Entamoeba histolytica

2 years ago by Dr.E.I 0

General Description
Entamoeba histolytica is an ameba that feeds on cells in the human colon. It is the
cause of amebic dysentery (bloody diarrhea) as well as colonic ulcerations. The
infection is also referred to as amebiasis. If the organisms spread throughout the
body via the bloodstream they may cause abscesses in the liver or, less frequently,
other organs.
Morphology
The organism has two forms.
The trophozoite is 10 to 60 µm in diameter, ameboid, actively motile, and often
erythrophagocytic. In stained specimens, the nucleus has a central karyosome with
finely beaded peripheral chromatin.
E.histolytica trophozoite

E.histolytica trophozoite
The Cyst form
Encystation begins with the trophozoites become more spherical and the
appearance of chromatoid bodies in the cytoplasm. Chromatoid bodies (cb) are
stained elongated structures with round ends and represent the aggregation of
ribosomes. The cyst wall is composed of chitin and has a smooth refractile
appearance. Cyst maturation involves two rounds of nuclear replicationwithout cell
division and cysts with 1-4 nuclei (Nu) are found in feces. Sometimes the young
cysts (ie, 1-2 nuclei) will have a glycogen vacuole (vac) which will appear as a
clear area in stained specimens. This vacuole will sometimes displace and alter the
morphology of the nuclei. The chromatoid bodies tend to disappear as the cyst
matures. The cysts are generally 12-15 µm in diameter. Cysts are immediately
infective upon excretion with the feces and will be viable for weeks-to-months
depending on environmental conditions.

E.histolytica cyst
 e.histolytica cyst and trophozite

Transmission
E. histolytica is spread by the fecal-oral route. This is achieved through food or
water contaminated with cysts, oral-anal sexual contact, or occasionally directly in
childcare centers or institutions for the developmentally challenged. The disease is
found far more frequently in people from developing countries or travelers to such
areas than in developed countries.
Pathogenesis & Clinical Manifestations
Pathogenic and non-pathogenic strains of E histolytica inhabit the human
digestive tract. Even pathogenic strains may live in the lumen as benign
commensals.
If mucosal invasion occurs, it may be limited to a few simple superficial
erosions or it may progress to total involvement of the colonic mucosa with
ulceration. The clinical manifestations vary with the extent of involvement.
Mucosal erosion causes diarrhea, which increases in severity with increasing
area and depth of involvement.
Symptoms are also affected by the site of the infection. The more distal
the lesion in the colon, the greater the likelihood and severity of
symptoms; thus small rectal lesions are more likely to be symptomatic
than larger cecal lesions.
Rectal bleeding is only slightly less common than diarrhea and is
usually, but not invariably, associated with diarrhea. Such bleeding
may be grossly apparent or may be occult and demonstrable only by
chemical testing for blood. Urgency, tenesmus, cramping abdominal
pain and tenderness may be present.
Amebic dysentery has a dramatically different clinical presentation. The
diarrhea is replaced by dysenteric stools consisting largely of pus and
blood without feces. There is evidence of systemic toxicity with fever,
dehydration, and electrolyte abnormalities. Tenesmus and abdominal
tenderness are regular features. This fulminant presentation may occur
suddenly or evolve from less severe, pre-existing disease.
Occasionally, and for no apparent reason, colonic infection with E
histolytica will evoke a proliferative granulomatous response at an ulcer site.
This infectious pseudotumor, called an ameboma, may become the leading
point of an intussusception or may cause intestinal obstruction. This
complication is uncommon.
Extraintestinal amebiasis begins with hepatic involvement. Many patients with
acute intestinal infection also have hepatomegaly, but in these cases amebas
are not demonstrable in the liver and the pathogenesis of this hepatomegaly is
not clear.
A focal amebic abscess in the liver represents metastasis from
intestinal infection. Symptomatic intestinal infection need not be
present. The abscess appears as a slowly enlarging liver mass. Often
the patient will have right upper quadrant pain, which may be referred
to the right shoulder. If the abscess is located in a palpable portion of
the liver, the area will be tender.
Occasionally the enlarging abscess presses on the common bile duct
and causes jaundice. If located under the dome of the diaphragm, the
abscess may cause elevation of the dome of the diaphragm which
presses on the right lung base, causing atelectasis and physical
findings of consolidation. As the abscess nears the diaphragm the
inflammation may stimulate pleural effusion.
Pleural, pulmonary, and pericardial infection occurs as a result of direct
extension from the liver. Lung involvement is far more common than
pericardial infection. Infection metastatic from the liver can involve
other viscera or can give rise to a brain abscess. However, these
complications are uncommon
 E.histolytica phathogenisis

Diagnosis

Microscopic techniques employed in a diagnostic clinical laboratory include wet

preparation, concentration, and permanently stained smears for the identification of E.
histolytica in feces.
Microscopic examination of a direct saline (wet) mount is a very insensitive method which
is performed on a fresh specimen.
The sample should be examined within 1 h of collection to search for motile
trophozoites which may contain RBCs. However, in patients who do not present
with acute dysentery, trophozoites will not contain RBCs.
Patients with asymptomatic carriage generally have only cysts in the fecal sample.
Although the concentration technique is helpful in demonstrating cysts, the use of
permanently stained smears (trichrome or iron hematoxylin) is an important method for
recovery and identification of Entamoeba species.
As Entamoeba trophozoites generally degenerate rapidly in unfixed fecal specimens and
refrigeration is not recommended, specimens should be preserved with a fixative which
prevents the degradation of the morphology of the parasite and allows concentration and
permanent smears to be performed.
Fixatives used for the concentration procedure include Schaudinn’s fluid,
merthiolate iodine-formalin, sodium acetate-acetic acid-formalin (SAF), or 5% or
10% formalin. The fixatives for the permanently stained smears include trichrome,
iron hematoxylin, Ziehl-Neelsen stains, modified polyvinyl alcohol (PVA)
(containing mercury compounds), and SAF.
Culture Methods
Culture techniques for the isolation of Entamoeba species have been available for
over 80 years. Culture media include xenic (diphasic and monophasic) and axenic
systems. Xenic cultivation is defined as the growth of the parasite in the presence
of an undefined flora Different monophasic media that were developed for E.
histolytica are the egg yolk infusion medium of Balamus
Antibody detection methods
Many different assays have been developed for the detection of antibodies, including
indirect hemagglutination (IHA), latex agglutination, immunoelectrophoresis,
counterimmunoelectrophoresis (CIE), the amebic gel diffusion test, immunodiffusion,
complement fixation, indirect immunofluorescence assay (IFA), and enzyme-linked
immunosorbent assay (ELISA). A variety of antibody assays for detection of E.
histolytica antibodies in human serum are also commercially available

E.histolytica cysts in wet mount

Entamoeba histolytica/dispar. Formalin-fixed specimen. An immature cyst with two nuclei but without a vacuole.

e.histolytica cyst in iodine mount

 Entamoeba histolytica. Amoebic dysentery. Wet mount examination (unfixed specimen). Numerous hematophagous
trophozoites. The nucleus is never seen in this kind of preparation.

Entamoeba histolytica/dispar. SAF-fixed specimen, wet mount examination. Above, an elongated trophozoite with two nuclei
(telophase). Below, a rounded one (pre-cystic form) with dot-like karyosome

treatment
Intestinal Infection: Usually nitromidazole derivatives are used because they are highly effective against the trophozoite
form of the amoeba. Since they have little effect on amoeba cysts, usually this treatment is followed by an agent (such as
paromomycin or diloxanide furoate) that acts on the organism in the lumen.

Liver abscess: In addition to targeting organisms in solid tissue, primarily with drugs like mitronidazole and chlorquine,
treatment of liver abscess must include agents that act in the lumen of the intestine (as in the preceding paragraph) to
avoid re-invasion. Surgical drainage is usually not necessary except when rupture is imminent.
Comparación e. coli y e. harmanni

ENTAMOEBA HARTMANNI
Entamoeba coli-trofozoitos
Entamoeba coli is a non-pathogenic species of Entamoeba that frequently exists as a commensal
parasite in the human gastrointestinal tract. 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.[1] This
amoeba does not move much by the use of its pseudopod, and creates a "sur place (non-
progressive) movement" inside the large intestine. Usually, the amoeba is immobile, and keeps its
round shape. This amoeba, in its trophozoite stage, is only visible in fresh, unfixed stool
specimens. Sometimes the Entamoeba coli have parasites as well. One is the fungus Sphaerita spp.
This fungus lives in the cytoplasm of the E. coli.[2] 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.[3] The scientific name of the amoeba, E. coli, is often
mistaken for the bacterium, Escherichia coli. Unlike the bacterium, the amoeba is mostly harmless,
and does not cause as many intestinal problems as some strains of the E. coli bacterium. Some of
these harmful strains are inside raw or uncooked meat that is consumed. For example, the
bacterium, E. coli O157:H7, which can cause illness, and even death, if eaten. To make the naming
of these organisms less confusing, "alternate contractions" are used to name the species for the
purpose making the naming easier; for example, using Esch. coli and Ent. coli for the bacterium
and amoeba, instead of using E. coli for both. Entamoeba species all come in monogenetic forms,
or having one generation lifecycles. E. coli has "three distinct morphological forms exist airing the
life cycle-Trophozoite, Pre-cystic stage and Cystic stage." This lifecycle gives raise to the general
way of how Entamoeba species forms. This parasite has one large nucleus with a thick membrane
surrounding the nucleus. There are many chromatin inside the nucleus, and one large, irregular-
shaped karyosome.[8] The chromatin is clumped, and uneven in disperse inside the nucleus. The
parasite forms by binary fission like most Entamoeba spp.[9] The mature cyst is the infective stage,
and is known to survive longer than those of E. histolytica. The cysts can survive three to four
mouths outside the host’s body after desiccation.[8] The cysts cause infection by consuming
contaminated food and drinks like waste water. Sometimes insects and rodents carry the parasite
to cause infection in the food and drinks. Excystation happens once the cysts are ingested, and
travel to the large intestine. E. coli trophozoites can be distinguished by their wide and tapered
pseudopodia. They are often mistaken for E. histolytica due to their overlap in size. The cysts are
distinguished by noticing the eight nuclei found in the mature form.[10] To diagnosis for E. coli, a
stool sample is usually tested. This is the best method to check to see if the parasite is E. coli and
not E. histolytica. This usually involves checking the cysts for the size, shape, and the number of
nuclei. E. coli has cysts in size to 10 to 35 micrometers, the shape is irregular, oval with a shell-like
appearance that is more uniformed compared to E. histolytica, and has up to eight nuclei in the
cyst compared to the four nuclei of E. histolytica. To the untrained eye of by inexperienced
microbiologists, "tetranucleate cysts of Entamoeba coli can be mistaken for mature cysts of
Entamoeba histolytica" Often " a tetranucleate Entamoeba coli cyst is larger than a mature cyst of
Entamoeba histolytica, can be variable in shape, and has nuclear peripheral chromatin and
karyosome composed of irregular granules" in this matter of comparison. To make a diagnosis for
any Entamoeba species, usually a wet mount is created "by finding the characteristic cysts in an
iodine stained, formol-ether concentration method or by detecting the characteristic trophozoites
in a wet preparation or a permanent stained preparation" to see what they may look like.[11] Also,
these stains of trichrome can be used to mount the cysts of any Entamoeba spp.[12] Other tests
can be used to diagnosis for Entamoeba spp. These tests involve the use of laboratory methods.
Some of these laboratory tests include: the use of light microscopy, culture methods, isoenzyme
analysis, antibody detection tests, antigen detection tests, immunochromatographic assays, and
DNA-based diagnostic tests.[13] Some uses of microscopy also involve the use of transmission
electron microscopy and scanning electron microscopy. Usually, the cysts are freeze fractured to
insure that the samples are easier to look at to compare Entamoeba spp.[14] The DNA-based
diagnostic tests include the use of DNA extraction, PCR, microarrays, and typing methods.For
example, one DNA-based diagnostic test that is changing how Entamoeba spp. is being diagnosis
faster and more accurate is by using the "Reverse Line Hybridization Assay" test. This test main
purpose is to detect and different Entamoeba spp. in stool samples in order to find the causative
agent of amoebic dysentery, E. histolytica. This test involves the use of gene sequencing, and
seeing what different genomes each Entamoeba spp. has to help detect the deadly E. histolytica
Amoeba proteus: Amoeba proteus, también llamada Chaos diffluens es quizá la ameba más
conocida, pero ni mucho menos la más común. Su interior es como una vidriera móvil, con las
algas verdes, gotitas de grasa, cristales de proteínas, gránulos de almidón y plaquitas de leucina, se
mueven en un caleidoscopio asimétrico, todo dirigido por un solo núcleo, el de este
microorganismo gigante tan desconcertante, ya lo hemos dicho, como inofensivo. Ver video!!
La ameba es un organismo unicelular del género Amoeba que pertenece al filo Amoebozoa y al
reino protista. Los sistemas antiguos de clasificación incluían a las amebas entre los animales. La
ameba fue descubierta por el naturalista alemán August Johann Rösel von Rosenhof en 1757.1 Los
naturalistas se refirieron a la ameba como animal de Proteo, un dios griego que cambiaba de
forma y etimológicamente ameba procede del griego amoibè (αμοιβή), que significa cambio de
forma.2 De ahí procede el nombre científico de la especie Amoeba proteus.

Por extensión, se denomina ameba a cualquier miembro del filo Amoebozoa. La especie Polychaos
dubium (ameba dubia) es todavía mayor, pues mide más de un milímetro y es visible a simple
vista. Al menos seis especies de este grupo son parásitos del hombre. De éstas, la más importante
es Entamoeba histolytica, que causa la amebiasis o disentería amebiana. Las amebas de los
géneros Chaos y Pelomyxa pueden presentar cientos de núcleos, en contraste con Amoeba, que
sólo tiene uno. Las amebas de los géneros Arcella o Difflugia producen conchas o testas que les
sirven de protección.

Morfología de una ameba. En el sentido de las agujas del reloj: seudópodo, vacuola contráctil,
endoplasma, ectoplasma, membrana citoplasmática, núcleo y vacuola digestiva.

Las amebas tienen la estructura típica de una célula eucariota, presentando citoplasma, núcleo y
diversos orgánulos. El citoplasma se divide en una masa central granular denominada endoplasma
y una capa externa más clara llamada ectoplasma. Los elementos más reconocibles en la ameba
son el núcleo y la vacuola contráctil que emplea para mantener la presión osmótica. Esta vacuola
recibe el agua en exceso de la célula y periódicamente se une a la membrana citoplasmática para
expulsar el agua al exterior. Las vacuolas digestivas reciben el alimento una vez ingerido y lo
digieren.

Las amebas se desplazan extendiendo el citoplasma hacia afuera, formando prolongaciones

similares a tentáculos, conocidos como seudópodos o falsos pies. Los seudópodos se utilizan
también para envolver el alimento en un proceso conocido como fagocitosis. Cuando las
condiciones son desfavorables, las amebas pueden formar quistes, etapa de reposo en la que la
ameba se encierra en una bola segregando una pared protectora. El quiste puede sobrevivir a
condiciones ambientales mucho más duras que la ameba original. La célula permanece en estado
durmiente hasta que las condiciones se vuelven más favorables. Dentro del quiste la célula se
divide varias veces por mitosis, de forma que cuando el quiste se abre, salen varias amebas hijas.

Alimentación: La ameba es un organismo de nutrición heterótrofa pues se alimenta de toda clase

de plantas y animales microscópicos, de bacterias y de otras células. La formación de seudópodos
se produce como respuesta a estímulos químicos generados por microorganismos que constituyen
su alimento. Un ácido secretado en la vacuola descompone este alimento en sustancias químicas
solubles que son difundidas desde la cavidad al citoplasma. Por ende, es una digestión intracelular.
Este proceso es conocido como fagocitosis. El material de desecho y los restos no digeridos son
eliminados a través de las vacuolas del ectoplasma, el cual también absorbe oxígeno del medio
líquido en que se encuentra la ameba y elimina el dióxido de carbono originado en el
metabolismo. Se trata de una forma de respiración. Tras un período de crecimiento, la ameba se
reproduce por división en dos partes iguales.
Presenta un tamaño inferior a 20 μm. Carece de ciertos orgánulos como son las mitocondrias y el
aparato de Golgi. Únicamente tiene un hospedador (monoxeno), es cosmopolita y tiene dos
formas de vida en su ciclo vital:

Trofozoíto: presenta un tamaño en torno a 20 μm de longitud y 15 μm de ancho con una

morfología piriforme y una simetría bilateral. Proyectada en un plano se asemeja a una pera.
Posee 8 flagelos, 2 anteriores, 2 posteriores, 2 ventrales y 2 caudales, cuya función es la motilidad
celular. En la cara ventral presenta una estructura con forma de disco bilobulado, cuya función es
permitir la fijación del parásito a la superficie del epitelio intestinal. En la cara dorsal y
coincidiendo en posición con el disco bilobulado se sitúan dos núcleos ovalados con grandes
endosomas. A lo largo de la superficie ventral se disponen unos elementos denominados cuerpos
mediales, cuya función aún permanece desconocida. El trofozoito es la forma vegetativa que se
alimenta y se reproduce.

Quiste de Giardia con tinción de lugol Quiste: presenta un tamaño en torno a 15,4 μm de longitud
y 9,7 μm de ancho con una morfología ovalada. Posee 4 núcleos que siempre aparecen dispuestos
en alguno de los polos. No presenta flagelos aunque se pueden apreciar los axonemas flagelares
(restos de los flagelos) y los cuerpos mediales duplicados con respecto al trofozoito. La pared es
transparente y muy resistente tanto a factores físicos como químicos. El quiste es la forma
vegetativa infectante y de resistencia.
Alimentación por fagocitosis y pinocitosis del contenido intestinal a través de la superficie dorsal.

Reproducción por división binaria longitudinal. Se reproduce tan rápido que en poco tiempo
pueden formarse millones de parásitos. No presentan reproducción sexual.

Ciclo vital de Giardia lamblia. Giardia lamblia vive en forma de trofozoito en la luz del intestino
delgado (principalmente en el duodeno) adherido a las vellosidades intestinales por medio de los
discos bilobulados. Se alimenta y se reproduce hasta que el contenido intestinal inicia el proceso
de deshidratación, momento en el que comienza el enquistamiento del trofozoito. Pierde los
flagelos, adquiere una morfología ovalada, se rodea de una pared quística y madurez. Los quistes
expulsados junto a las heces ya son infectantes. Cuando dichos quistes son ingeridos por un nuevo
hospedador, llegan al duodeno, donde se disuelve la pared quística, dando así lugar a un individuo
tetranucleado que se divide inmediatamente en dos trofozoitos binucleados que se anclan al
epitelio intestinal, cerrando así su ciclo vital.

Patogenia La patología originada por G. lamblia se debe principalmente a los efectos que causan la
acción mecánica de adherirse y fijarse al epitelio intestinal. Dichos efectos producen una
alteración de las microvellosidades, que disminuyen su superficie de exposición al ser engrosadas,
y esto conlleva la aparición de diversas alteraciones fisiológicas más o menos graves, según el
mayor o menor deterioro del proceso de absorción. Cabe mencionar que la sustracción de
alimento producida por el parásito no parece ser relevante en la patogénesis. La patogenicidad
también se ve muy influenciada por el tipo de cepa y el estado inmunitario del hospedador y es
totalmente aeróbica.

Sintomatología Los síntomas producidos por una giardiasis pueden ser desde inexistentes hasta
presentar una sintomatología grave. En caso de que la infección curse con síntomas, estos
aparecen tras un período de incubación que dura en torno a 1-3 semanas, y consisten
principalmente en diarreas mucosas, sin restos de sangre y meteorismo, dolor abdominal y
anorexia (síntoma) . En los casos más severos se puede llegar a producir el síndrome de
malabsorción, debido a la destrucción de las células epiteliales del intestino delgado. Esto obliga a
un constante reciclaje de los epitelios con células inmaduras, que aún no son capaces de absorber
o digerir ciertas moléculas, lo que determina una malabsorción de lípidos, glúcidos y proteínas.
Está caracterizada por la aparición de esteatorrea (heces grasas y copiosas) y, posteriormente, de
deficiencias proteicas y vitamínicas (sobre todo vitaminas liposolubles). La duración de la fase
aguda de la infección es de unos 3 ó 4 días y va desapareciendo a medida que actúa el sistema
inmunitario del hospedador a través de los linfocitos T. En algunos individuos, principalmente
aquellos inmunodeficientes, la enfermedad puede hacerse crónica, pudiendo prolongarse los
síntomas durante años.
Paramecium caudatum

Pellicle - a membrane covering that protects the paramecium like skin

Cilia - hair like appendages that help the paramecium move food into the oral groove

Oral Groove - collects and directs food into the cell mouth

Cell Mouth - opening for food

Anal Pore - disposes of waste

Contractile Vacuole - contracts and forces extra water out of the cell

Radiating Canals - paths to the contractile vacuole

Cytoplasm - intercellular fluid needed to contain vital cell parts

Trichocyst - used for defense

Gullet - forms food vacuoles

Food Vacuole - storage pocket for food

Macronucleus - larger nucleus which performs normal cell functions

Micronucleus - smaller nucleus which is responsible for cell division.

The paramecium, genus of protozoa of the phylum Ciliophora, is often called slipper animalcules
because of their slipper-like shape. Paramecia are unicellular organisms usually less than 0.25 mm
(0.01 in) in length and covered with minute hair-like projections called cilia. Cilia are used in
locomotion and during feeding. When moving through the water, paramecia follow a spiral path
while rotating on the long axis. When a paramecium encounters an obstacle, it exhibits the so-
called avoidance reaction: It backs away at an angle and starts off in a new direction. Paramecia
feed mostly on bacteria, which are driven into the gullet by the cilia. Two contractile vacuoles
regulate osmotic pressure (see Osmosis) and also serve as excretory structures. A paramecium
has a large nucleus called a macronucleus, without which it cannot survive, and one or two small
nuclei called micronuclei, without which it cannot reproduce sexually. Reproduction is usually
asexual by transverse binary fission, occasionally sexual by conjugation, and rarely by endomixis, a
process involving total nuclear reorganization of individual organisms. Macronuclear DNA in
Paramecium has a very high gene density. The macronucleus can contain up to 800 copies of each
gene. Paramecia abound in freshwater ponds throughout the world; one species lives in marine
waters. They are easily cultivated in the laboratory by allowing vegetable matter to stand in water
for a few days. The common species Paramecium caudatum is widely used in research.
TRIPANOSOMAS
PLASMODIUM VIVAX
TRICHOMONAS VAGINALIS