UERMMMCI: CAHP (BATCH 2019)  Two Developmental stages of ciliates and flagellates in GIT

PARASITOLOGY – FINALS - Cyst

Reference: Lecture notes prepared by Prof. Eduardo - Infective stage to man
EBON 2017 - Sturdy – able to survive in chlorinated water
- Trophozoites
CILIATES AND FLAGELLATES - Pathogenic stage
Outline: - Can‟t survive outside environment for long
period of time
I. Flow Cyst  trophozoite EXCYSTATION
II. Introduction Trophozoites  cyst ENCYSTATION
III. Balantidum coli
IV. Mastigophora flagellates Balantidum coli
- Giardia Intestinalis / Giardia duodenalis / INTRODUCTION
lambia  Parasites of pig but can cause disease to man
- Chilomastix mesnili  1857 – Malmstem
V. Trichomonads - First to recognized B. coli in 2 humans with dysentery
- Dientamoeba fragilis (blood and mucus in stool)
- Trichomonas vaginalis - First name given: “Paramecium coli” due to several cilia
- Trichomonas tenax  1861 – Leuckart
- Trichomonas hominis - First saw the morphology similarly with that parasites
from pig‟s intestine
Flow  STEIN
- Renamed: “balantidium coli”I looks like a bag with furs
Introduction - “balanto” – greek word “bag”
 Habitat: Colon, Cecum, LI
Morphology  Disease: Balantidiasis
 Opportunistic pathogen
Life Cycle - Pathophysiology

MORPHOLOGY
Laboratory Diagnosis
1. Trophozoite
 Tissue invader
Epidemiology
 30-150 micrometer in length
PVC  Cy (cystosome or mouth)
 Micronucleus - reproduction
INTRODUCTION  Macronucleus
Recall: - metabolism and growth
- „sausage shape‟ tapered anterior end surface
 Sarcodina – Amoeba covered with cilia
 Mastigophora – Flagellates  CV (contractile vacuole)
 Ciliophora – Ciliates
 Apicomplexa – Sporozoa, Ciccidians
 Sporozoa
- e.g. Malarial parasite
 Coccidians
- e.g. Toxoplasma & Cryptosporidia

 Types of Nucleus of Protozoa

- Entamoeba histolytica
- Pseudopod
- Karyosome (nucleolus)
- Nuclear chromatin (chromatin granules) Figure 1. B. coli trophozoite (Image obtained from
- “fine chromatin granules with small central www.cdc.gov/dpdx)
granules” 2. Cysts
- Entamoeba coli  Size: 45-65 micrometer
- Larger chromatin granules than E. histolytica  Micronucleus - reproduction
- Eccentric karyosome  Macronucleus - metabolism and growth
- G. lamblia  CV (contractile vacuole)
- “large central karyosome”  Without cilia
- D. fragilis
- ”clusters of 4-8 chromatin granules”

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 LABORATORY DIAGNOSIS up to EPIDEMIOLOGY
 Specimen: stool
 Test: concentration wet preparation to visualize the cilia.(s.a
Zinc flotation method
 Epdiemiology
- Areas with poor sanitation
- Close contact with pigs or pig‟s feces
- Overcrowded institution (e.g. asylum, prisons,
orphanage)
- Bolivia
- Papua New Guinea
- Philippines In 2005, a study was done on pigs in southern
Figure 2. Cysts of B. coli (Image obtained from
& northern PH. Revealed 66.1% prevalence of B. coli
www.cdc.gov/dpdx)
infection in pigs‟
LIFE CYCLE
Cecum & colon Mastigophora (Flagellates)
 Characteristics:
- Flagella (one or more) move in “counter clockwise”
Invade mucosa & multiply by binary fission or conjugation
fashion creating a rapid movement.
- All reproduce ASEXUALLY (binary fission)
Secretion of hyaluronidase then digests epithelium causing
Ulceration of the intestinal mucosa/
 Members:
- Chilomastix mesnii
Encystation in rectum
- Giardia duodenalis
- Trichomonads:
Ingestion of contaminated water excystation in small intestine\
o Dientamoeba fragilis
o Trichomonas spp
PATHOPHYSIOLOGY
 Characteristics of ulcer produced: - Trypanosoma spp
- Larger compared to amoeba - Leishmania spp
- Multiple
- Wide neck  Classification:
- Round or oval base - Blood and tissue
 Incubation period 4 to 5 days for Balantidiasis o Leishmania & trypanosoma
 S/Sx: - Intestinal
- Watery stool or with blood and pus o G. dudodenalis & C. mesnili
- Nausea - Trichomonads
- Anorexia o D. fragilis, T. vaginalis
- Tenesmus
- Abdominal pain Giardia intestinalis / Giardia duodenalis or Giardia lambia
INTRODUCTION up to MORPHOLOGY
- Diarrhea
 History:
 Clinical manifestations:
- 1859 – first describe by Lamb
o 80% of infected are asymptomatic carriers (serve as
 The father of microbiology – Anton Van leeuwenhoek
reservoir)
- 1681 – discovered G. duodenalis in his stool
o Chronic-colitis – non bloody diarrhea, abdominal
 Disease: Giardiasis
cramps, halitosis
o Fulminating balantidiasis – mucoid and bloody stool,
1. Trophozoites
weight loss
 12-15 micrometer length
 Tissues invaded
 2 nuclei with large karyosome
- Mesenteric nodes
 Medial bodies
- Appendix
 Axoneme
- Cecum (habitat)
 2 ventral flagella
- Extraintestinal
 Adhesive disc/sucking disc
- Lungs
 4 pairs of flagella
- Liver
 2 anterior
- GU
 2 posterior
 Possible pathway to the lungs
 2 caudal
- Lymphatic system
 2 ventral
- Hepatic portal circuit
o As the trophozoites mature to cyst the flagella
- Perforation of the colon and spread thru the peritoneal
and sucking discs are lost.
cavity
 “Smiley face”
Note: the presence of cilia in the lungs, will challenge you, thus,
 Motility: “falling leaf” motion
examine accurately

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 LABORATORY DIAGNOSIS
 Specimen: stool
 3 stool sample taken every 2 days interval
 Look for Charcot-leyden crystals (products from eosinophils)

Figure 3. Trophozoites of G.lambia ((Image obtained from

www.cdc.gov/dpdx)

LIFE CYCLE
Stomach  gastric acid stimulates excystation
 IFA, ELISA
Attached to intestinal villi (duodenum and jejunum) via sucking  String test (Entero test)
disc - Use a gelatin capsule with an attached thread
- One end is attached to the inner aspect of the
Will cause  Flattening of villi, crypt hyperplasia, decrease patient‟s cheek
lactase enzyme, bile salts taken up for excystation and bacterial - The capsule is swallowed
growth - Thread is withdrawn and shaken in saline to release the
trophozoite.
Encystation in large intestines
EPIDEMIOLOGY
PATHOPHYSIOLOGY  Reservoir host:
 Flattening of villi lead to decreased in absorption - Human
 Crypt hyperplasia associated with increased enterocyte - Dogs
migration rate - Beaver
- Can lead to lactose deficiency.  Vehicles:
- Extremely large no. of trop may lead to direct physical - Water
blockage of nutrient uptake esp fat soluble substances - Resistant traditional 5%chlorine
(Vitamin B12)  Malabsorption syndrome - Survive at 62C, survive in lakes (3 months)
 Clinical manifestations - Uncooked contaminated vegetables and fruits
- 50% of infected are asymptomatic carriers.  Worldwide
- Acute giardiasis: less nutrient and H20 absorption >  Philippines – 1.6 to 22%
“traveler” diarrhea, flatulence with odor of rotten egg-  The first published study showed that 86% of
like smell (1-2 weeks) isolates belongs to genotype assemblage B.
- Chronic giardiasis – depends on number of  Results of experiment done in Animals
trophozoites (diarrhea, malabsorption syndrome, fatty - Malabsorption  caused diarrhea not active secretion
stool) e.g. enterotoxin of bacteria
Crypt and villus astrophy  lactase deficient  Vitro
 - Human milk  kills trophozoites
Available lactose used for bacterial growth
 Chilomastix mesnili
Bacterial growth & excystation used bile salt MORPHOLOGY
 1. Cysts
Decreased in bile salt  6-10 micrometer
  Thick cyst wall
Less uptake of lipid  Cystosome (“open safety pin” or “shepherds crook”
  Lemon shape with hyaline protrusion
Steatorrhea  Large nucleus with central karyosome
 Host immunity
- Mostly humoral
- Due to the exhibition of Ag variation
- Expressed 180 different Antigens over 6-12 generations
IgM & sIgA plays important role Patient with sIgA
deficiency + with duodenal or jejunal diverticula
(congenital) = chronic giardiasis

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  Life cycle:
- Proposed transmission: via helminths ova Enterobius
and Ascaris
- Ingestion
- Binary fission
- Trophozoite in the feces
- Cycle continue
 Clinical manifestations:
- Diarrhea
- Abdominal pain
- Anorexia
- Fatigue
Figure 4. Cyst of Chilomastix mesnili (Image obtained from
- Probably acts as low-grade irritant of intestinal mucosal
www.cdc.gov/dpdx)
surfaces that may lead to inflammation.
2. Trophozoites:
- 1977 – 1st case reported in Thailand
 12-20 micrometer
 Laboratory Diagnosis
 Pear shape
 Difficult to identify in fresh wet mount can be mistaken
 3 flagella
with
 1 flagellum (longer)
 WBC
 Large nucleus with central karyosome located
 Faecal debris
anteriorly
 Intestinal cells
 Cytostome (“ open safety pin” & “sheperds crook”)
o Formed stool – trophozoite not motile
 Spiral groove
o Watery stool – actively motile

Figure. 5 Trophozoite of Chilomastix mesnili (Image obtained from

www.cdc.gov/dpdx)

Trichomonads
 Characteristics: Figure 6. Trophozoite of Dientamoeba fragilis (Image obtained
- Anaerobic from www.cdc.gov/dpdx)
- Cyst not formed
- Transmission: direct contact
- Possess 4-6 flagella at the apical pole
- Axostyle present

Dientamoeba fragilis
INTRODUCTION up to LABORATORY DIAGNOSIS
 Di – 2
 Ent – intestine
 Amoeba – with pseudopod
 Fragilis – fragile
 Formerly under Family Endamoebidae
 Habitat: cecum & upper colon
 Structure of Trichomonas
 Reasons for transfer to Family Trichomonads:
- Four anterior flagella (jerky motility)
 Structures:
- Nucleus
- Has different nucleus
- Axostyle (central rod)
- Exist as trophozoite only
- Posterior flagellum (T. hominis)
- Has extra spindle
- Undulating membrane
- Closely related to Genus Histomonas (
Amoeboflagellates)
Trichomonas tenax (tetra trichomonas buccalis)
 Characteristics
 Commensals in mouth of humans and pirates
- No cystic stage
 Found in tartar, defect of caries? Teeth
- 5-15 micrometer
- 2 nuclei
- Clusters of 4-8 chromatin granules
- Uses trichrome strain

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 Trichomonas vaginalis - Vulva itching or swelling
MORPHOLOGY up to LIFE CYCLE - Burning sensation/vaginal itching (15-20%)
 Nucleus - Abundant “frothy” yellow green vaginal discharge (50-
 Flagella 75%)
 Undulating membrane (half of the body) - Punctate cervical microhemorrhages (25%)
 Life cycle:
Female:  Complications
- Vagina - Premature delivery due to premature rupture of the
- Skene‟s gland (urethra) membrane (PPROM)
Male: - Baby may have low birth weight (<5.5 lbs. at birth )
- Urethra - Predisposing factors to:
 Note: Pass thru sexual intercourse with multiple partners.  HIV infection (presence of open
- Prefers alkaline pH wound)
- Can live outside the host up to 30 minutes  Cervical cancer (2001)
- May survive in a host (esp. men) >two years
- Menopause (6-7 pH) LABORATORY DIAGNOSIS up to PVC
- Use of birth control pills  Pelvic examination
- Prolonged used of antibiotic. - “strawberry cervix” vaginal wall have erythema and
punctuate hemorrhages
- T. vaginalis adhere to vaginal squamous epithelial.
“Strawberry cervix” is due to extensive inflammation
and erosion of vaginal or urethral epithelium
- N. gc adhere to endocervix columnar epithelium
Methods of specimen collection
Female:
- Self obtained low vaginal swab (SOLV)
- High vaginal swab
Male:
- Get the scanty transparent or white
Figure 6. Trophozoite of Trichomonas vaginalis (Image obtained discharge
from www.cdc.gov/dpdx)  Treatment:
o Sometimes men may need no treatment as the
infection is likely to resolve itself in a couple of weeks.
CHARACTERISTICS
Characteristics: o Metronidazole one megadose taken orally
 “Clean-up organism” o Tinidazole
 Feeds on: o Most curable STI
- Cell debris o Pregnant:
- Pus - 1st trimester – tropical clotrimazole
- Bacteria - 2nd and 3rd trimester – metronidazole orally
o Home remedies:
PATHOGENESIS - Sitz bath with lemon and vinegar
 Incubation period: 4 to 28 days - Lactobacillus acido good bacteria
 PVC:
Adhere to vaginal squamous epithelial cell and release protein - Practice monogamy
enzyme (digest tissue) - Avoid intercourse during treatment
 - Notify his or her partner so they can be treated to avoid
Triggers: extensive inflammation and erosion of vaginal or re-infection.
urethral epithelium - Avoid unprotected sexual contact (condoms reduce
 risk only, not 100% protective)
Effects: redness, swelling, pain, itching, pus, exudation. - Take shower after intercourse
- Don‟t share towels or underwears.
 Symptoms: in male - Frequently co-exist with other STI
- Mild white or transparent discharge from the penis
(50-60%)
- Itching of the urethra (25%)
- Burning sensation after urination or ejaculation (12-
25%)
- Holds infections for years, asymptomatic (50-90%)
- Some may develop:
 Prostitis
 Epididymitis
 Symptoms: in female
- Discomfort or pain during intercourse

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