PROTOZOA FLAGELLATES

These are Protozoa using flagella for locomotion and food procurement. 1. Taxonomical classification Subphylum Sarcomastigophora SuperclassMastigophora- Moving by flagella Class Zoomastigophora Lacking chromatophores nutrient holozoic, reproduction asexual. 2. Divided into two groups: A. Those living in digestive tract and genitalia, transmission from person to person without biological vector. I.e. Giardia lamblia, Chilomastix mesnili, Trichomonas Spp of genitalia and GI tract

B. Haemoflagellates- Parasites of blood stream and tissue, requiring blood sucking arthropods as biological vectors Family Trypanosomatidae 1. Genus Leishmania- Spp for human Leishmaniasis 2. Genus Trypanosoma -Spp-for Leishmania humans Trypanosomiasis

Trypanosoma

GIARDIA LAMBLIA

Note ventral adhesive discs

Giardia lamblia Trophozoites

Giardia lamblia (Intestinalis)

Causes Human giardiasis Phylum (Protozoa) Class: ZOOMASTIGOPHOREA

Order:
Family

Diplomonadidae
Hexamitidae--- (Two nuclei lying side by side)

Genus GIARDIA Species Giardia lamblia (Intestinalis)

Morphology & Life cycle of Giardia lamblia1

Giardia lamblia It colonises and reproduces in the small intestine, causing Giardiasis. Giardia lamblia has both cyst and trophozoite forms. Cyst has four nuclei and it is the Infective stages of man Trophozoite is the vegetative and reproductive and pathogenic stage . It has 8 flagella and two nuclei Both cysts and trophozoites can be found in lose stool

Major characteristics
Distinguishing features of the trophozoites Two nuclei Large karyosome and no peripheral chromatin, giving the two nuclei a halo appearance. Cysts are distinguished by a retracted cytoplasm. The protozoan lacks mitochondria,

Causes Human giardiasis Under a normal compound light microscope, Giardia often looks like a "clown face," with two nuclei outlined by adhesive discs above dark region that forms the "mouth." Cysts are oval, have four nuclei, and have clearly visible axonemes Note the 2 anterior nuclei (eyes), (exostyle). the ventral adhesive disc, the Giardia lamblia has no axostyle and the 8 flagella. mitochondria

Giardia trophozoite

Life cycle

Life cycle

Life cycle of Giardia lamblia2

Transmission: Infection is acquired by ingestion of mature cyst- the resistant forms found in the environment Excystation: Cysts undergo excystation in small intestine to releases trophozoites Cysts pass through the stomach, exposed to gastric acid/low pH and emerge in the lower stomach or upper small intestine as a trophozoite. Each cyst has four nuclei and produces 2 trophozoites

Life cycle of Giardia lamblia3 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 by a ventral sucking disk Encystation: Trophozoites migrating to the distal small intestine undergo a process of encystation as moisture in faeces dwindles , and pH becomes acidic with increased effect of bile salts

Life cycle of Giardia lamblia4

Encystation: Encystation requires the synthesis of the cell wall that will resist the hash environment outside
In encystation trophozoite retract the flagella into axonemes and assume the appearance of 4 pairs of curve bristles Cytoplasm becomes thin and a tough hyaline membrane is secreted to form the cyst wall. Cysts: They have rigid cell walls and are non-dividing, can survive several months in cold water. Cysts can resist desiccation in the environment

Life cycle

 Epidemiology: Giardia infection can occur through ingestion of dormant cysts in contaminated water, food, or by the fecal-oral route (through poor hygiene practices). Cyst can survive for weeks to months in cold water, found in contaminated wells and water systems, and even clean-looking streams. May occur in city reservoirs and persist after water treatment, Giardia cysts are resistant to conventional water treatment methods such as chlorination Boiling water is recommended

Transmission Faecal-oral transmission may occur, in day care centres due to poor hygiene practices. Children workers, family members of infected individuals are at risk of infections Not all Giardia infections are symptomatic, many people serve as carriers of the parasite.

Transmission2 Risks of infection are increased by reservoir hostsGiardia is the Most common parasites infecting cats, dogs Cows, sheep, deer (in recreational areasZoonotic transmission: Person-to-person transmission: Possible when cysts are passed in the stool or shortly afterward Pathogenesis, symptoms and signs Characteristic location is of the trophozoites is the glandular crypts of duodenal-jejunal mucosa. Do not invade the tissue but feed on mucosa secretions, may damage villi

Pathogenesis2: Incubation period 5-20 days Large surface of mucosa covered by trophozoites interfering with fat absorption hence steatorrheic type of stool with excess secretion mucus and fat. Malabsorption may cause vitamin B12 deficiency in chronic infection People with recurring Giardia infections, particularly those with a lack of IgA, may develop chronic disease. Lactase deficiency may develop Gallbladder may be parasitized causing blockage of bile duct- may result in jaundice Duodenal irritation dehydration and dull epigastric pain may occur

Symptoms2
It usually causes "explosive diarrhoea" and while unpleasant, is not fatal. In healthy individuals, the condition is usually self-limiting, although the infection can be prolonged in patients who are immunocompromised, or who have decreased gastric acid secretion.

Symptoms of infection include (in order of frequency) diarrhoea, malaise, excessive gas. (often flatulence or a foul or sulphuric-tasting belch, known to be so nauseating in taste that it can cause the infected person to vomit), Clinical manifestations- It is a clinical disease of mainly infants and children Clinical Disease is Self limiting in 4-7 weeks There is Intermittent shedding of cysts Acute diarrhoea- 5 -7days, abdominal cramps & bloating, Anorexia & feverdiminished interest in food, possible vomiting and weight loss. Pus and blood are not commonly present in the stool.

Diagnosis
Diagnosis of Giardiasis is done by observation of cysts and/or trophozoites in faeces or duodenal aspirates under a microscope Multiple microscope stool examinations are recommended, since the cysts and trophozoites are not shed consistently. Accurate diagnosis may require an antigen test especially in none endemic areas

Giardia lamblia trophozoites & cysts

cysts

Treatment:
Human infection is conventionally treated with metronidazole, tinidazole or nitazoxanide. Given the difficult nature of testing to find the infection, including many false negatives, some patients should be treated on the basis of empirical evidence;

Prevention
Chlorine treatment of drinking water for Giardia is not very successful in killing the organism. Reliable prevention of outdoor water typically involves filtration with a filter that has a nominal 1-micrometer pore size. Most chemical treatment methods, including common point-of-use treatments such as iodine and chlorine dioxide, are considered unreliable in inactivating Giardia cysts. Water parameters such as temperature, turbidity, and dissolved solids may also affect the effectiveness of such treatments.

Cyst of G. lamblia
THANK YOU

& HAVE A GIARDIA FREE DAY Chilomastix mesnili

Other flagellates
Chilomastix mesnili

Chilomastix mesnili
Chilomastix mesnili -is a harmless commensal flagellate whose presence in a stool specimen suggest hand to mouth faecal contamination It has well defined trophozoite and cystic stages Trophozoite stage has a spiral groove extending through the middle half of the body giving it a pear shape The size is about 6 20 microns in length and 3 10 microns in breath. It has a nucleus measuring 3 4 microns medially situated near the pole with distinct central karyosome

Cyst of Chilomastix mesnili

Cysts are pear-lemon shaped, colourless measuring 7 -10 microns in length x4 6 microns in breadth Chilomatix mesnili is a normal inhabitant of coecum where the trophosites live on enteric bacteria in the lumen of glands and multiply by binaly fission

CILIATES

CILIATES- Balantidium coli

The ciliates belong to the family Ciliophora. They possess simple cilia or compound ciliary organelles This are extensions of ectoplasmic membranes and are used for locomotion and attachments. They have 2 types of nuclei and a large contractile vacuole. They have cystostome which they use as cell mouth and Cytopyge as anal pore Balantidium coli is the only representative protozoa in man Balantidium Spp have been reported as natural parasites in hogs, monkies, cockroaches etc.

Balantidium coli
Balantidium coli is the only member of the ciliate family to cause human disease It is the largest protozoa that parasitize human The organism is widely distributed in warmer climates where human infections most commonly occur. It inhabit the large intestine, caecum and terminal ileum where they feed on bacteria There are both cyst and trophozoite stages

Balantidium coli
Classification Phylum Protozoa Class Ciliata The disease is known as balantidiosis, or balantidial dysentery Transmitted by fecaloral route

Morphology: Trophozoites Trophozoites of B. coli measures 50-150m in length and 40-70 mm in width but have been known to attain lengths of up to 200m They are oval in shape and covered in short cilia. The cilia are present in young cysts and may be seen slowly rotating, But after prolonged encystment, the cilia disappear. A funnel shaped cytosome (cell mouth) can be seen near the anterior end and a cytophage at the other end. The cytoplasm contains numerous food vacuoles and one or two contractile vacuoles. Nuclei: It has two nuclei- One large bean shaped macronucleus, micronucleus in the centre of the curvature of macronucleus.

Life cycle
Reproduction by Traverse binary fission Cytoplasm divide into two daughter cells Nuclei do not multiply when encysted

Clinical Disease
Severe B. coli infections may resemble amoebiasis. Symptoms include diarrhoea, nausea, vomiting and anorexia. The diarrhoea may persist for long periods of time resulting in acute fluid loss. B. coli also has the potential to penetrate the mucosa resulting in ulceration. Extra-intestinal disease has also been reported but rarely.

Laboratory Diagnosis
Wet preparations of fresh and concentrated stool preparations reveal the characteristic cysts and trophozoites. They are easier to identify in wet preparations than permanently stained faecal smears. In unstained preparation, the organisms are easily recognized because of their size and rapid revolving rotation. In a stained preparation, the characteristic macro and micronuclei may be observed

 Balatidium coli -Commensal Note macronuclei A) A cyst of B. coli, the only ciliate parasite of humans, in an unstained wet mount of formalin-preserved faeces. The large macronucleus is visible as a clear area at the right side of the cyst. B) A trichrome-stained trophozoite of B. coli. The cytostome is seen at the top of the organism and the macronucleus is the dark-staining structure located in midbody. Cilia are visible on the surface as hair-like projections. May cause diarrhoea/dysentery

Note macronuclei A B

THANK YOU

GIARDIASIS Manifestation of infection Colonization of the gut results in inflammation and villous atrophy, Reduced the gut's absorptive capability. In humans, infection is symptomatic only about 50% of the time,.

 Symptoms It usually causes "explosive diarrhea" and while unpleasant, is not fatal. In healthy individuals, the condition is usually selflimiting, although the infection can be prolonged in patients who are immunocompromized, or who have decreased gastric acid secretion. People with recurring Giardia infections, particularly those with a lack of IgA, may develop chronic disease. Lactase deficiency may develop in an infection with Giardia, however this usually does not persist for more than a few weeks before a full recovery

Symptoms
Symptoms of infection include (in order of frequency) diarrhea, malaise, excessive gas (often flatulence or a foul or sulphuric-tasting belch, which has been known to be so nauseating in taste that it can cause the infected person to vomit), steatorrhoea (pale, foul smelling, greasy stools), epigastric pain, bloating, nausea, diminished interest in food, possible (but rare) vomiting which is often violent, weight loss. Pus and blood are not commonly present in the stool.

 Giardiasis cause of Vitamin B12 deficiency, due to intestinal malabsorption of fats.

Diagnosis
Tratment: Given the difficult nature of testing to find the infection, including many false negatives, some patients should be treated on the basis of empirical evidence; (treating based on symptoms). Human infection is conventionally treated with metronidazole, tinidazole or nitazoxanide. Although Metronidazole is the current first-line therapy, it is mutagenic in bacteria and carcinogenic in mice, so should be avoided during pregnancy.[3] One of the most common alternative treatments is (found in Oregon grape root, goldenseal, yellowroot, and various other plants).[citation needed] Berberine has been shown to have an antimicrobial and an antipyretic effect.[citation needed] Berberine compounds cause uterine stimulation, and so should be avoided in pregnancy.[citation needed] High doses of berberine can cause bradycardia and hypotension. [11]

 Microscopy Under a normal compound light microscope, Giardia often looks like a "clown face," with two nuclei outlined by adhesive discs above dark that form the "mouth." Cysts are oval, have four nuclei, and have clearly visible axostyles. In spite of the common belief that all Eukaryotes have mitochondria, Giardia is one of the few that lack these organelles

DrugTreatment Side Effects

Metronidazole5-7 daysMetallic taste; nausea; vomiting; dizziness; headache; disulfiram-like effect; neutropeniaTinidazole Single doseMetallic taste; nausea; vomiting; belching; dizziness; headache; disulfiram-like effect Nitazoxanide 3 daysAbdominal pain; diarrhea; vomiting; headache; yellow-green discolouration of urineTable adapted from Huang, White

Dientamoeba fragilis
Trophozoites of Dientamoeba fragilis has two nuclei.. The karyosomes within the nuclei are variable in appearance; often they are fragmented into 3 to 8 pieces but in other instances they may appear as a single mass. There is no peripheral chromatin on

Dientamoeba fragilis- Trophozoites

Other flagellates
Chilomastix mesnili

Chilomastix mesnili
Chilomastix mesnili -is a harmless commensal flagellate

whose presence in a stool specimen suggest hand to mouth faecal contamination hence possibility of amoeba infection. It has well defined trophozoite and cystic stages Trophozoite stage has a spiral groove extending through the middle half of the body giving it a pear shape The size is about 6 20 microns in length and 3 10 microns in breath. It has a nucleus measuring 3 4 microns medially situated near the pole with distinct central karyosome

Cyst of Chilomastix mesnili

Cysts are pear-lemon shaped, colourless measuring 7 -10 microns in length x4 6 microns in breadth Chilomatix mesnili is a normal inhabitant of coecum where the trophosites live on enteric bacteria in the lumen of glands and multiply by binaly fission

Chilomastix mesnili (Flagelate)

CILIATES
The ciliates belong to the family Ciliophora. They possess simple cilia or compound ciliary organelles This are extensions of ectoplasmic membranes and are used for locomotion and attachments. They have 2 types of nuclei and a large contractile vacuole. They have cystostome which they use as cell mouth and Cytopyge as anal pore Balantidium Spp have been reported as natural parasites in hogs, monkies, cockroaches etc.

Balantidium coli
Balantidium coli is the only member of the ciliate family to cause human disease It is the largest protozoa that parasitize human The organism is widely distributed in warmer climates where human infections most commonly occur. The organisms inhabit the large intestine, caecum and terminal ileum where they feed on bacteria There are both cyst and trophozoite stages

Balantidium coli
Classification Phylum Protozoa Class Ciliata It is the only ciliated protozoa infectious infectious for humans The disease is known as balantidiosis, balantidiosis, or balantidial dysentery Transmitted by fecal-oral route

Morphology: The cyst is spherical or ellipsoid and measures from 50 - 70m. It contains 1 macro (large bean shaped)and 1 micronucleus (within macronucleus). The cilia are present in young cysts and may be seen slowly rotating, But after prolonged encystment, the cilia disappear.

Morphology
Trophozoites Trophozoites of B. coli measures 50-100m in length and 40-70 mm in width but have been known to attain lengths of up to 200m They are oval in shape and covered in short cilia. A funnel shaped cytosome (cell mouth) can be seen near the anterior end and a cytophage at the other end. The cytoplasm contains numerous food vacuoles and one or two contractile vacuoles.
It contains 1 macronucleus (large bean shaped) and 1 micronucleus (in the centre of the curvature of macronucleus).

In an unstained preparation, the organisms are easily recognized because of their size and rapid revolving rotation. In a stained preparation, the characteristic macro and micronuclei may be observed.

Life cycle

Clinical Disease
Severe B. coli infections may resemble amoebiasis. Symptoms include diarrhoea, nausea, vomiting and anorexia. The diarrhoea may persist for long periods of time resulting in acute fluid loss. B. coli also has the potential to penetrate the mucosa resulting in ulceration. Extra-intestinal disease has also been reported but rarely.

Laboratory Diagnosis
Wet preparations of fresh and concentrated stool preparations reveal the characteristic cysts and trophozoites. They are easier to identify in wet preparations than permanently stained faecal smears.

 THANK YOU

Giardia Life cycle

The life cycle begins with a non-infective cyst being excreted with the faeces of an infected individual. Distinguishing characteristic of the cyst is four nuclei and a retracted cytoplasm (infective stage through ingestion). Once ingested by a host, the trophozoite emerges to an active state of feeding and motility. Trophozoite undergoes asexual replication through longitudinal binary fission. The resulting trophozoites and cysts then pass through the digestive system in the faeces. While the trophozoites may be found in the faeces, only the cysts are capable of surviving outside of the host.

Symptoms2
It usually causes "explosive diarrhea" and while unpleasant, is not fatal. In healthy individuals, the condition is usually self-limiting, although the infection can be prolonged in patients who are immunocompromised, or who have decreased gastric acid secretion.[3] People with recurring Giardia infections, particularly those with a lack of IgA, may develop chronic disease. Lactase deficiency may develop in an infection with Giardia infection,

Diagnosis
Giardia lamblia infection in humans is frequently misdiagnosed. Accurate diagnosis requires an antigen test or, if that is unavailable, an ova and parasite examination of stool. Multiple stool examinations are recommended, since the cysts and trophozoites are not shed consistently.

Tratment: Given the difficult nature of testing to find the infection, including many false negatives, some patients should be treated on the basis of empirical evidence; (treating based on symptoms). Human infection is conventionally treated with metronidazole, tinidazole or nitazoxanide. Although Metronidazole is the current first-line therapy, it is mutagenic in bacteria and carcinogenic in mice, so should be avoided during pregnancy.[3] One of the most common alternative treatments is (found in Oregon grape root, goldenseal, yellowroot, and various other plants).[citation needed] Berberine has been shown to have an antimicrobial and an antipyretic effect.[citation needed] Berberine compounds cause uterine stimulation, and so should be avoided in pregnancy.[citation needed] High doses of berberine can cause bradycardia and hypotension. [11]

Giardia lamblia trophozoites & cysts

 As well as waterborne sources, fecal-oral transmission can also occur, for example in day care centres, where children may have poor hygiene practices. Those who work with children are also at risk of being infected, as are family members of infected individuals. Not all Giardia infections are symptomatic, and many people can unknowingly serve as carriers of the parasite.

Treatment in animals Cats can be cured easily, lambs usually simply lose weight, but in calves the parasites can be fatal and often are not responsive to antibiotics or electrolytes. Carriers among calves can also be asymptomatic. Dogs have a high infection rate, as 30% of the population under one year old are known to be infected in kennels. The infection is more prevalent in puppies than in adult dogs. This parasite is deadly for chinchillas, so extra care must be taken by providing them with safe water. Infected dogs can be isolated and treated, or the entire pack at a kennel can be treated together regardless. Kennels should also be then cleaned with bleach or other cleaning disinfectants. The grass areas used for exercise should be considered contaminated for at least one month after dogs show signs of infection, as cysts can survive in the environment for long periods of time. Prevention can be achieved by quarantine of infected dogs for at least 20 days and careful management and maintenance of a clean water supply.

Giardia lamblia (Lamblia intestinalis,

Giardia duodenalis) Life cycle
Giardia lamblia is a flagellated protozoan It colonises and reproduces in the small intestine, causing Giardiasis. It attaches to the epithelium by a ventral adhesive disc, Reproduction is by longitudinal binary fission. Giardiasis remains confined to the lumen of the small intestine, does not spread via the bloodstream, nor does it spread to other parts of the gastro-intestinal tract, Giardia trophozoites absorb their nutrients from the lumen of the small intestine, and are anaerobes. If the organism is split and stained, it has a very characteristic pattern that resembles a smiley face.

Giardia
Giardia has trophozoite and cyst stages Trophozoite stage is pear shaped (10-20m long, Rounded anteriorly and pointed posterioly) Has two nuclei with central karyosome and single dense chromatine mass and: 8 flagellum (Four on lateral surface Two vertical and two posteriorly (tail) Sucking disk (Bow shaped depression) ventral anteriorly located used to attach to the columnar cells of the intestine.

Giardiasis- Summary
Giardia lamblia (also called G. intestinalis and G. duodenalis) This organism is unicellular and infection of the host results when environmentally resistant cysts are ingested. Growing, motile stages of the parasite, referred to as trophozoites, emerge from the cyst (a process called excystation) in the proximal small intestines and colonize the intestines. A certain number of these trophozoites travel to the more distal intestines and will encyst, and will be passed back into the environment in the feces of the host. The life cycle is completed when a new host ingests these cysts. This fecal-oral route of infection may result from person-to-person contact but also often involves ingesting cysts that contaminate natural waters. Because G. lamblia infects many animals in addition to human (i.e. it has many reservoir hosts), mountain and forest streams are often contaminated with cysts that are deposited there by wildlife that inhabit these areas. It is important to either filter, boil or chemically treat water from these streams before drinking it to avoid possible infection with G. lamblia.