8Entamoeba histolytica (amoebiasis) 2-Precystic form:*Intermediate stage*Low motility*Untrue morphologic stage of parasites*shape: oval or circle
9Entamoeba histolytica (amoebiasis) Precystic formPrecystic form
10Entamoeba histolytica (amoebiasis) 3-Cystic form:*Stable and non motile*Small and large cysts*Primary cyst with -1- nucleus*In replication stage 2-4 nucleus*Karyosome,, chromatoide body and glycogen in nucleus
11Entamoeba histolytica (amoebiasis) 3-Cystic form:Cystic form
12Entamoeba histolytica (amoebiasis) Replication stage of cyst
17Entamoeba histolytica (amoebiasis) Pathogenesis► Cases acute dysentery► Chronic stage: occur intestinal ulcer,inflammation and necrosis with proteolytic enzyme► Heavy infection: occur intestine scleroses ,hypertrophy and perforation► Metastases: to liver and brain…
18Entamoeba histolytica (amoebiasis) Intestinal ameobiasis●Incubation period is 8 days or several month● Acute stage: diarrhea with epithelium butwithout blood, and abdominal pine , lossof weight , flatulence and constipation.● Sever infection : 10-20/day , diarrhea withblood , abdominal pine (colon)dehydration andfever.
19Entamoeba histolytica (amoebiasis) Extra intestinal amoebiais► Spread of intestinal amoeba from blood to liver , spleen,brain and lung.► Direct extra intestinal amoebiasis (without intestinal infection)
20Entamoeba histolytica (amoebiasis) Hepatit Amoebiasis►Causes liver abscess►Single abscess, different size , at the right lob of the liver.
21Entamoeba histolytica (amoebiasis) Hepatic Amoebiasis►Symptoms: liver pine , fever right hypochondrium pain,rarely diarrhea►Transmit to pleura, peritone and pericardial area…..dead.
24Entamoeba histolytica (amoebiasis) Pulmonary Amoebiasis► 1- Direct primary infection (blood circulation)► 2- secondary infection: after liver amoebiasis at the right pulmonary ..Cases are with: single or several abscess
25Entamoeba histolytica (amoebiasis) Cerebral Amoebiasis► Occur from complication ofliver and pulmonary amoebiasis..► Cases are with single or several abscess
26Entamoeba histolytica (amoebiasis Spleen and Cutaneous Amoebiasis► Spleen abscess always seem with liver amoebiasis.► Cutaneous amoebiasis seem in perianal site.
27Entamoeba histolytica (amoebiasis Diagnosis► Detection of trophozoit andcystic form of parasites in fresh stool.► After 30min trophozoit form will destroy.► Extra intestinal amoebiasis:detection of parasite cysts by lugolstain of infected tissue.
28Entamoeba histolytica (amoebiasis trophozoittrophozoitDirect microscopy :Detection of trophozoit and cystic form of parasites in fresh stool.
29Entamoeba histolytica (amoebiasis Treatment► Metronidazole 750mg +diloxanide furoate 500mg X 3 ..10 days..► Metronidazole + Iodoquinol 650mg X days► Metronidazole +tetracycline 250mg X days
30Entamoeba histolytica (amoebiasis OR► Chloroquine 500mg X days (iiver amoebiasis)0R► Paromycine 250mg//kg X days
31Gastrointestinal system protozoon Entamoeba coli
32Entamoeba coli► Entamoeba coli is a non-pathogenic species of Entamoeba that frequently exists as a commensal parasite in the human gastrointestinal tract especially in the colon.
33Entamoeba coli► Clinically, E. coli (not to be confused with the bacterium Escherichia coli) is important in medicine because it can be confused during microscopic examination of stained stool specimens with the pathogenic Entamoeba histolytica
34Entamoeba coli► The presence of E.coli does’nt need treatment treatment as it is considered harmless.► However it should be noted that when a person becomes infected with this benign entamoeba, other pathogenic organisms may have been introduced as well, and these other pathogens might cause infection or illness.
35Entamoeba coli► The identification of intestinal amoebae depends on the size and shape of trophozoites and cysts and on number of nuclei and aspect of karyosome and chromatin.
36Entamoeba coli► Entamoeba coli trophozoites measure µm and have a vescicolous nucleus with a large eccentric karyosome and an irregulary distributed peripheral chromatin. The cytoplasm is vacuolated containing bacteria and yeast.
38Entamoeba coli► E.coli cysts are spherical and measure µm (usually 15-20).► Mature cysts have 8 nuclei with a large karyosome (central or eccentric) and an irregular (sometimes regular) chromatin.► The nuclei can be numerated with careful focusing.►
39Entamoeba coli► While this differentiation is typically done by visual examination of the parasitic cysts via light microscopy, new methods using molecular biology techniques have been developed also.
41Life cycle of E.coliLife cycle of E.coliLife cycle of E.coli
42Entamoeba coli► Cysts and trophosoits of E.coli are larger then E.histolytica.► E.coli is the only species in the genus encountered in humans with more then four nuclei in the cyst stage.
43Entamoeba coli► Cysts and trophosoits of E.coli are larger then E.histolytica.► E.coli is found in the mouth between the gingival pockets.
44Gastrointestinal system protozoon Giardia lamblia (Giardia intestinalis)
45Giardia lamblia ► It is an intestinal flagellate ► Lives in duodenum and jejunum► Cause “Giardiasis”
46Giardia lamblia► Human pathogen: G. lamblia or G. intestinalis or G. duodenalis,► Two life stages: trophozoite and cyst.HISTORY► First observed 1681by Anthony vanLeeuwenhoek
47Giardia lamblia ► First observed 1681 by Anthony van Leeuwenhoek ► Described ~200 years later by Vilem Lambl► First cultured in 1960’s► Confirmed pathogen 1970’s► One of most common intestinal parasites► Causes Giardiasis (beaver fever)►Geographic region with poor water sanitation
48Giardia lamblia ► Species details ► G. intestinalis/lamblia Single-celled protist5 species of Giardia► G. intestinalis/lambliaG. muris in rodents, birds, reptilesG. agilis in amphibiansG. ardae in great blue heronG. psittaci in budgerigar
49Giardia lamblia ► Morphology Cyst ► Infective stage in the environment,► Persist in cold water up toseveral months► Egg-shaped, 8-14µm by 7-10µm► Organelle duplication w/outcytokinesis results*in four nuclei (Nu)*four median bodies (MB)*four axonemes (Ax)
50Giardia lamblia ► Morphology Trophozoite ► Cannot survive in the environment► Motile 4 pairs of flagella► Pear shaped, bilaterallysymmetrical► Relatively flattened, 10-12µm long► 5-7µm wide with a large sucking disk on the anterior ventral side► Two nuclei
51Giardia lamblia - Life cycle - 2 trophozoites released per cystMultiply by binary fission every 12 hSucking disc used for attachmentCyst forms in transit to colon
52Clinical Symptoms and Pathogenesis Giardia lambliaClinical Symptoms and Pathogenesis► Ingested cysts excyst in response to stomach acidity.► Attach (via their sucking disk) to microvilli of epithelium in small intestine, causing epithelial damage and interfering with gut transport processes.
53Clinical Symptoms and Pathogenesis Giardia lambliaClinical Symptoms and Pathogenesis►Epithelial mucus is thinned, lymphocytes and other inflammatory cells infiltrate, physical blocking of absorption may occur, and enterobacteria may proliferate, causing more epithelial damage.►Distention and flatulence can occur.
54Clinical Symptoms and Pathogenesis Giardia lambliaClinical Symptoms and Pathogenesis► Symptoms include profuse and watery to semisolid, greasy, bulky and foul- smelling diarrhea; abdominal cramps; nausea; vomiting; anorexia; low-grade fever and flu-like headache. General malaise, weakness, weight loss.
55Giardia lamblia► Symptoms more likely in children and immunosuppressed adults than in healthy adults.► Infectous dose is low (about 10 cysts; possibly less).► Incubation period from 7-11 days.
56Giardia lamblia ► Duration of illness varies: few days to months. ►Infection may resolve spontaneously. often, subchronic stage develops with mild to moderate symptoms occurring periodically.
57Giardia lambliaEpidemiology► Many animals are major reservoirs: dogs, mice, horses, sheep….► Contamination is by “Fecal-oral route”► Transmission by contaminated water and food is common► Transmission is generally in day-care centers, with poor hygiene and sanitation.
58Giardia lambliaEpidemiology► Levels in sewage are estimated in the range of 3,000 to 30,000 per liter.► Animal-to-human transmission occurs, but pathogenicity and virulence of non-human sources for humans is still uncertain.► Strain differences documented both antigenically and genetically.
59Giardia lamblia.“Diagnosis” ► Diagnosis is by microbiological examination of stool samples to check for the parasite.► Because Giardia can be difficult to diagnose, the patient will ask to submit multiple stool specimens collected over a few days.
60intestinalis trophozoite in a wet mount stained with iodine. G.lamblia cysts stained by metilen blueintestinalis trophozoite in a wet mount stained with iodine.intestinalis trophozoite in a wet mount stained with iodine.G.lamblia trophozoite in a wet mount stained with iodine.60
61Giardia lamblia :Diagnosis ► Microscopic detection based on finding distinctive trophs. or cysts in stools, with our without dye staining or immunofluorescent (fluorescent antibody) staining.► Newer clinical diagnostic detectionby enzyme immunoassay (EIA; ELISA).►Molecular detection by PCR is nowbeing used for detection and characterization.
62Giardia lamblia Visualization methods ► Laser scanning cytometry► Immunofluorescent antibodies(Giardia –specific) for enumeration► Differential interference contrast(DIC) for confirmation4,6-diamidino 2phenyl-indole dihydrochloride (DAPI)for viability► Propidium iodide (PI) for non-viability
64Environmental Control of Giardia ► Water: physical and chemical treatment(coagulation-flocculation, sedimentation, filtration and disinfection) will reduce Giardia►Relatively resistant to chemical desinfectionbut relatively sensitive to physical disinfection by UV radiation or heat.► Pasteurization and thermal treatments iseffective for foods
65Giardia lamblia ► Washing hands thoroughly with soap and water. Protection from Giardiasis► Washing hands thoroughly with soap and water.► Not drinking untreated water or not use ice made from untreated water .► Washing all food that is to be eaten raw by uncontaminated water..
66Giardia lamblia Treatment ► Quinacrine HCl ,Metronidazole (Flagyl) and Furazolidone (Furoxone).► Oral Quinicrine HCl cures about 90% of infections.► Probiotics could help thetreatment
67Giardia lamblia ► Fluid replacement is an ► Giardia can infect all people ,youngchildren and pregnant women .► Rapid loss of fluids fromdiarrhea can be especiallylife threatening .► Fluid replacement is animportant part of the treatment