University of Guyana

Faculty of Natural Sciences

Department of Biology

BIO3219: Parasitology

Lecturers: Prof. Ansari & Mr. Lakenarine

Lab Instructor: Ms. Renea Douglas

Laboratory Exercise # 5
Blood Parasites

Ronaldo Binda
1036568
Aim: To examine blood samples for blood parasites.

Introduction:

Blood samples can be very useful in diagnosing parasitic infections because many parasites can

be detected in the blood of infected individuals. Here are some examples of how blood samples

can be used to diagnose parasitic infections: Blood smears - are prepared by placing a drop of

blood on a microscope slide and staining it with a special dye which allows the examiner to look

for parasites, such as the malaria parasite or trypanosomes, which can be seen under a

microscope. Serology - involves testing for the presence of antibodies produced by the body in

response to an infection. Blood samples can be tested for the presence of specific antibodies

against parasitic infections, such as toxoplasmosis or schistosomiasis. Polymerase chain reaction

(PCR) - a laboratory technique that can detect the DNA of parasites in blood samples. This can

be useful for diagnosing parasitic infections that are difficult to detect using other methods, such

as leishmaniasis or babesiosis. Blood culture - involves incubating a blood sample to see if any

parasites grow. This can be useful for diagnosing certain parasitic infections, such as babesiosis

or trypanosomiasis (Merck Manual Professional Version, n.d.). These blood samples can be very

useful in diagnosing parasitic infections, and different tests can be used to detect different types

of parasites, additionally, these tests are determined based on symptoms and medical history of a

person (UpToDate, n.d.).

Method:

Finger stick:

1. End of finger was cleaned with disinfected solution.

2. Fingertip was wiped with sterile material (remaining disinfectant was removed since it

would interfere with diagnostic process).

3. Fingertip was pierced with sterile lancet.

4. Blood was allowed to flow freely, finger was not squeezed.

Thick Blood Smear Preparation:

1. A small drop of blood was placed in the center of a pre-cleaned labeled slide.

2. The corner of another slide or and applicator stick was used to spread the drop into a

circular pattern until it was the size of a dime (1.5cm2).

3. A thick smear of proper density was one in which, when placed (wet) over newsprint, had

allowed a person to barely read the words.

4. The slides were laid flat and the smear was allowed to dry thoroughly for a minimum of

thirty minutes (protected from dust and insects). Insufficiently dried smears or smears too

thick could have detached from the slide during staining.

5. Staining was proceeded with as outlined below.

Thin Blood Smear Preparation:

1. A clean and unscratched microscope slide was chosen.

2. The slide were washed with alcohol to remove any residual oil or dust.

3. The slide were labeled with pencil on the frosted end, with the sample number and

details.
 4. The slides label was checked with the sample bottle- they should have been the same.

5. One small drop of fresh blood was placed on one end of the slide.

6. The end of another slide was used at an angle of approximately 30° to the first, the

second slide was allowed to be moved along the first slide to spread the blood drop into

one continuous thin film.

7. The smear was air dried before staining.

Staining of blood smears

1. Blood film were fixed in absolute methanol (acetone-free) for 30 seconds. Thin smear

was dabbed with methanol-soaked cotton ball.

2. Slide was allowed to air dry (blood cells will now not lyse during next step).

3. Slides were immersed in Giemsa solution for 30 minutes.

4. "Floating" excess stain was gently rinsed off slide with buffered water; care was taken

not to wash the blood smear away.

5. Stain was rinsed until no more was seen in solution.

6. Smear was dried side down, making sure that smear did not touch the slide rack or other

surface used for drying. Slides were briefly dipped in water.

7. Slides were drained in vertical position and was allowed to air dry.

8. Slide then ready for microscopic examination.

Results:
Discussion:

1. Compare your results to that of five other classmates. Did they find any parasites? What

are the symptoms and treatments for these infections?

The blood smear results of the researcher showed to have no sign of blood parasites present,

however, only 3 blood parasites were found throughout the entirety of the class. These include

the assumed presences of Wucheria bancrofti in 2 classmates’ blood and Trypanosoma cruzi in

the epimastigote stage in another classmate. Wuchereria bancrofti is a parasitic filarial worm that

causes lymphatic filariasis, a tropical disease characterized by chronic swelling and inflammation

of the lymphatic system. Symptoms include: Swelling of the arms, legs, breasts, or genitals

(lymphedema), thickening and hardening of the skin in affected areas (elephantiasis), fever,

chills, and fatigue, pain and discomfort in the affected limbs, and recurrent episodes of

inflammation (acute attacks) that cause fever, severe pain, and swelling (WHO, 2022). Treatment

includes: Antiparasitic medications such as diethylcarbamazine (DEC) or ivermectin are used to

kill the microfilariae and adult worms, respectively, in some cases, surgery may be needed to

remove excess fluid or tissue in the affected areas, persons with lymphedema may need to wear

compression garments and practice good skin hygiene to prevent infections and further swelling,

and preventive measures such as avoiding mosquito bites and mass drug administration programs

may also be employed to control the spread of the disease (CDC, 2018). Trypanosoma cruzi is a

parasitic protozoan that causes Chagas disease, symptoms include: acute phase which causes

fever, fatigue, body aches, headache, rash, and swelling at the site of infection (e.g., around the

eye), and the chronic phase which causes heart rhythm abnormalities, difficulty swallowing, and

abdominal pain. In severe cases, Chagas disease can lead to heart failure, digestive disorders, and

sudden death. Treatment includes: antiparasitic medications such as nifurtimox or benznidazole

are used to kill the parasite and treat the infection. In the acute phase, antiparasitic treatment is

highly effective, and most patients can recover completely. In the chronic phase, the goal of

treatment is to manage the symptoms and prevent complications. Treatment is less effective in

this phase, and some patients may require surgery or other medical interventions. Regular

medical monitoring and follow-up are important to assess the progression of the disease and the

response to treatment (Medline Plus, 2022).

2. What parasites other than those observed can be diagnosed by using blood smears.

Since blood smears are a useful diagnostic tool for detecting several parasites that circulate in the

blood. Some other examples of blood parasites that can be detected from blood smears include:

Malaria parasites- caused by several species of Plasmodium parasites, and these parasites can be

seen in a blood smear under a microscope, Babesia parasites - are transmitted by ticks and cause

babesiosis. They can infect red blood cells and can be detected in a blood smear (CDC, 2018),

and Leishmania parasites- can cause leishmaniasis, a disease that can affect the skin, mucous

membranes, or internal organs. They can be detected in a blood smear, but the sensitivity of the

test may be low (WHO, 2022).

3. Limitations/ challenges encountered.

 Identification of blood parasites- The interpretation of blood smears requires expertise

and training. A skilled technician or medical professional is needed to prepare and

examine the smear under a microscope.

 False negatives- these results occur due to factors such as poor quality smears, inadequate

staining, or incorrect interpretation of the results, or presence of artefacts in the blood

which looks similar to blood parasites.

  Sensitivity- blood smears can vary depending on the type of parasite and the stage of

infection. In some cases, the number of parasites in the blood may be too low to be

detected by a blood smear.

Conclusion:

In conclusion, blood samples are a useful tool in diagnosing parasitic infections. Blood smears

can be used to detect parasites such as malaria, trypanosomes, microfilariae, babesia, and

leishmania. Serology, PCR, and blood culture are other diagnostic methods that can be used to

detect parasitic infections. Wuchereria bancrofti causes Lymphatic Filariasis and can lead to

swelling and Elephantiasis, and Trypanosoma cruzi causes Chagas disease and can lead to heart

and digestive problems. Antiparasitic medications and surgery may be used to treat these

infections. However, it is important to seek medical attention if a person suspects that they have

a parasitic infection as early diagnosis and treatment can prevent complications.

References:

American Society for Microbiology, 2017. Laboratory Diagnosis of Parasites from the

Gastrointestinal Tract. https://jcm.asm.org/content/55/10/3199

Centers for Disease Control and Prevention (2018). Parasites- Lymphatic Filariasis - Treatment.

https://www.cdc.gov/parasites/lymphaticfilariasis/treatment.html

Centers for Disease Control and Prevention (2018). Parasites- Babesiosis – Diagnosis

https://www.cdc.gov/parasites/babesiosis/diagnosis.html

Centers for Disease Control and Prevention (2018). Parasites- Malaria: Diagnosis and Treatment.

https://www.cdc.gov/malaria/diagnosis_treatment/diagnostic_tools.html

Medline Plus, 2022. Chagas Disease. https://medlineplus.gov/chagasdisease.html

Merck Manual Professional Version (n.d.). Diagnosis of Parasitic Infections.

https://www.merckmanuals.com/professional/infectious-diseases/intestinal-protozoa/diagnosis-

of-parasitic-infections

UpToDate, n.d. Laboratory diagnosis of parasitic infections.

https://www.uptodate.com/contents/laboratory-diagnosis-of-parasitic-infections

World Health Organization, n.d. Diagnostic methods for the detection of parasitic infections.

https://www.who.int/tdr/publications/documents/diagnostic-methods.pdf

World Health Organization, 2022. Lymphatic Filariasis. https://www.who.int/news-room/fact-

sheets/detail/lymphatic-filariasis

World Health Organization, 2022. Chagas disease (also known as American trypanosomiasis).

https://www.who.int/news-room/fact-sheets/detail/chagas-disease-(american-trypanosomiasis)