Malaria Case Study

Arln Toro Rafael Tosado Cole Benton

Malaria Case Studies

Potential Courses
Gen. Biology Intro. Ecology Gen. Microbiology Parasitology Gen. Biology Microbiology Parasitology Bioinformatics Genetics Molec. Biology Evolution Gen. Biology Parasitology Ecology Evolution Population Biology

Study Aspect Case1 Malaria prophylaxis and control

Case Scenario Tempting family vacation package but no time for malaria prophylaxis

Resources/Activity Maps Basic concepts Malaria Life Cycle Travelers info. WHO, CDC, NIH Bol Wrkbench Case it NCBI Plasmo DB CDC, WHO, NIAID HIPRE My World GIS Transgenic mosquitoes

Case2 Parasite-drug resistance and diagnosis challenges Case3 Ecology of the mosquito vector

Aunt Milagros died of cloroquineresistant malaria in 1982. Students collecting mosquitoes find Anopheles in a nonendemic area for malaria.

Student products:

develop ilustrated materials to use w/ deaf people, five min. comercials on malaria prophylaxis, etc.

As he munched yet another piece of Christmas rice pudding Sebastin Romn glanced about the gaily decorated room at his family. His brothers children were busily, excitedly comparing the presents they had pulled from underneath their beds earlier in this celebratory Three Kings Day. Family, food, music, and the smell of ginger and cinnamon smoldering on the table and sending small fragrant wisps throughout the house made everyone happy. Not everyone, Sebastin thought as he noticed his father staring out the kitchen window, a deep sadness on his face. What is it? Why arent you happy? Sebastin inquired, approaching his father and pulling back the curtain so he could look out at what his father saw.
Im OK, he said. I was just recalling that when you were just a small nio, twenty years ago on this day, we lost Aunt Milagros. Its the ginger smell, I guess, that does it, that makes me remember that sad, sad day. I loved my sister and I miss her deeply.

You never talked about what happened to her, Sebastin said, putting his arm around his fathers shoulder. What did happen?
Mr. Romn pulled his son toward the table where they would soon enjoy the Feast of Epiphany on this, the last day of their Christmas. They sat down. In a word, malaria. Sergio Romn told his son. Malaria. Romn repeated. It could have been anything. It happened, it took her from us and that was all that mattered. Still, I never really understood that disease. Talking seemed to brighten Mr. Romn. He enjoyed talking to this son who had flown home to their family from his university in the United States to share the holidays. Sebastin was a man now, a man a father could talk with.

 We hadnt had malaria here in Puerto Rico for over twenty years when your Aunt Milagros died of it. He went on, She was using her teaching skills to educate children in the Amazon Choc forest. Evidently she became infected on that trip and came home with the parasite already in her body. Sebastins mother Maria sat down beside her husband. She knew precisely what he would be talking about so seriously on this day, but she pretended ignorance. So, what are we discussing here? Aunt Milagros, Sebastin answered

I should have known. Your father always, as I do, remembers your aunts passing on this day. Was he finally discussing it with you?

Yes, he told me about her missionary work in Colombia. Sergio continued. She took her antimalarial drug Chloroquine as a preventative, but it didnt help. I just dont understand.

When she returned to Puerto Rico, she told us that she had had a fever the day before. The second day home here in Barranquitas she had it again. Then she had it again in two days after that. Your aunt insisted that the fever cycles were not, absolutely not, malaria because: I took the drugs as required by the regimen exactly as I was supposed to. Maria continued her husbands story.
I believed her. I still do. Sergio said.

There was a brief silence, then Maria, her hands moving across her chest, said what she didnt want Sergio to have to say again. She died a few days later.

Etiology and Epidemiology of Malaria

 Malaria is a parasitic disease caused by a protozoan that replicates within red blood the cells (RBCs). The parasite is transmitted by a mosquito. Endemic malaria happens mainly in tropical climates where the mosquito is normally found. Countries at risk are those where mosquito transmission is possible but not happening currently. Seasonal outbrakes (mainly in summer months) have been reported in places such as Switzerland, England and New Jersey.

www.who.org/en/

www.who.org/en/

 More than 100,000 people of all ages die of malaria each year according to the World Health Organization (WHO). Malaria is the second most devastating infectious disease in the world after AIDS. In 1980 malaria was endemic to 11 countries in the world. Currently, malaria transmission by mosquitoes happens in 107 countries according to the WHO.

www.who.org/en/

Level 1 Questions
Biology of Malaria
1. Which are the etiologic agents of malaria?

2. Which species of the mosquitoe is responsible for transmitting the disease?

3. Describe the parasites life cycle.

Online Resources on Malaria

CDC Malaria

Medline Plus NIH

Medline Plus Tutorial NIH

Malaria Triad: Genetics & Genomics

WHO/TDR Malaria Database

Challenfes for the Diagnosis of Malaria

Red blood cells (RBCs) infected with Plasmodium falciparum become sticky and tend to adhere to the inner wall of blood vessels. Sequestred RBCs may not be evident in a smear of peripheral blood at the early onset of infection when the ammount of infected RBCs is low (low parasitemia).

Animated movie: sequetred Rbcs infected With P. Falciparum

(www.whei.edu.au)

Click here view videos Penetrate and Burst

 Microscopic identification of malaria in peripheral blood requires an experienced microscopist, especially at low parasitemias. Both thick and thin blood smears should be performed in microscope slides to assess parasitemia.

Thin smear preparation video demonstration: (vet.upenn.edu)

Side of fingertip

Click here

Thick and Thin Blood Smears

Quic kTime an d a TIFF (Unc omp resse d) decompres sor are neede d to s ee this picture.

Thick blood smear

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deep view of blood

Thin blood Smear

Sinle-layer of RBCs
adapted from www.cdc.gov

Level 2 Questions
Malaria Diagnosis
1. Establish a relationship between the plasmodium life cycle and the periodicity of the fever episodes. 2. When would be the best time to take a sample of peripheral blood for a smear?

3. Do antibody-screening tests detect DNA mutations in in Drug-resistant strains? 4. Design a PCR-based method for the identification of chloroquine resistant strains of P. falciparum?

Online Diagnostic Resource

DPDx - CDC - Division of Parasitic Diseases
Complete Information on diagnostic methods for parasitic disease

Click here And search on: Malaria and Diagnostic methods

Malaria Evolutioln and Bioinformatics

 With the use of the insecticide dichlorodyphenyl-trichloroethane (DDT) and the drug chloroquine, malaria was eradicated from most urban areas around the world in the 1950s. Since the late 1970s the parasite began to develop resistance to chloroquine and the mosquito developed resistance to DTT.

www.cdc.gov

 There are different mechanisms of resistance related to different antimalarial drugs. Resistance to chloroquine and mefloquine in P. falciparum arose simultaneously in the early 1980s in South America and South east Asia. Mutations in the pfmdr gene have been associated wit multi-drug resistance. This gene encodes a trans membranefflux pump.

Malaria basics
CDC Malaria MedlinePlus NIH MedlinePlus Tutorial NIH

Malaria Triad: Genetics & Genomics

WHO/TDR Malaria Database

Level 3 Questions
Using bioinformatics tools, find the protein sequence with higher similarity to the Plasmodium falciparum mdr. Based on the sequence alignments of the mdr protein find potential differences in the structure responsible for drug resistance. P. falciparum is more virulent than the other species that cause malaria in humans. Can this be explained in evolutionary terms? Compare the mdr protein sequence from different Plasmodium species.

Retrieving protein sequences from NCBI Protein Data Bank

Retrieving protein sequences

Click on the NCBI link on the right side. Search for Plasmodium falciparum mdr Select Protein: sequence database Establish limits (ref seq) Change the display mode to FASTA

NCBI

Write your keyword Plasmodium Click go.

Click on Protein: sequence database

You will obtain an incredible number of matches. Lets type Plasmodium falciparum.

Notice the number of hits is lower. Lets try adding mdr (multidrug resistance) and click go.

Now you got only 19 hits. Click on limits to look for curated data

Select from this pull down menu RefSeq. Click go

This is your accession number, write it down. You will need this number if you need to retrieve this number. Click on the accession number

The first part of the record consist of general protein information. This number is the accession number of GenBank. If you click on it you will be directed to the gene sequence.

When you scroll down the record you will find information related to the protein function.

The last part of your record is the protein sequence. Now, lets compare this protein with other known proteins.

Click on Blink

Select from this pull out menu FASTA.

Click on best hits

To view the structure of

Select sort by taxonomy proximity Observe that your list order is different

Click select GI list

Select the sequences the first two sequences.

All the sequences that you selected will appear in this format

Select from this pull down menu send to file Save it

In this part you are going to work with Biology Workbench

Biology Workbench
If you have no experience with Biology workbench click on the Tutorial, complete the tutorial before you start with this unit. If you just want a quick review you may proceed to the next part.

Biology Workbench Tutorial

 Biology Workbench its free but you have to open an account. Select the Biology Workbench link.

Biology Workbench

Register to create an account in Biology Workbench.

Enter the Biology Workbench.

Scroll down to the button of this page.

Select the session tools.

Select start new session from the pull out menu.

Write down the name of the session. Click on the Start Session button

Select Protein Tools

From the pull down menu select add new protein sequence. Click on run button.

Browse your sequences Click to upload the file

Click on save

Click on save

Click on both sequences

On the pull down menu select CLUSTALW Click on Run

Accept all the parameters by clicking on submit button

Scroll until you see the sequence. Compare your sequences. Check on the key colored legend. Compare in terms of identity

Click on import alignments

First, select your alignment Second, select on the pull down menu TEXTSHADE Third, Click on Run

Click on submit button

Your results will be in this format which is easier to analyze.