05- Parasitology (Exam # 5) Flashcards
Define the following term.
Parasite
Organism that lives in or on and takes its nourishment from another organism.
Define the following term.
Obligate parasite
Microorganism or virus unable to live independently outside a living cell.
Define the following term.
Definitive host
Individual in which a parasite has its adult and/ or sexual reproductive stage.
Define the following term.
Intermediate host
Individual in which a parasite has its larval and or asexual reproductive stage.
Define the following term.
Accidental host
An organism that generally does not allow transmission to the definitive host, thereby preventing the parasite from completing its development.
Define the following term.
Reservoir host
Host who harbors a parasite that is also infective for humans
Define the following term.
Ectoparasite
Organisms that live on, rather than in, the human body; include fleas, lice, ticks, and mites.
Define the following term.
Endoparasite
A parasite, such as a tapeworm, that lives inside its host.
Define the following term.
Vector
Organism responsible for transmitting parasite from infected host to noninfected host.
Define the following term.
Periodicity
Recurring at a regular time period
Define the following term.
Nocturnal
Primary expression of the periodicity during the night
Define the following term.
Diurnal
Primary expression of the periodicity during daylight hours
Stool
Macroscopic examination of stools
- Consistency (liquid, soft, formed)
- The color of the stool specimen (brown, red, black, mucus)
- Intact worms or proglottids (tapeworm segments)
Stool
Microscopic examination of stools
1.Direct wet mount examination (stained with iodine and unstained) of fresh stool specimens. Unstained could show motility.
2.Concentration procedures on stools with wet mount examination of the concentrate
3.Preparation of permanently stained smears
Stool
List preservatives used for specimen stabilization
Stool
Describe the purpose of adding a fixative to the preservative
Trophozoites, the motile and reproductive form of some amebae, or eggs of some helminths may disintegrate if not preserved or examined within a short time.
What is the ratio of preservative to feces for optimal fixation?
3 parts preservative to 1 part feces
Stool
Discuss the number of and timing of specimen collection for optimal detection of intestinal parasites.
- Because many intestinal organisms are shed into the stool irregularly, a single stool specimen may be insufficient to detect an intestinal parasite.
- 58% to 72% of protozoa are detected with a single specimen.
- For optimal detection of intestinal parasites, a series of three stool specimens collected a day or two apart, within a 10-day period, has been recommended.
Stool
Discuss specimen rejection criteria
- Stool specimens should never be collected from bedpans or toilet bowls; such practice might contaminate the specimen with urine or water, resulting in the destruction of trophozoites or introduction of free-living protozoa.
- Stool specimens for parasites should be collected before a barium enema, certain procedures using dyes, or the start of antimicrobial therapy. Antimicrobials can reduce the number of organisms present.
- If the patient has undergone a barium enema, stool examination should be delayed for 7 to 10 days because barium obscures organisms when specimens are examined microscopically, even after concentration procedures.
- If a purged specimen is to be collected, it is recommended that a saline or phosphosoda purgative be used because mineral oil droplets interfere with identification of parasites, especially protozoan cysts, an infective dormant form resistant to environmental stress. The second or third specimen after the purge is more likely to contain trophozoites that inhabit the cecum.
Stool
Discuss the purpose of performing concentration techniques designed to enhance recovery of parasites
- Concentration techniques are designed to concentrate the parasites present into a small volume of fluid and remove as much debris as possible. Fresh or formalin-preserved stool specimens may be used. The concentrate sediment may then be examined unstained or stained with iodine.
- Protozoan cysts, helminth larvae, and helminth eggs can be detected using this method.
What protozoan stage does not survive the concentration procedure?
Protozoan trophozoites do not survive the procedure.
Name the 2 types of concentration techniques?
- Sedimentation and Flotation methods
- Both are based on the difference in specific gravity between the parasites and concentrating solution and are used to concentrate parasites into a small volume for easier detection.
Describe
Sedimentation method
- The organisms are concentrated in sediment at the bottom of the centrifuge tube.
- The sediment is used for wet mounts.
- The formalin– ethyl acetate sedimentation (FES) method is the standard sedimentation method.
Stool
Flotation method
- The organisms are suspended at the top of a high-density fluid.
- The fluid is extracted from the top for wet mounts.
- The zinc sulfate method is the usual flotation procedure.
Stool
Contrast the flotation method versus the sedimentation method for enhancing the detection of parasites.
Although the zinc sulfate flotation method yields less fecal debris in the finished preparation compared with the FES sedinentation method, the zinc sulfate causes operculated eggs to open or collapse. It also tends to distort protozoan cysts. When used, this procedure may miss infertile Ascaris lumbricoides eggs and Schistosoma spp. eggs. Because of their high density, these eggs sink to the bottom of the test tube. Overall, sedimentation methods concentrate a greater diversity of organisms, including cysts, larvae, and eggs.
What is included in an ova and parasite examination?
Discuss risk factors for acquiring a parasitic infection.
- Immunocompromised patients
- Travel to countries that have less than ideal sanitation and a large number of endemic parasite
- Ongoing climate change might affect the movement of insect vectors into new geographic areas, thus expanding the range of some diseases normally associated with tropical climates.
Discuss the rationale for identifying and reporting both presumed pathogenic and non-pathogenic parasites.
In the case of intestinal protozoa, it could indicate exposure to fecally contaminted food or water.
Urine, Vaginal, and Urethral Specimens
Describe the wet mount preparation for vaginal specimens
- Place one drop of saline-diluted fluid onto microscope slide. Coverslip and remove
excess fluid from the edges (if necessary) by gently blotting on edge against a paper
towel. Examine under 40x magnification with reduced light. - The trophozoites of Trichomonas, if present, move with a nervous, jerky motion.
They possess an undulating membrane, which extends only half the length of the
organism
Describe the collection of duodenal aspirates for parasite evaluation.
* For the detection of duodenal infections such as Giardia intestinalis or Strongyloides stercoralis.
- Collected using direct aspiration during endoscopy or with the “Beale String” technique AKA the duodenal capsule Entero-Test
- Performed in cases of suspected giardiasis or strongyloidiasis when clinical symptoms are suggestive of infection but repeated routine stool examination results are negative.
- In the Entero-Test, the patient swallows a gelatin capsule containing a weighted string. One end of the string is taped to the side of the patient’s mouth; the weighted end is carried into the upper small intestine. After about 4 hours, the string is brought up, and part of the mucus adhering to the surface is stripped off and examined on a wet mount for motile trophozoites. The remainder of the specimen is placed in a fixative for a permanently stained smear. Eggs of Fasciola hepatica and Clonorchis sinensis, as well as oocysts of Cryptosporidium and Cystoisospora belli, can also be recovered.
Discuss the importance of noting the macroscopic examination of unpreserved fecal specimens.
- Consistency may help determine the type of preservative to be used, indicate the forms of parasites expected to be present, or dictate the immediacy of examination.
- Fig. 28.1 shows the relationship between stool consistency and protozoan stage. Cysts (infective stage) are most likely to be found in formed stools and sometimes in soft stools, whereas liquid stools will most likely contain the trophozoite stage.
Discuss the purpose of wet mount preparations
- The direct wet mount of unpreserved fecal material is primarily used to detect the presence of motile protozoan trophozoites in a fresh liquid stool or from sigmoidoscopy material.
- Specimens should be examined within 1 hour and direct wet mount performed using saline & iodine solutions (note: iodine will kill trophozoites)
Explain the microscopy procedure for concentrated specimens
10x & entire slide
40x & at least 50% of slide
Discuss the purpose of preparing permanently stained smears
- Permanently stained smear preparations of all stool specimens should be made to detect and identify protozoan trophozoites and cysts.
- The characteristics needed for identification of the protozoa, including nuclear detail, size, and internal structures, are visible in a well-made and properly stained smear.
**In a well-stained trichrome smear, the cytoplasm of protozoan cysts and trophozoites stains blue-green, although Entamoeba coli often stains purple. Nuclear peripheral chromatin (deoxyribonucleic acid [DNA] and proteins on the edge of the nucleus), karyosome (mass of chromatin in the nucleus), chromatoidal bars (dark-staining cytoplasmic inclusions of chromatin), and RBCs stain dark red to purple. Eggs and larvae stain red; however, they are often distorted or destroyed in the staining process. Background debris and yeasts stain green. With an iron hematoxylin stain, the parasites stain gray to black, nuclear material stains black, and background material stains light blue to gray.
What are the most commonly used permanent stains?
- Iron hematoxylin and trichrome
- The stain of choice in most laboratories is the trichrome stain because results are somewhat less dependent on the technique and the procedure is less time-consuming.
How to calibrate an ocular micrometer
What is the size of the egg in um?
Contrast the use of enzyme immunoassays and microscopic exam for the detection of parasites.
Immunoassys are more sensitive and specific
What parasitic antigens can be detected by EIA?
Giardia, Cryptosporidium, Entamoeba histolytica
DFA (direct flourescent antibody) can be used for detecting which parasites?
Cryptosporidium and Giardia
Describe the purpose of performing an analysis for occult blood, white blood cells, lactoferrin and fecal fat
- Lactoferrin is a marker for fecal leukocytes (wbcs) and an indicator of intestinal inflammation.
- Leukocytes are found in stools in the presence of infection with bacteria that invade the colonic mucosa (i.e. Salmonella, Shigella, Yersinia and invasive E. coli).
- Other disorders that may be associated with fecal leukocytes are ulcerative colitis, Crohns disease and antibiotic associated colitis.
* Fecal leukocytes are usually absent in diarrhea secondary to parasites or viruses. - Useful for diagnosing fat malabsorption due to pancreatic or intestinal disorders
Recall appropriate therapy for the following parasites.
Giardia duodenalis (lamblia)
Metronidazole
Recall appropriate therapy for the following parasites.
Cryptosporidium parvum
Generally self limiting. Drink plenty of fluids to stay hydrated
Recall appropriate therapy for the following parasites.
Trichomonas vaginalis
Metronidazole
Recall appropriate therapy for the following parasites.
Enterobius vermicularis
Mebendazole
Explain the picture
- The Metro (metronidazole) runs over Entamoeba histolytica, Giardia lamblia, Trichomonas vaginalis, and the anaerobic organisms including Bacteroides fragilis, Clostridioides difficile and Gardnerella vaginalis.
- This drug is also called Flagyl (it’s trade name) because it kills the flagellated bugs, Giardia and Trichomonas
PROTOZOA
What are the general characteristics of Amoebas?
- Single celled
- Move by pseudopodia
- Trophozoite and cyst stages
- Asexual reproduction
Protozoa
Describe the intenstinal ameba life cycle.
The life cycle of amebae is relatively simple, with direct fecal-oral transmission in food or water via the cyst stage and no intermediate hosts. Humans ingest the infective cysts, which excyst in the intestinal tract, and the emerged trophozoites multiply by binary fission. Trophozoites colonize the cecal area.
Cyst
Infective form of a protozoan; characterized by the formation of a thick protective wall that is resistant to environmental factors.
Trophozoite
The motile and reproductive form of some protozoa
Karyosome
Spherical chromatin mass within the nucleus of protozoa.
Chromatoid body (bars)
RNA that has been condensed into a barlike structure within the cyst of some protozoan organisms.
Kinetoplast
Structure found in some blood and tissue protozoans composed of the granule, from which the flagellum or undulating membrane arises, and a mitochondrion.
Protozoa
Name the 2 intestinal pathogenic amoebas?
- Entamoeba histolytica
- Blastocystis hominis
What stage in the life cycle of intestinal amoeba is infective?
Cyst
What three intestinal amoeba have a centrally located karysome with even peripheral chromatin?
- Entamoeba histolytica/dispar
- Entamoeba hartmanni
Protozoa
Which form E. histolytica/E. dispar is depicted in the picture?
Trophozoites
Protozoa
These mature cysts have 4 nuclei that characteristically have centrally-located karyosomes and fine, uniformly distributed peripheral chromatin. Cysts usually measure 12 to 15 µm.
Entamoeba histolytica/Entamoeba dispar
Cyst of E. histolytica/E. dispar stained with trichrome. Three nuclei are visible in the focal plane (black arrows), and the cyst contains a chromatoid body with typically blunted ends (red arrow). The chromatoid body in this image is particularly well demonstrated.
Protozoa
Erythrophagocytosis (ingestion of red blood cells by the parasite) is classically associated with what organism?
Trophozoite with ingested erythrocytes stained with trichrome. The ingested erythrocytes appear as dark inclusions. The parasite above shows nuclei that have the typical small, centrally located karyosome, and thin, uniform peripheral chromatin.
E. histolytica
What host is the primary reservoir for E. histolytica?
Humans
Describe the clinical infections associated with E. histolytica.
- 90% are asyptomatic
- Most experience vague abdominal symptoms
- Severe infections develop liver or lung abscesses
Protozoa
This intestinal Amoeba is pathogenic
Blastocystis hominis
Protozoa
Identify this non-pathogenic intestinal Amoeba in its cyst form
Entamoeba hartmanni
Protozoa
Identify this non-pathogenic intestinal Amoeba in its cyst form
Entamoeba coli
What two intestinal amoeba lack peripheral chromatin?
- Endolimax nana
- Iodamoeba butschlii
Protozoa
Identify this non-pathogenic intestinal Amoeba in its cyst form
Endolimax nana
Protozoa
Identify this non-pathogenic intestinal Amoeba in its cyst form
Iodamoeba buetschlii
Protozoa
Name 3 Nonintestinal (free living) amoeba?
- Acanthamoeba spp
- Naegleria fowleri
- Balamuthia mandrillaris
What stage in the life cycle of free-living pathogenic amoeba is infective?
Trophozoite
Protozoa
Identify the cyst form of this nonintestinal (free living) pathogenic amoeba
Acanthamoeba spp
Protozoa
Describe the life cycle of Acanthamoeba
Route of infection is direct inoculation
Acanthamoeba spp. have been found in soil; fresh, brackish, and sea water; sewage; swimming pools; contact lens equipment; medicinal pools; dental treatment units; dialysis machines; heating, ventilating, and air conditioning systems; mammalian cell cultures; vegetables; human nostrils and throats; and human and animal brain, skin, and lung tissues. Unlike N. fowleri, Acanthamoeba has only two stages, cysts (1) and trophozoites (2), in its life cycle. No flagellated stage exists as part of the life cycle. The trophozoites replicate by mitosis (nuclear membrane does not remain intact) (3). The trophozoites are the infective forms, although both cysts and trophozoites gain entry into the body (4) through various means. Entry can occur through the eye (5), the nasal passages to the lower respiratory tract (6), or ulcerated or broken skin (7). When Acanthamoeba spp. enters the eye it can cause severe keratitis in otherwise healthy individuals, particularly contact lens users (8). When it enters the respiratory system or through the skin, it can invade the central nervous system by hematogenous dissemination causing granulomatous amebic encephalitis (GAE) (9) or disseminated disease (10), or skin lesions (11) in individuals with compromised immune systems. Acanthamoeba spp. cysts and trophozoites are found in tissue.
Protozoa
Identify the cyst form of this nonintestinal (free living) pathogenic amoeba
Balamuthia mandrillaris
Protozoa
Describe the life cycle of Balamuthia mandrillaris
Balamuthia mandrillaris has only recently been isolated from the environment and has also been isolated from autopsy specimens of infected humans and animals. B. mandrillaris has only two stages, cysts (1) and trophozoites (2), in its life cycle. No flagellated stage exists as part of the life cycle. The trophozoites replicate by mitosis (3). The trophozoites are the infective forms, although both cysts and trophozoites gain entry into the body (4) through various means. Entry can occur through the nasal passages to the lower respiratory tract (5), or ulcerated or broken skin (6). When B. mandrillaris enters the respiratory system or through the skin, it can invade the central nervous system by hematogenous dissemination causing granulomatous amebic encephalitis (GAE) (7), or disseminated disease (8), or skin lesions (9) in individuals who are immune competent as well as those with compromised immune systems. B. mandrillaris cysts and trophozoites are found in tissue.
Protozoa
Identify this nonintestinal (free living) pathogenic amoeba
Naegleria fowleri
Describe the causative parasitic agent and the disease syndrome.
Primary amebic meningoencephalitis (PAM)
Naegleria fowleri
- The trophozoite enters the nasal cavity through inhalation of contaminated water or soil. The amebic form colonizes the nasal cavity, invades the nasal mucosa, attaches to olfactory nerves, penetrates the cribriform plate, moves along the olfactory nerve to the olfactory bulb, and moves into the arachnoid space. From there, it is free to spread throughout the CNS.
- Clinically, the disease cannot be distinguished from bacterial meningitis.
- The organism multiplies in brain tissue, and within 2 to 4 days, the patient can experience drowsiness, confusion, and seizures, and progress to coma. The disease usually is fatal within 1 week of the appearance of clinical symptoms.
Protozoa
Which free living amoeba causes GAE (granulomatous amebic encephalitis)?
Both Acanthamoeba & Balamuthia mandrillaris
Protozoa
Amebic keratitis in contact lens wearers is caused by which free living amoeba?
Acanthamoeba spp
Ussually due to contaminated lens care solution
Protozoa
Cultures can be performed on a lawn of bacteria (E. coli) for which two free living amoebas?
Both Acanthamoeba & Balamuthia mandrillaris
SPOROZOA
Protozoa
What class of protozoa reproduce both sexually and assexually in thier life cycle and are nonmotile?
Sporozoa
What stain is used to identify coccidia?
Modified Acid-fast
Stains the oocyst
What 3 organisms make up the coccidia?
- Cryptosporidium parvum
- Cystoisospora belli
- Cyclospora cayetanensis
Oocyst
The encysted form of the fertilized macrogamete, or zygote, in coccidian Sporozoea in which sporogonic multiplication occurs; results in the formation of sporozoites, infectious agents for the next stage of the sporozoan life cycle.