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REVISION Statements True OR False The number of nucleus found in mature cyst of E. histolytica is 8. Ingested RBC usually found in the E. histolytica trophozoites. Asymptomatic individuals with documented E. histolytica infection should be no...

REVISION Statements True OR False The number of nucleus found in mature cyst of E. histolytica is 8. Ingested RBC usually found in the E. histolytica trophozoites. Asymptomatic individuals with documented E. histolytica infection should be not treated. The life cycle of E. coli is same as E. histolytica except that it remains a luminal commensal without tissue invasion. A chromatoid body is a clump of genetic material found in the nucleus of amoebae. REVISION 1. The parasitic form called ______ is sensitive to environmental changes and it disintegrates rapidly upon release from the body. 2. The morphological forms of Naegleria fowleri can be _____, ______ and ______. 3. Peripheral nuclear chromatin is absent in ______ and ______. 4. Free living amoeba include ______ and ______. 5. Diseases caused by Acanthamoeba spp. are ______ and ______.  CSI153 Clinical Parasitology Protozoa: Intestinal and atrial flagellates and ciliates Nurhidayah binti Ab. Rahim Mac 2019 Diploma Teknologi Makmal Perubatan, UiTM (Pulau Pinang), Kampus Bertam  Learning outcomes Students should be able to: Features/ Life cycle characteristic Infection and Mode of infection diagnostic stage Diagnosis and treatment Objective 2: Classification of protozoa PROTOZOA of MEDICAL IMPORTANCE phyla Sarcomastigophora Ciliophora Apicomplexa Microspora In colon: Microsporidia subphyla Balantidium coli Subphylum Sarcodina (= Amoeba) In colon, liver and lungs: In intestine: 1. Entamoeba histolytica 1. Cryptosporidium parvum, 2. Cyclospora cayetanensis, Subphylum Mastigophora (= Flagellates) 3. Sarcocystis species In small intestine: In blood: 1. Giardia lamblia, 4. Plasmodium species On mucous membranes of vagina: In tissue: 1. Trichomonas vaginalis 5. Toxoplasma gondii 5  Protozoa Intestinal Amoeba Free living Amoeba Intestinal and atrial Entamoeba histolytica flagellates and ciliates Entamoeba coli Naegleria fowleri Endolimax nana Giardia lamblia Acanthamoeba spp. Iodamoeba butschlii Dientamoeba fragilis Blastocystis hominis Trichomonas vaginalis Balantidium coli Blood and tissue sporozoa Blood and tissue flagellates Plasmodium vivax Plasmodium ovale Leishmania spp. Plasmodium malariae Trypanosoma spp. Plasmodium falciparum Plasmodium knowlesi Babesiosis Toxoplasma gondii Pneumocystis carinii Intestinal and atrial flagellates and ciliates Giardia lamblia Giardia lamblia Distribution  Worldwide, particularly in England, Russia, several countries in Eastern Europe and many seaside areas of the Mediterranean  Found in lakes, streams and other water sources Kingdom: Protista Subkingdom: Protozoa Phylum: Sarcomastigophora Subphylum: Mastigophora Intestinal Class: Zommastigophora flagellate Order: Diplomonadida Family: Hexamitidae Genus: Giardia Species: lamblia Transmission  Fecal-oral route  Ingestion of food and water contaminated with animal or human feces Intestinal and atrial flagellates and ciliates Giardia lamblia  Pear shape and 8 flagella  Length: 9-21 mm  Width: 5-15 mm  Bilaterally symmetrical  2 nucleus  One on either side of axostyle  Fine granular cytoplasm  2 median parabasal body Lateral view (look like mustache) Bilateral: having two sides Intestinal and atrial flagellates and ciliates Giardia lamblia Left: Kohn stain. Center: Trichrome. Right: in vitro culture, from a quality control slide. Intestinal and atrial flagellates and ciliates Giardia lamblia  Oval and lightly bile-stained  Diameter 8-12 mm  4 nucleus (mature)  Small eccentric karyosome  Clear space beneath the thin cyst wall Intestinal and atrial flagellates and ciliates Giardia lamblia excystation encystation Intestinal and atrial flagellates and ciliates Giardia lamblia  Infection occurs by the ingestion of cysts in contaminated water, food, or by the fecal-oral route.  In the small intestine, excystation releases trophozoites.  Trophozoites multiply by longitudinal binary fission, remaining in the lumen of the proximal small bowel where they can be free or attached to the mucosa. Intestinal and atrial flagellates and ciliates Giardia lamblia Life cycle of Giardia lamblia Encystation (trophozoites multiply) occurs as the parasites transit towards the colon. The cyst is the stage found most commonly in nondiarrheal faeces Because the cysts are infectious when passed in the stool or shortly afterward, person-to- person transmission is possible. Intestinal and atrial flagellates and ciliates Giardia lamblia Pathogenesis  Giardia lamblia causes giardiasis (inflammation due to Giardia)  It inhabit glandular crypts of mucosa layer in duodenum and jejunum  It attach to epithelial surface of mucosa layer  Sometimes disrupt intestinal function  malabsorption  weight loss and Attach to epithelial surface of mucosa steatorrhea (excessive amount of fat layer in the faeces)  Malabsorption of fat  acute diarrhea and abdominal pain ( high fat content in stool)  Some patients asymptomatic  Some patients have general symptoms  Fever  Anaemia  Allergy Intestinal and atrial flagellates and ciliates Giardia lamblia Laboratory diagnosis Stool Duodenal Serum fluid - detect antibody against Giardia Macroscopic Microscopic Serology test - No blood and mucus examination - Not useful in diagnosis as presence antibody might be present due to past infection Microscopic examination - Direct wet mount (saline ELISA and iodine) Detection of motile Indirect immunofluorescent trophozoites in liquid stool - Using antibody to detect Graceful falling-leaf Giardia antigen motility  Detect Giardia which ELISA:An enzyme is used when the enzyme and the missed out under temporary antigen-antibody complex Permanent bind, astaining and permanent color indicator or other easily recognizable stain (high sign occurs. - Kohn staining sensitivity) - Trichrome staining Graceful falling-leaf motility Intestinal and atrial flagellates and ciliates Giardia lamblia Stool specimen Examination of triple fecal test – examination of 3 stool specimens from nonconsecutive day Due to trophozoite attach to mucose membrane firmly, sometimes fecal specimen has no trophozoite  Concentration technique and permanent stain smear Detection of cyst in formed stool Intestinal and atrial flagellates and ciliates Giardia lamblia Intestinal and atrial flagellates and ciliates Giardia lamblia *Kohn stain Intestinal and atrial flagellates and ciliates Giardia lamblia  Treatment  Quinacrine or metronidazole  Furazolidone for children  Prevention  Avoiding contaminated food and water  Proper disposal of feces  Good personal hygiene  Proper storage of food and water  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  Genital flagellates  Found in vagina and urethra  All Trichomonas species have  3-5 anterior flagella  One undulating membrane Kingdom: Protista  Distribution Subkingdom: Protozoa Phylum: Sarcomastigophora  Common worldwide Subphylum: Mastigophora  Transmission Family: Trichomonadidae  Sexual contact ( higher prevalence among persons with multiple sexual partners)  Contaminated towels  Contaminated examination equipment  Infants may be infected during birth  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  Pear shape, round anterior, pointy posterior  Length: 10-30 mm  Width: 5-20 mm  Ovoid nucleus at anterior  4 anterior flagella  Undulating membrane from anterior to middle of body  Axostyle from anterior to posterior and end like a tail  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis *Giemsa stain  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  Transmitted by sexual contact  Inhabit in vaginal of females and urethra of males  Only exist in trophozoite form, does not appear to have cyst form  Multiple by longitudinal binary fission  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  Trichomonas vaginalis is the main parasite which causes trichomoniasis  In males  Infect urethra, seminal vesicle and prostate  Usually asymptomatic  10% have urethritis which produce thin, white urethral discharge  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  In females  Infect vulva, vaginal and cervix, usually not extend to uterus  Cause vaginitis which produce offensive yellowish discharge  Trichomonas vaginalis grow optimally at pH 5.5-6.0  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis a) Microscopic examination i) Detection of motile trophozoite in a wet preparation of vaginal or urethral discharge. - Discharge + one drop of saline, put on glass slide, + cover slip - Trophozoite has jerky movement (very sudden and quick, and do not flow smoothly.) ii) Detection of trophozoite in Papanicolau smear ( Pap smear)from the cervix (common used and for females only) - Common stain used is Papanicolau, Leishman and Giemsa - Immunoflourescent stain iii) Detection of trophozoite in urine or prostatic secretions in males b) Culture - Trichomonas vaginalis in vaginal and urethral discharge can be isolated using culture media - Recommended when direct smear microscopy is negative  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis c) Molecular method  DNA hybridisation method  Polymerase chain reaction (PCR)  Intestinal and atrial flagellates and ciliates Trichomonas vaginalis  Treatment  Metronidazole – topical and systemic  Sexual partner must also be treated to prevent recurrence of infection  Prevention  Detection and treatment of cases  Avoid unprotected sexual contact  No vaccine available at present  This patient was suffered with critical weight lost and steatorrhea. She had fever, anaemia and allergy. Name the parasite that cause the infection and draw it life cycle. Intestinal and atrial flagellates and ciliates Dientamoeba fragilis Dientamoeba fragilis Morphology Binucleate form Spindle structure between the two nuclei Site of infection Gastrointestinal tract Route Oral-fecal transmission Symptoms Diarrhea, abdominal pain, flatulence*, nausea, fatigue Disease Dientamoebiasis *the accumulation of gas in the alimentary canal. Intestinal and atrial flagellates and ciliates Dientamoeba fragilis C C Parasitology for Medical and Clinical Laboratory Professionals, John Ridley, 2011, Cengage Learning MICROCOPIC DIAGNOSTIC FEATURES Parasitology for Medical and Clinical Laboratory Professionals, John Ridley, 2011, Cengage Learning Intestinal and atrial flagellates and ciliates Dientomoeba fragilis *Trichome staining  Intestinal and atrial flagellates and ciliates Balantidium coli  Phylum: Ciliophora  Family: Balantididae  Only ciliate protozoan parasite of human and largest protozoan  Distribution: World-wide  Natural host: pigs  Accidental host: man  Reservoirs: monkeys, pigs, rats  Site: large intestine  Intestinal and atrial flagellates and ciliates Balantidium coli Trophozoite Cyst (infective stage) Active motile – short delicate cilia Motile over the entire surface of body Large ovoid cells spherical 2 nucleus 2 nucleus -Large kidney-shaped macronucleus -macronucleus -Small micronucleus -micronucleus Cytoplasm Cytoplasm -1-2 contractile vacuoles -vacuoles -several food vacuoles Cilia: present Cilia: present - Surrounded by a thick and transparent double-layered wall  Intestinal and atrial flagellates and ciliates Balantidium coli trophozoite cyst *Trichrome stain  Intestinal and atrial flagellates and ciliates Balantidium coli Pathogenesis -Mucosal ulcers and submucosal abcesses -Do not invade liver or any extraintestinal sites Diagnosis - stool examination - Biopsy (intestinal ulcers) - Cultures (locke’s egg albumin medium or NIH polyxenic medium Treatment - tetracycline - metronidazole -Transverse binary fission (asexual) - nitroimidazote -Conjugation (sexual) Draw life cycle of: Giardia Trichomonas Dientomoeba Balantidium lamblia vaginalis fragilis coli  Giardia Trichomonas Dientomoeba Balantidium lamblia vaginalis fragilis coli Trophozoites cyst  CSI153 Clinical Parasitology Protozoa: Blood and tissue flagellates Nurhidayah binti Ab. Rahim April 2018 Diploma Teknologi Makmal Perubatan, UiTM (Pulau Pinang), Kampus Bertam  Protozoa Intestinal Amoeba Free living Intestinal and atrial Entamoeba histolytica Amoeba flagellates and ciliates Entamoeba coli Naegleria fowleri Endolimax nana Giardia lamblia Acanthamoeba spp. Iodamoeba butschlii Dientamoeba fragilis Blastocystis hominis Trichomonas vaginalis Balantidium coli Blood and tissue sporozoa Blood and tissue flagellates Plasmodium vivax Plasmodium ovale Leishmania spp. Plasmodium malariae Trypanosoma spp. Plasmodium falciparum Plasmodium knowlesi Babesiosis Toxoplasma gondii Pneumocystis carinii  Blood and tissue flagellates Leishmania -Obligate intracellular parasite -2 hosts: mammalian, insect vector (female sandfly) - site: macrophages (human and other mammalian)–---- - Mode of transmisson: blood-sucking sandflies amastigote form promastigote form (intracellular) (extracellular) ovoid body (nucleus, spindle shape body, single kinetoplast, non-motile, no flagellum (anterior part), flagellum, longitudinal binary motile, longitudinal binary fission) fission Human Female sandfly Blood and tissue flagellates  Blood and tissue flagellates Promastigote Amastigote Blood and tissue flagellates  Blood and tissue flagellates Pathogenesis - Cutaneous leishmaniasis - Mucocutaneous leishmaniasis - Visceral leishmaniasis (liver, spleen) Cutaneous leishmaniasis Diagnosis - Biopsy (intestinal ulcers) - Cultures (nutrient agar+blood) - serological and molecular test Treatment - pentavalent antimonials (sodium stibogluconate or meglumine antimonate) Mucocutaneous leishmaniasis Visceral leishmaniasis  Trypanosoma spp. Self reading - Morphology - Mode of transmission - Life cycle - Pathogenesis - Diagnosis - Treatment 24 APRIL 2018, 10 – 12 PM, MAKMAL KOMPUTER 1 & 2