bradyzoites become motile + enter intestinal cells and change into the male and female forms, microgamonts and macrogamonts, 7._Zygote_ * fusion of @ macrogamont and a microgamont * develops into an oocyst (containing two sporocysts) ‘Infected by consumption of uncooked or undercooked meat of intermediate host that contains sarcocysts. E. Pathologie & Clinical Manifestations. 4. Sarcosporodiosis ‘a Invasive form ‘= Presents with vascultis and myositis * involve tissues including lymph nodes, muscles, and the larynx ‘= Symptoms: anorexia, nausea, abdominal pain, distension, diarrhea, vomiting, dyspnea, and tachycardia ‘= Transient and lasted about 36 hours Intestinal form Nausea, abdominal pain, and diarrhea rormally mild and lasting under 48 hours may occasionally be severe or even life threatening 2. Sarcocystosis ** Associated with acute fever, myalgias, bronchospasm, pruritic rashes, lymphadenopathy, subcutaneous nodules ‘with concurrent eosinophilia, elevated erythrocyte sedimentation rale, and elevated creatine kinase levels ‘Segmental necrotizing enteritis * Symptoms may last as long as 5 years. Laboratory Diagnosis Presumptive: hisiory and sympioms ‘Stool examination - sporocysts in feces ‘© S. hominis excreted 14 to 18 days (beef) © S. suihominis excreted 1 to 13 days (pork) PAPER PIPERPH |. fb com/oaperpiverph g DrAmPR> PSF AWDBDADPAFADARIFAHAADRSHAHMVMIA*HSHMMADPMSPKKRKRHPRKRRRRAAAASae ee a a oe ee ee a ee 3. Fecal flotation wet mount - visualize sporocysts using bright-eld microscopy (© Flotation methods based on high-density solutions incorporating sodium chloride, cesium chloride, zine sulfate, sucrose, Percoll, Fico Hypaque, and other density gradient media - preferred over FECT 4. Microscopy - cannot distingush species 5. Hematoxylin and eosin stain - sarcocysts ‘© Periodic acid-Schiff (PAS) - confirmatory staining 6 Biopsy - definitive diagnosis, . © S. hominis - microscopic in muscles of cattle © S. suihominis - macroscopic in muscles of swine 7. PCR~ 188 rRNA, stored in potassium dichromate {(KeCr207) for as long as 6 years G. Treatment = Asymptomatic * Albendazole, metronidazole, and co-trimoxazole - for myositis * Corticosteroids - for symptomatic rele PAPER PIPER PH |. fcon/naperpiperphCLINICAL PARASITOLOGY 0. Other Intestinal Protozoa OUTLINE 4._Mutple fission _ |. Blastocystis hominis * arise irom vacuolated forme ‘A. Introduction + prominent and tick, osmophili, electron dense wall 8. Morphological forms + sharply demarcated polymorphic, but mosly oval or Pathogenesis & Cical Manifestations circular, dense body surrounded by a loose outer D. Laboratory Diagnosis . membranous layer E. Troatment + membranous layer coresponds to the firilar layer - I Dientamoeba fragiis easiest ciagnoste feature to identiy ‘A. Introduction + thickewalled ost - may be responsible for extemal 3. Morphological forms transmission ©. Mode of Transmission D. Pathogenesis & Cinical Manifestation ‘Blastocystis hominis vacuolated form E. Laboratory Diagnosis E ee F. Treatment e Introduction \ — * previously classified as yeast under the genus \ Schizosaccharomyces @ + related to Blastomyces based on its glistening : Appearance in a wel mount and the absence of any organelle of locomotion + Lacks @ cel wall = = + possesses null endoplasmic ecu, Goi complex, Saeranae Sn {and mitochondriomike organelles Conn ted + capable of pseudopodtal extension and retraction Se ee * does not grow on fungal culture media eeperlae + Reproduction: binary fission or sporulation under strict Atboare 35 Ang around ae Sa periphery of organism + Optimal lea Number of nudlet “Two to four located in ‘© Stramenophiles - small subunit FRNA (SSUrRNA) gene cytoplasm | + Schizosaccharomyces + Blaston Clit a eee C. Pathogenesis & Clinical Manifestations ‘© listening appearance Tn Elesunyeosie © Nolocomotor organelle * Symptons: bdominal anos, ale bowel nde bloating, flatulence, mid to moderate diarmhea without 8. Morphological forms foal odkecyts rood, nausea, voming,ew grade {Nee fever, and malaise ‘= most predominant forms in fecal specimens «= last about 3 to 10 days, but may sometimes persist for + spherical in shape veo ornate nee oe + large central vacuole * four nucle! to the periphery ofthe cet ._ Laboratory Diagnosis * very thick capsule 1 Muttple stool samples with signs and symptoms + prominent central vacuole - reproductive organelle 2. DFS~leukooytes and eosinophilia + main ype of Blastocystis that cause diarrhea 3. Concentration techniques - sensitive 4. Hematoxylin or trichrome staining -dfferentit various 2 _Amesbe tee __ stages * exhibit active extension and retraction of pseudopodia 5. Culture - Boeck and Drbohia’s or the Nelson and Jones. ‘= nuclear chromatin - shows peripheral clumping ‘media * intermediate stage between the vacuolar form and the precystic form maaristnaneiacoseea + predominated in isolates Drug of choice: Metronidazole predominated in isolates from symptomatic cases SE din 3._ Granular © Pediatric dose: 35-50 mg/kg/day in tree doses for ‘multinucleated — S days + lodoquino! ‘© 650 mg three times daily for 20 days ‘= Trimethroprim-sulfamethoxazole (TMP-SMX) ‘= Nitazoxanide - release symptoms 3 days after ‘administration ‘= mainly observed from old cultures * develop into daughter cells of the ameba-form when the coll ruptures PAPER PIPER PH | fb com/oacersiperh 87 PEUVUPREVUVUDUDVUVEVUVYDUDUEOVUVUVeUUEeeaeedsefe Denarcesaagis | ee mabe fopelite + AUDINL sea anpt of he lage rine (anaonaet and outermost) + Nope togs ented 2 eee cases niente Sewers tard tomrties Michomehas « acreeie ny intense sheeted tai Seana mates oenaauraams nocan wecannine mesoee . B._Morphological forms. Dientemoeba Trophozoite | ude —@ Novonatn games rar 88 ‘Size range 7-12 pen Motility Shape Tregular ‘Number of nudiet ne or two (rarely tree ar ‘Appearance | = Karyosome Four to si chromatin - Z discrete granules Undulaing Membrane | Membrane ytoplasm Flagela Other structures ‘Vacuoles with ingested debris ie 1 ©. Mode of Transmission + Ingestion of trophozoites owas Vain’ ‘Peran'a parson \ Dientemoeba fragilis, D. Pathogenesis & Clinical Manifestation, ‘= Excess mucus & hypermotity of feces ‘© peristalsis - movement from large intestine to Fectu ‘+ Loss of appetite, colicky, abdominal pain & intermittent diarrhea + Intermittent diarrhea: Excess mucus, abdominal tendemess, bloating sensation & flatulence ‘= Anal pruritus (11 %)- itching of bottoms, coinfection with Enterobius ‘© Chronic infection: Mimic Imitable Bowel Syndrome (IBS) E, Laboratory Diagnosis 1. Observation of binucleate trophozoites in mutipe feed and 2. slained fresh stool samples. ‘8. Purged stool specimens - provide more suitable material 4. Concentration methods - don't work ‘5. Polyvinyl alcohol fixative or Schaudinn’s fixative F. Treatment = lodoquinot ‘© 650 mg three times daily for 20 days. © 40 malkgiday in three doses, also for 20 days = Totracyctine & metronidazole PAPERPIPER PH | fbcom/asperpiberph s See HPSPPHPHOHH HHH HOH HSH HHH HPHKRKSSRSHSSRSHRASRARARHRAARPROV VOOVVVEVEDVVDUOEEES EVDO es VUYUUUEOY CUNICAL PARASITOLOGY Sporozoa OUTLINE |. Plasmoalum spp. Plasmodium knowles! Jil, Babesia spp. ‘A Introduction Malaria “mal meaning = “aria” meaning air Definitive Host: Mosquito Infective stage of human: Sporozoites, Infectve stage of vector: Gametocytes Leading parasitic disease that causes mortality worldwide, ‘Milennium Development Goals (MDGs) MDG 6 aims to reduce the burden of HIVIAIDS, malaria, ‘and other diseases. ‘Malaria component: reducing incidence and mortality ‘ates , increasing insecticide-reated bed net coverage ‘among children below 5 years of age and increasing ant- ‘malarial coverage among children below 5 years of age «= Infective stage: sporozoites Vectors ‘1. female anopheles minimus var. flavirostis - also known {as the tiger mosquito because of its size and stripes black and white Anopheles litoralis ‘Anopheles maculates ‘Anopheles mangyamus ‘Anopheles balabacensis Species 1. Plasmodium faleiparum 2 Pvivax 3. P.ovale 4. P. malariae 5. _P. knowles! a parasite of iong-talled macaques (Macaca fasciculars) Reproduction ‘Asexual * Schizogony- formation of merozoites = Gametogony - formation of gametocytes Occurs in human Semel ‘= Sporogony - formation of sporozoites ‘= Occurs in mosquito Plasmodium Plasmostum Plasmodium Plasmodium vier ovale ‘malariae Single or mule rng 2._Trophozoite (band form) Plasmodium Plasmodium Plasmodium Plasmodium wor ‘ovale ‘malariae falciparum amoeboid fimbriated ‘erm Plosmodium "210.24 @ “Bio 12 “81036, Inrosette frm merozoites 4. Microgametocytes Pasnodum Plsmexhum Plasnedim Plasmodium ao eae mbes Ogee Maciipwietcca Plesmodtum Plesmodum ‘war ovale (a Plasmodium Plast maloroo faleparum PAPERPIPERPH | f:con/ooperpiperphPavameia | Plasmodium | Piasmodi | Pisamodi | Plasmodl | falciparum | umviver | "um | umovale malarie Nomal | Enlarge | Normal | Enlarge d é ‘Single, | Single | Single | Single Mattie 2 Single, | Single, | Single, | Single, double | Dense, | double | double Big Present | Absent | ‘Absent | | Usually not | Ameboid | Bandfor | Fimbriat present m ed Sorrated finger | tke edges RBC ‘Stipplin | Stephens | Schiifine | Ziemann | James gs Maurers, | Psdots |‘sdots | dots Gametogony Microgamete | Macrogamete Schizogony 7 [_exeyst A Preenfhvoeyie or | cxooryivocyte eyes _| ‘Gamelo | Banana=sh | Large, | Large, | Large, eye | apedor | round, | round, | round, Number | Fruit pie or Daisy teraz head seat esl 8 i rot te arrange ‘Sporozoite infect liver Py 16 | ment 8 Parenchymal ells ower ——eEe tke | Rosete = ears | form. Schizent | crescent oval cells rupt | Liver cells eupture Sages Ri Stages," Ring form ar releasing the merozoites sametoeyt of, es sehizont 1 sand |B Erythrocytic Cycle trophozo 7 —_ ite) 1 , Merozoies invade RBC ©. Lifecycle r ‘Aside from being vector-borne, it can be transferred ——_1 through contaminated needles, blood transfusion (no. Ring Form ] _ fait stooges verso eel (f) Enytoayc cyl resides inthe RBC 2 E ide the REC or * @) Blooryocgicyde-reeis cutie te RAC a ‘+ Liver cals wil rupture or burst, the schizonts will leave the liver cells and invade the RBC | Rupture of RBC releasing | the merozoites. | [Develop into a macro or macrogamete PAPER PIPER PH |. conv/poeerpipereh ama a RR RAR ARAREAKRAAEARARAARAAAKRAKREAKRBAKBBAABAAASComponents of Malaria Life Cycle Moi er ps ee > rps + Two components: (1) Mosquito vector and (2) Human Host * Prepatent period - parasites are visible '= Incubation period - from the time of infection to clinical onset + Clinical itIness - pathogenicity to recovery Intervals ie Species Prepatent Incubation ‘The malaria parasite fe ycte involves two hosts. During eamoata Farid Period _| a blood meal, a malara-inected female Anopheles a TH-tadays | 8-15daye ‘osqut inocuates sperezetes to the human host fable - Spovazotes infec iver cals and mature into schizo 3 11-15 days | 12-20 days stich ropture and release merozotes «(Of roto, in P —— vivax and P. ovale a dormant stage [hypnezoites} can peo | 3-4weeks 18-40 days Desist inthe ver ana caus relapses by invading the bloodstream weeks, or even years later.) After this initial esmoman, 14-26 days 14-16 days replication in the liver (exo-erythrocytic schizogony 4), the vate parestes undergo asexual mulation in the tnytrooyes (enthrectcschizogory) Merezotes infect Periodicity or Febrile Cycle ‘ed blood calls. The ring stage trophozoites mature into interval | Common Schizons, which yptre releasing merozotes Some (hours)! | Vietins parasites diferentat nto sexual enfrvoeytc sages Fopnie | Paroxysm | (RBCS) (Garetocjes) Blood sage parastes are responsible for Species | Fable | Ttyete {he cnical manifestations ofthe disease. -faggnat | Lifespan: ator every | 120 days ‘The gametocytes, male (microgametocytes) and female a a this hours (macrogametacyes), are ingosad by an Anopheles Plasmodium | Walgnant mosquito during a bload mal. The parasites! falcparum” | "Teran, Tultpeation nthe mosquito fs knoun as te sporogoni suptertan Cycle c- While inthe mosauio’s stomach, the ‘oneal, | 30-48 au mierogametos penetato the macrogameies gonerating Esto. £ygotes «The 2gotesn tum become mote an autumn slongated (ookinetes) 1 which invade the midgut wall of malaria the mosquito where they develop Into oot. The Piasmedum | Benign | ag Youn ooysts grow, rupture, and release sporozoites 12, which vivax. Tertian : | ‘9 make ther way tthe mosqut's salvar glands. Plasmodium | ~Quartan | 75 7a . Inoculation of the sporozates info @ new human host mmalaioe "| mele senescent) Derpetuates the melra if ole Plasmodium | Oval 7 r 4+ Rupltures due to overcrowdedness. ovale Tertian foung 4 Fussion of macragametocytes and microgametocytes: Zygote 'D. Pathogenesis & Clinical Manifestations ‘= Anisocylosis - variation in size ‘= Poiltlocytosis - variation in shape + Sytepasmic vscosty- more schizontsetached inthe BC + RBCstiffness ‘+ Altered REC membrane transport - once the schizont Invade tho RBC, the etacomentof oxygen to REC wi 3e altered PAPERPIPER PH | Acon/oqpermiberh ot See eee VV OK EKYDResistance to infection + Duffy negative Fy(a-b-) —P. vivaxandP knowles) ‘+ Sickle cell anemia Hos = G-6-PD deficiency += Deficiency of paba } P.foleperum PEE Introduction + Fifth species of Plasmodium Pathogenic in primate malarial parasite - Macaca fasciculais : ‘Some records say it can infect human + Similar to Plasmodium Meleriae * PCR assay and molecular characterization MUST-KNOW Black water fever or malarial hemoglobinuria - is a ‘complication of malaria infection in which red blood cells burst in the bloodstream (hemolysis), releasing hemoglobin directly into the blood vessels and into the Urine, frequently leading to kidney failure B. Pathogenesis & Clinical Manifestation Malaria * Recrudescence -reccurencs of parasism wherein ovsr't undergo to exo-erythrocytic oycle ~~ schizonts to invade new RBCs eB falciparum (6-17 months) © _P. malariae (20 years oF more) + Relapse - from dormant to active = from exoerythrocytic eyete, your merozoites inside schizonts wil turn into hyprozotes ~ itil rupture in ver cells, and released > invades RBC © P. vivax (5-7 years) ©. P. ovata (12 months) ‘+ Celebral malaria - CNS © Difluse symmetric encephalopathy, retinal hemorrhages, bruxism, mild neck stiness Classical Malarlal Paroxysyms (Stages of Signs and ‘Sympioms) 1. Cold Stage (© Feeling of intense cold © Vigorous shivering, rigor © _ Lasts 15-60 mins 2. Hot Stage - best stage to collect specimen; reproduction ‘and multiplication 2 Intense heat © Dry burning skin © Throbbing headache © Lasts 2-16 hours 3. Sweating Stage © Profuse sweating © Deciining temperature © Exhausted, weak — si 9 Lasts 2-4 hours ©. Laboratory diagnosis 1. Microscopy > Specimens ‘2. Capillary blood - most preferred b. Venous blood - EDTA treated (has interferences) + Blood flms (Gold standard) ‘rac SMEAR ‘a. Thick flm - large blood volume; for detection (© Should not be fixed (use distlled water to lyse RBC to extract ring form of P. spp.) WBC, platelets & parasites Center - Oil immersion objective Examine at least 100 CF (mahirap makita) Rapid detection and monitoring treatment response oi SMEAR a 'b. Thin film - small blood volume; for identification © Fixed with methanol RBC, WBC, platelets & parasites Feathery edge - Oil immersion objective Examine at least 200-300 CF Species Identification = examine at feathery edge (tongue-shaped ‘wh feathery edge) = darker part: compacted RBC feathery edge: apart RBC. Metarial count: Thick film Giemsa or Wright stain + Light microscopy © WBC and parasites Parasites/ul "200 of WBC tin ‘= += 1-10 parasite! 100 thick field += 11-100 parasite! 100 thick fleld +44 = t-10parasite thick field ++++ = more than 0fthick field 2._Plasmodium antigens diagnostic tests Hi ich protein II- for the carbohydrate release of ttophozoite; detects P. ovale, P. vivax and P. malariae PAPER PIPER PH | th com/agperpiperph e ~—manannnranaen oe 20808790> > > > = = =) = => = => = = > > = > > = = = > > > > > > = > > > > > > > > © Paracheck Pt test 3 ParsHIT fest ‘+ Plasmodium Lactate dehydrogenase - detects P. falsciparum © Diamed Optimat IT + Plasmodium aldolase * usualy for serological examination Antigen detection (test kits) HRP2 assays: | Commercially ParaSight-F | available ‘Simple, rapid falciparum (chloroquine resistant) after patient treated for . vivax (mixed infections) ICT Malaria PF | Commercially | Simple, rapid available lipstick format; | lower false- positive rate; ‘Good sensitivity, especialy when, ‘coupled with microscopy ‘MAKROmed | Commercially | Antigen capture available immuno- chromatographic ‘strip format; more | sensitive than ‘conventional microscopy LOH OptIMAL | Commercially | Simple, rapid available; test | cipstick format positive only with | also picks up P. Viable organisms, | max; few false positives; good test of cure (picks up viable parasites only) 3._QBC Microhematocrit Centrifugation Method = Early detection * Actiine orange staining technique - positive reaction bright green and yellow fluorescent + Fluorescent DNA and RNA stain - using fluorescent ‘microscope 4. Serologic Tests Methods THAT = IFAT » ELISA 5. Molecular Test Methods POR D. Treatment * Chloroquine * Primaquine Therapeutic Use Drugs Aminoquinoione 4, Chloroquine 2. Amodiaquine * Antifolate drugs & combinations ‘Sulfadoxine-pyrimethamine Proguanil CChiorproguanit and dapsone Quinine ‘Astemisinine group Prevention of Malaria Drugs ‘Alovaquone /proguar ‘Chloroquine phosphate Doxycycline Hydroxychioroquine sulate Mefloquine Primaquine - most common © for primary prophylaxis - prevention of acquiring © for terminal prophyiaxis ‘A. Introduction '» Hemosporidian parasite that causes babesiosis, @ hemolytic disease * Tick, splenic, redwater, Texas, or Nantucket fever. + Malaria-lke infections (because ofthe maltese ras, arrangement: because merozoites of malariae's rosette form) ‘+ Vector-bome (hard tick): odes scapularis, © Soft tick: Omithodoros erraticus ‘+ Maltese cross arrangement (Merozoltes & ring form, pear- shaped trophozoite) Infective stage in human: Merozoites Infective stage in vector: Gametes Primary hosts: mammals Intermediate hosts: Ticks B. microt- cattle Morphological forms Intra-erythrocytc forms pleomorphic forms - obscure their dentiication al the species level + Itdoes not: * does not undergo exo-enythrocytic merogony * daughter progeny are not housed in parasitophorous vacuoles + residual bodies are usually non-existent in infected RBCs. Developmental phases 1. merogony in the RBC and in the lick vector 2. stages of gamogony in the gut and epithelium 3. sporogony accompanied with multiple fission in various calls and organs forming sporokinetes, and the development of infective sporozoites PAPER PIPER PH | rcom/poperpiperph 6©. Life Cycle D. Pathogenesis and Clinical Manifestations ‘= Babesia bovis - smaller but more pathogenic '» Babesia bigemina and Babesia cabell-less virulent ‘* Babesia micro - transfer of primed macrophages, as ‘opposed to transfer of primed T-cells, provides protection Babesiosis ‘> Symptoms: one to six weeks post-exposure followed by nnon-periodic or intermittent fever (38 to 40°C), chills, and ‘sweats accompanied with headache, myalgia, arthralgia, nausea, vorniting, and prostration. ‘+ Resolution: mediated by gamma interferon produced by (CD4+ T helper-t cells, alongside macrophage activation ‘Antibodies - useful inthe clearance of extracellular Parasites in circulation. Cerebral babesiosis * caused by infected erythrocytes are sequestered in the * capillary beds © similar manifestation of falciparum malaria mistaken with Lyme disease E. Laboratory Diagnosis 1. Direct microscopic examination of Giemse-stsined peripheral blood smears - definitive diagnosis, acute stage 2. PCR (Gold standard test) Immunofluorescent assays - has drawbacks such as ‘= non-1:16 antibody titer ‘+ Resolution: @ 1:64 serum dilution is highly recommended 4. NODisch-scid mice with the patient's blood - useful in Parasite detection ‘5. Immunochromatographic test (ICT) - diagnosis of both acute and latent infections, ICT strips or dipsticks employed in the detection of infected livestock F. Treatment +” Drug combination of ‘© clindamycin and quinine (drug of choice) ‘clindamycin - 1.2 g intravenously twice a day ‘or 600 mg orally three times a day = quinine ~ 650 mg three times a day ‘© azithromycin and atovaquone - with less adverse effects Both drug combinations are ineffective in suppressing disease progression in immunosuppressed patients * Clindamycin-atovaquone - clear the parasites and prevents recurrence of infection, but produces adverse effects lke Vertigo, tinnitus, and gastrointestinal symptoms * Total blood change - progressive exacerbation of hemolytic anemia © B. equiand B. caballlin vitro - artemisinin, pyrimethamine, ‘and pamaquine ‘© pyrimethamine - effect on dihydrofolate reductase, ‘essential in folate metabolism © pamaquine - interfere in the recycting of endosomal proteins Motaria ss Babosiosis Malaria Babesiosis Matiple rings (except + falsciparum) Large : : trophozoite ‘Gametooyts = Hemozoin . = igment PAPERPIPERPH | :con/openpperph ~~_aeaeaeneanmeaeabeenseneeeaeeeveoeneeaannese eo @ &@ & &