Lab 4

assistant teacher
Tiba Noori M.
Dientamoeba fragilis
 Dientamoeba fragilis
• Despite its name, Dientamoeba fragilis is not an amoeba but an
intestinal flagellate.
• Dientamoeba fragilis is a flagellate that lacks external flagella and
therefore must be morphologically differentiated from the small
nonpathogenic amoebae (e.g., Endolimax nana, Entamoeba hartmanni
).
• Although the organism primarily moves in amoeboid fashion by
extending pseudopodia with somewhat serrated (sharp-toothed)
edges. .‫ ﻟﻴﺴﺖ أﻣﻴﺒﺎ وﻟﻜﻨﻬﺎ ﺳﻮط ﻣﻌﻮي‬Dientamoeba fragilis ‫ ﻓﺈن‬، ‫ﻋﻠﻰ اﻟﺮﻏﻢ ﻣﻦ اﺳﻤﻬﺎ‬
Dientamoeba fragilis ‫ﺷﻜﻠﻴﺎ ﻋﻦ اﻷﻣﻴﺒﺎت‬
ً ‫ﻋﺒﺎرة ﻋﻦ ﺳﻮط ﻳﻔﺘﻘﺮ إﻟﻰ اﻷﺳﻮاط اﻟﺨﺎرﺟﻴﺔ وﺑﺎﻟﺘﺎﻟﻲ ﻳﺠﺐ ﺗﻤﻴﻴﺰه‬
‫ اﻟﺼﻐﻴﺮة ﻏﻴﺮ اﻟﻤﺴﺒﺒﺔ ﻟﻸﻣﺮاض )ﻋﻠﻰ ﺳﺒﻴﻞ اﻟﻤﺜﺎل‬، Endolimax nana ، Entamoeba hartmanni
‫ﻋﻠﻰ اﻟﺮﻏﻢ ﻣﻦ أن اﻟﻜﺎﺋﻦ اﻟﺤﻲ ﻳﺘﺤﺮك ﻓﻲ اﻟﻤﻘﺎم اﻷول ﺑﻄﺮﻳﻘﺔ اﻷﻣﻴﺒﻴﺔ ﻋﻦ ﻃﺮﻳﻖ ﻣﺪ اﻷرﺟﻞ اﻟﻜﺎذﺑﺔ ﺑﺤﻮاف ﻣﺴﻨﻨﺔ‬
‫إﻟﻰ ﺣﺪ ﻣﺎ )ذات أﺳﻨﺎن ﺣﺎدة(‪.‬‬
 8-5 ‫ﻳﺤﺘﻮي ﻋﻠﻰ ﻧﻮى ﻣﻜﻮﻧﺔ ﻣﻦ‬
‫ﺷﻜﻞ ﻏﻴﺮ ﻣﻨﺘﻈﻢ‬ Shape Irregular and
Contains nuclei composed of‫ﺣﺒﻴﺒﺎت ﻣﻨﻔﺼﻠﺔ‬
amoeboid ‫و‬ amoeboid
5–8 discrete granules

.‫ﻛﺎذﺑﺔ اﻟﺰاوي إﻟﻰ واﺳﻌﺔ اﻟﻔﺼﻮص وﺷﻔﺎﻓﺔ‬

the majority of ‫ﻏﺎﻟﺒﻴﺔ اﻟﻜﺎﺋﻨﺎت‬ Pseudopodia are angular to

organisms have ‫اﻟﺤﻴﺔ‬ broad-lobed and transparent.
two nuclei, usually ‫ ﻋﺎدة ﻣﺎ‬، ‫ﻧﻮاﺗﺎن‬
joined by a spindle ‫ﻳﺘﻢ رﺑﻄﻬﻤﺎ‬
or filament ‫ﺑﻮاﺳﻄﺔ ﻣﻐﺰل أو‬
(desmose), but up ‫ﺧﻴﻮط‬ Motility Nondirectional
to 40% may be (desmos) ، ‫وﻟﻜﻦ‬
‫ﺣﺮﻛﻴﺔ ﻏﻴﺮ اﺗﺠﺎﻫﻴﺔ‬
uninucleate. ‫ﻣﺎ ﻳﺼﻞ إﻟﻰ‬40‫ ﻗﺪ‬٪
‫ﻳﻜﻮن ﻏﻴﺮ ﻧﻮوي‬.

The cytoplasm contains

ingested bacteria and other
food material.

.‫ﻳﺤﺘﻮي اﻟﺴﻴﺘﻮﺑﻼزم ﻋﻠﻰ ﺑﻜﺘﻴﺮﻳﺎ ﻣﺒﺘﻠﻌﺔ وﻣﻮاد ﻏﺬاﺋﻴﺔ أﺧﺮى‬

• In human stool specimens, D. fragilis is
almost always found as a trophozoite.
• However, the rare presence of cyst and
precyst like forms in clinical specimens
has been reported.
• Trophozoites are found in the lumen of
the large intestine.
• There‫ﺷﻜﻞ‬
is ‫ﻋﻠﻰ‬ evidence to suggest that D.
no ‫داﺋﻤﺎ‬
ً ‫ ﻓﺮاﺟﻴﻠﻴﺲ‬.‫ ﻳﻮﺟﺪ د‬، ‫ﻓﻲ ﻋﻴﻨﺎت اﻟﺒﺮاز اﻟﺒﺸﺮي‬
fragilis trophozoites invade .‫ﺗﺮوﻓﻮزوﻳﺖ‬
their
surrounding tissues.
‫ﻟﻠﻜﻴﺲ واﻷﺷﻜﺎل‬ D. fragilis
‫اﻟﻮﺟﻮد اﻟﻨﺎدر‬ ‫ﺗﻢ اﻹﺑﻼغ ﻋﻦ‬has only
‫ ﻓﻘﺪ‬، ‫ذﻟﻚ‬ ‫وﻣﻊ‬
rarely been known .‫اﻟﺴﺮﻳﺮﻳﺔ‬
to ‫اﻟﻌﻴﻨﺎت‬
ingest ‫اﻟﻜﻴﺴﺎت ﻓﻲ‬
red blood‫اﻟﺘﻲ ﺗﺸﺒﻪ‬
.‫ ﻓﻲ ﺗﺠﻮﻳﻒ اﻷﻣﻌﺎء اﻟﻐﻠﻴﻈﺔ‬Trophozoites ‫ﺗﻢ اﻟﻌﺜﻮر ﻋﻠﻰ‬
cells.
‫ﻧﺎدرا ﻣﺎ ﻋُ ﺮف ﻋﻦ ﺑﻜﺘﻴﺮﻳﺎ ‪ D. fragilis‬أﻧﻬﺎ‬
‫ﻻ ﻳﻮﺟﺪ دﻟﻴﻞ ﻳﺸﻴﺮ إﻟﻰ أن ‪ D. fragilis trophozoites‬اﻟﻤﺤﻴﻄﺔ ﺑﺎﻷﻧﺴﺠﺔ‪ً .‬‬
‫ﺗﺘﻨﺎول ﺧﻼﻳﺎ اﻟﺪم اﻟﺤﻤﺮاء‪ .‬ﻏﺰو ﺑﻬﻢ‬
 Transmission
• fecal-oral transmission routes.
• Transmission via helminth eggs (e.g., via Enterobius sp.
eggs). Enterobius sp. (pinworm) eggs
Ascaris egg
.‫*ﻃﺮق اﻧﺘﻘﺎل اﻟﺒﺮاز اﻟﻔﻤﻮي‬
‫•اﻻﻧﺘﻘﺎل ﻋﺒﺮ ﺑﻴﺾ اﻟﺪﻳﺪان اﻟﻄﻔﻴﻠﻴﺔ‬
Enterobius ‫ ﻋﺒﺮ‬، ‫)ﻋﻠﻰ ﺳﺒﻴﻞ اﻟﻤﺜﺎل‬
(‫ ﺑﻴﺾ‬.sp
 Clinical Presentation ‫اﻟﻌﺮض اﻟﺴﺮﻳﺮي‬

• Infection appears to be more common in children.

• Both asymptomatic and symptomatic infection
• particular strains of D. fragilis are associated with symptomatic
infection in humans.
• The reported clinical manifestations have sometimes been
described as similar to those of colitis , appendicitis
.‫اﻷﻃﻔﺎل‬ ‫أﻛﺜﺮ ﺷﻴﻮﻋً ﺎ ﻋﻨﺪ‬, ‫اﻟﻌﺪوى‬
or irritable
‫ﻳﺒﺪو أن‬
.D ‫ﻣﻌﻴﻨﺔ ﻣﻦ‬
bowel ‫ﺗﺮﺗﺒﻂ ﺳﻼﻻت‬
syndrome . ‫ﻛﻞ ﻣﻦ اﻟﻌﺪوى اﻟﻤﺼﺤﻮﺑﺔ ﺑﺄﻋﺮاض أو اﻟﻌﺪوى اﻟﻤﺼﺤﻮﺑﺔ ﺑﺄﻋﺮاض‬
• gastrointestinal symptoms of pain, diarrhea, as well as nausea and
fatigue.
• intense diarrhea accompanied with blood or mucus, abdominal
pain.
‫*ﺗﻢ وﺻﻒ اﻟﻤﻈﺎﻫﺮ اﻟﺴﺮﻳﺮﻳﺔ اﻟﻤﺒﻠﻎ ﻋﻨﻬﺎ أﺣﻴﺎ ًﻧﺎ ﺑﺄﻧﻬﺎ ﻣﺸﺎﺑﻬﺔ ﻟﺘﻠﻚ اﻟﺨﺎﺻﺔ ﺑﺎﻟﺘﻬﺎب اﻟﻘﻮﻟﻮن أو اﻟﺰاﺋﺪة‬
‫اﻟﺪودﻳﺔ أو ﻣﺘﻼزﻣﺔ اﻟﻘﻮﻟﻮن اﻟﻌﺼﺒﻲ‪.‬‬
‫* أﻋﺮاض اﻟﺠﻬﺎز اﻟﻬﻀﻤﻲ ﻣﻦ اﻷﻟﻢ واﻹﺳﻬﺎل‬
‫وﻛﺬﻟﻚ اﻟﻐﺜﻴﺎن واﻟﺘﻌﺐ‪.‬‬
‫*إﺳﻬﺎل ﺷﺪﻳﺪ ﻣﺼﺤﻮب ﺑﺪم أو ﻣﺨﺎط وآﻻم ﻓﻲ اﻟﺒﻄﻦ‪.‬‬
 ‫اﻟﻌﻴﻨﺎت‬
Samples
• Stool sample for microscopic and antigen detection method.
• Serum for serology.
• Multiple samples may be necessary to rule out the presence of this
organism because the amount of parasite shedding may vary from
day to day.
.‫*ﻋﻴﻨﺔ اﻟﺒﺮاز ﻟﻄﺮﻳﻘﺔ اﻟﻜﺸﻒ اﻟﻤﺠﻬﺮي وﻣﺴﺘﻀﺪ‬
.‫*ﻣﺼﻞ اﻷﻣﺼﺎل‬
‫*ﻗﺪ ﺗﻜﻮن اﻟﻌﻴﻨﺎت اﻟﻤﺘﻌﺪدة ﺿﺮورﻳﺔ ﻻﺳﺘﺒﻌﺎد وﺟﻮد ﻫﺬا اﻟﻜﺎﺋﻦ اﻟﺤﻲ ﻷن ﻛﻤﻴﺔ اﻟﻄﻔﻴﻞ ﻗﺪ‬
.‫ﺗﺨﺘﻠﻒ ﻣﻦ ﻳﻮم ﻵﺧﺮ‬
 ‫اﻟﺘﺸﺨﻴﺺ‬
Laboratory Diagnosis ‫اﻟﻤﺨﺘﺒﺮي‬

• Microscopically:
• the trophozoite is the only stage found in stools of infected individuals.
• Wet mount: Unless the specimen is extremely fresh, simple wet film
examination of a faecal sample is unlikely to reveal many cases of
infection,
• Permanent staining: stained with trichrome or iron haematoxylin or
another permanent stain. :‫ﻣﺠﻬﺮﻳً ﺎ‬
.‫اﻟﻤﺼﺎﺑﻴﻦ‬
• The stain of choice ‫اﻟﻤﻮﺟﻮدة ﻓﻲ ﺑﺮاز اﻷﻓﺮاد‬
for distinguishing ‫اﻟﻤﺮﺣﻠﺔ اﻟﻮﺣﻴﺪة‬
the individual ‫ ﻫﻲ‬trophozoite
chromatin granules‫إن‬is
iron hematoxylin.
‫ ﻓﻤﻦ ﻏﻴﺮ اﻟﻤﺮﺟﺢ أن ﻳﻜﺸﻒ ﻓﺤﺺ اﻟﻔﻴﻠﻢ اﻟﺮﻃﺐ‬، ‫ ﻣﺎ ﻟﻢ ﺗﻜﻦ اﻟﻌﻴﻨﺔ ﻃﺎزﺟﺔ ﻟﻠﻐﺎﻳﺔ‬:‫اﻟﺘﺮﻛﻴﺐ اﻟﺮﻃﺐ‬
• Dientamoeba fragilis trophozoites can‫ﺣﺎﻻت‬
، ‫اﻟﻌﺪوى‬ easily be ‫ﻋﻦ‬overlooked
‫اﻟﻌﺪﻳﺪ ﻣﻦ‬ or
‫اﻟﺒﺴﻴﻂ ﻟﻌﻴﻨﺔ اﻟﺒﺮاز‬
misidentified because they are pale-staining and their nuclei sometimes
resemble those of Endolimax nana or Entamoeba hartmanni.
 ‫ﺗﻠﻄﻴﺦ داﺋﻢ‪ :‬ﻣﻠﻄﺦ ﺑﺜﻼﺛﻲ اﻟﻜﺮوم أو ﻫﻴﻤﺎﺗﻮﻛﺴﻴﻠﻴﻦ اﻟﺤﺪﻳﺪ أو ﺻﺒﻐﺔ داﺋﻤﺔ أﺧﺮى‪.‬‬
‫وﺻﻤﺔ ﻋﺎر اﻻﺧﺘﻴﺎر ﻟﻠﺘﻤﻴﻴﺰ ﺑﻴﻦ ﺣﺒﻴﺒﺎت اﻟﻜﺮوﻣﺎﺗﻴﻦ اﻟﻔﺮدﻳﺔ ﻫﻲ ﻫﻴﻤﺎﺗﻮﻛﺴﻴﻠﻴﻦ اﻟﺤﺪﻳﺪ‪.‬‬

‫ﻳﻤﻜﻦ ﺑﺴﻬﻮﻟﺔ اﻟﺘﻐﺎﺿﻲ ﻋﻦ ﻋﻨﺎﺻﺮ ‪ Dientamoeba fragilis trophozoites‬أو اﻟﺘﻌﺮف ﻋﻠﻴﻬﺎ ﺑﺸﻜﻞ‬
‫ﺧﺎﻃﺊ ﻷﻧﻬﺎ ذات ﺑﻘﻊ ﺑﺎﻫﺘﺔ وﺗﺸﺒﻪ ﻧﻮاﺗﻬﺎ أﺣﻴﺎ ًﻧﺎ ﺗﻠﻚ اﻟﻤﻮﺟﻮدة ﻓﻲ ‪ Endolimax nana‬أو‬
‫‪.Entamoeba hartmanni‬‬
 Binucleate form of a trophozoite of D. fragilis, stained with
trichrome.
The two small nuclei (arrows) have fragmented karyosomes
containing discrete chromatin granules (darts).
‫ ﺗﺤﺘﻮي‬.‫ ﻣﻠﻄﺦ ﺑﺜﻼﺛﻴﺔ اﻟﻜﺮوم‬، D. fragilis ‫ ﻟـ‬trophozoite ‫ﺷﻜﻞ ﺛﻨﺎﺋﻲ اﻟﻨﻮاة ﻣﻦ‬
‫اﻟﻨﻮاﺗﺎن اﻟﺼﻐﻴﺮﺗﺎن )اﻷﺳﻬﻢ( ﻋﻠﻰ ﻧﻮاة ﻣﺠﺰأة ﺗﺤﺘﻮي ﻋﻠﻰ ﺣﺒﻴﺒﺎت ﻛﺮوﻣﺎﺗﻴﻦ‬
(‫ﻣﻨﻔﺼﻠﺔ )ﺳﻬﺎم‬
Uninucleate form of a trophozoite of D. fragilis,
stained with trichrome.
.‫ ﻣﻠﻄﺦ ﺑﺎﻟﺘﺮﻳﻜﺮوم‬، D. fragilis ‫ ﻣﻦ‬trophozoite ‫ﺷﻜﻞ ﻏﻴﺮ ﻧﻮوي ﻣﻦ‬
Binucleate and
uninucleate
forms of
trophozoites of D.
fragilis, stained
with trichrome.
‫اﻷﺷﻜﺎل ﺛﻨﺎﺋﻴﺔ اﻟﻨﻮاة وﻏﻴﺮ اﻟﻨﻮى ﻣﻦ‬
، D. fragilis ‫ ﻣﻦ‬trophozoites
.‫ﻣﻠﻄﺨﺔ ﺑﺎﻟﺘﺮﻳﻜﺮوم‬
 :‫ﻃﺮﻳﻘﺔ اﻟﺘﺮﻛﻴﺰ‬
Concentration method:
• The trophozoite stage of the parasite is not usually detectable if stool
concentration methods are used.
• concentration methods will certainly destroy the trophozoites.

Molecular methods:
.‫ﻃﺮق ﺗﺮﻛﻴﺰ اﻟﺒﺮاز‬ ‫ﻻ ﻳﻤﻜﻦ ﻋﺎدة اﻛﺘﺸﺎف ﻣﺮﺣﻠﺔ اﻟﻄﻔﻴﻠﻲ ﻣﻦ اﻟﻄﻔﻴﻠﻲ إذا ﺗﻢ اﺳﺘﺨﺪام‬

More recently, both conventional and real-time .‫ﻃﺮق اﻟﺘﺮﻛﻴﺰ ﺳﺘﺪﻣﺮ ﺑﺎﻟﺘﺄﻛﻴﺪ اﻟﺠﻮاﺋﺰ‬
polymerase chain reaction (RT-PCR) methods have
been used to diagnose D. fragilis in patients. A
recent study evaluated methods of detection for D.
fragilis and RT-PCR was shown to be the most
sensitive of all diagnostic methods.
 ‫اﻟﻄﺮق اﻟﺠﺰﻳﺌﻴﺔ‪:‬‬
‫ﻓﻲ اﻵوﻧﺔ اﻷﺧﻴﺮة ‪ ،‬ﺗﻢ اﺳﺘﺨﺪام ﻛﻞ ﻣﻦ ﻃﺮق ﺗﻔﺎﻋﻞ اﻟﺒﻠﻤﺮة اﻟﻤﺘﺴﻠﺴﻞ‬
‫اﻟﺘﻘﻠﻴﺪﻳﺔ وﻓﻲ اﻟﻮﻗﺖ اﻟﺤﻘﻴﻘﻲ )‪ (RT-PCR‬ﻟﺘﺸﺨﻴﺺ داء اﻟﺒﻠﻤﺮة اﻟﻬﺸﺔ ﻓﻲ‬
‫اﻟﻤﺮﺿﻰ‪ .‬أﻇﻬﺮت دراﺳﺔ ﺣﺪﻳﺜﺔ ﻗﻴﻤﺖ ﻃﺮق اﻟﻜﺸﻒ ﻋﻦ ‪ D. fragilis‬و‬
‫‪ RT-PCR‬أﻧﻬﺎ اﻷﻛﺜﺮ ﺣﺴﺎﺳﻴﺔ ﻣﻦ ﺑﻴﻦ ﺟﻤﻴﻊ ﻃﺮق اﻟﺘﺸﺨﻴﺺ‪.‬‬
 Antigen detection: ‫ﻛﺸﻒ اﻟﻤﺴﺘﻀﺪ‬

‫• ﻃﺮﻳﻘﺔ واﻋﺪة ﻟﻠﺘﺄﻟﻖ اﻟﻤﻨﺎﻋﻲ ﻏﻴﺮ اﻟﻤﺒﺎﺷﺮ ﻟﻠﻜﺸﻒ ﻋﻦ اﻟﻨﻮاﺷﻂ ﻓﻲ ﻋﻴﻨﺎت اﻟﺒﺮاز اﻟﺜﺎﺑﺘﺔ‬

• A promising indirect immunofluorescence method for the detection

of trophozoites in fixed faecal specimens

‫ﻛﺸﻒ اﻻﺟﺴﺎم اﻟﻤﻀﺎده‬ Antibody detection:

• A very high proportion of healthy children are apparently seropositve
making serology of little use for identifying infected patients.

‫• ﻳﺒﺪو أن ﻧﺴﺒﺔ ﻋﺎﻟﻴﺔ ﻣﻦ اﻷﻃﻔﺎل اﻷﺻﺤﺎء ﻳﻌﺎﻧﻮن ﻣﻦ اﻹﻣﺴﺎك اﻟﻤﺼﻠﻲ ﻣﻤﺎ ﻳﺠﻌﻞ‬
.‫اﺳﺘﺨﺪام اﻷﻣﺼﺎل ﺿﺌﻴﻼً ﻓﻲ ﺗﺤﺪﻳﺪ اﻟﻤﺮﺿﻰ اﻟﻤﺼﺎﺑﻴﻦ‬