MALARIA

Dr.Saroj Bapna Department of Zoonosis

Malaria is an infectious disease caused by a protozoan parasite Plasmodium. There are four identified species of this parasite causing human malaria.

1. 2. 3. 4.

Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale Plasmodium malariae.

MALARIA FACTS
Malartia has caused immense suffering to mankind. There are 500 million people suffering from malaria and approximately 5 million people die every year. Every 30 seconds, a child dies of malaria in Africa, almost 3,000 children each day. Up to 23 percent of African infants are born with the malaria parasite.

TRANSMISSION
Malaria is transmitted by the female Anopheles mosquito.The parasite undergo asexual division in human host and sexual multiplication in mosquito host . The asexual cycle completes in two phases Pre-erythrocytic phase in liver cells Erythrocytic phase in blood cells
Anopheles

Pre-erythrocytic schizogony
The infective form of the parasite is called sporozoites. Once in the body after the mosquito bite, these sporozoites reach the liver parenchyma cells in 3040 minutes and start developing. Each sporozoite multiplies into 20-30000 merozoites, ruptures the liver cell and is liberated into the blood stream where it infects red blood cells. This phase of development within the liver is termed as Pre-erythrocytic schizogony. Since only a few hepatocytes are affected, this phase does not produce any symptoms.

Erythrocytic schizogony

Merozoites released from the liver schizonts infect red blood cells. The ring stage trophozoites mature into schizonts, releasing merozoites which infect fresh red cells. Some parasites differentiate into sexual erythrocytic stages (gametocytes). Blood stage parasites are responsible for the clinical manifestations of the disease.

Malaria parasite in mosquito host

The male and female gametocytes, are ingested by Anopheles mosquito during a blood meal. fertilization of male and female gametocytes produce zygote. The zygotes in turn become motile ookinetes which burrow into the midgut wall of the mosquito where they develop into oocysts . The oocysts grow, rupture, and release sporozoites , which make their way to the mosquito's salivary glands and are injected to a new host during the next blood meal.

PREVENTION AND CONTROL

Control of malaria involves control of three living beings and their environment. Man (the host) Plasmodia (the agent) Female Anopheles mosquito (the vector) Insecticide-treated bed nets currently are one of the most effective weapons in the war on malaria because they are physical barriers, rather than a biological or chemical obstacle that mosquitoes can mutate to overcome.

Action of Chloroquine
The major action of chloroquine is to inhibit the formation of hemozoin from the heme released by the digestion of hemoglobin. Chloroquine accumulates in the food vacuole of the parasite and bind heme. This binding may prevent the heme being incorporated in to hemozoin crystal. As a result heme is accumulated in large quantity and leads to parasite death. Chloroquine resistance is due to a decreased accumulation of chloroquine in the food vacuole.

Is a malaria vaccine feasible ?

Immunity to malaria
Several lines of evidence suggest that protective immunity to malaria is possible
People living in endemic areas who are repeatedly exposed to malaria, develop a naturally acquired clinical immunity that controls the parasites in their blood, and that also protects them from severe disease Immunization with irradiated sporozoites has shown partial or complete protection in mice, monkeys and humans when they are subsequently exposed to malaria Experiments that demonstrate the effectiveness of passive transfer of immunity in human volunteers.

WHY DEVELOPING A MALARIA VACCINE

IS DIFFICULT ?

1.

There are 4 species of malaria parasite and each species composed of a number of genetically different strains. Each strain require different vaccine.

2. During the course of malaria infection the human host confronts four distinct life stages. Each of these stages presents different antigens (targets) to the immune system.
3. The parasite has evolved a series of strategies that allow it to confuse, hide, and misdirect the human immune system.

Strategies for the development of Malaria vaccine

1. Pre-erythrocitic vaccine: Vaccine that induce an immune response that would prevent the sporozoite from entering the hepatocytes or destroy it once it invaded the hepatocytes (Disease preventing vaccine). Asexual blood stage vaccine: Blocking merozoite invasion of red cells and inhibiting development of schizonts (antidisease or asexual stage). Transmission-blocking Vaccine: would arrest the development of the parasite in the mosquito. The antigamete vaccine would not protect illness or infection in the vaccinated individual but would help in preventing the spread of disease within a population.

2.

3.

Advances in malaria research

The complete Genome of all the three organism involved in the transmission of malaria.

Boosting mosquito immunity to malaria.

I. Genetic modification in mosquito to make them poor vector for the parasite.

II. Malaria resistant mosquito by genetically modifying bacteria that occur naturally in mosquitoes. Spit test.

Malaria Genome
The complete genome of all the three organism involved in transmission of malariathe parasite Plasmodium falciparum, mosquito vector Anopheles and the human host,has provided new tools for malaria research. the genes and the pathway involved in the transmission, pathogenicity and drug response of malaria and also opens doors for new drugs and vaccines development.

Genetic modification in mosquito

Transient gene silencing technology to block the

development
mosquito.

of

the

parasite

in

Anopheles

European Molecular Biologist discovered a pair of mosquito genes when one gene, called CTL4, is

inactivated, the mosquitoes destroy up to 97% of

the parasites developing inside their bodies. When the other, called LRIM1, is removed, it has the

opposite effect: the parasites multiply readily.

Genetic modification in bacteria

Researchers at Johns Hopkins Malaria Research Institute identified Bacteria in the gut of the Anopheles mosquito inhibit infection of the insect with Plasmodium falciparum, removing these bacteria, or microbial flora, with antibiotics made the mosquitoes more susceptible to Plasmodium infection because of a lack of immune defence.

Genetic modification in bacteria..

The benefit of cultivating (GM) genetically modified bacteria is that it is much easier to grow

large number of bacteria than to grow large number of mosquitoes. To spread these GM bacteria in the wild
population by creating artificial refuges (feed) for mosquitoes containing cotton balls impregnated with sugar and GM bacteria.

Flightless Mosquitoes
A new strain of genetically altered male mosquitoes passes their genes to females making the next generation unable to fly. This alteration disrupts wing muscle development in female. Males ability to fly is unaffected, and they show no ill effect from carrying the gene.

Spit Test
Genetic and antigen tests to find malaria infection
in saliva. An easy and painless saliva test would help

screening

more

people

who

don't

present

symptoms(asymptomatic carriers) of the illness.

Global warming and malaria

Mosquitoes are cold-blooded, which means their body temperature and their ability to nurture the Plasmodium parasite is affected by the temperature of their surroundings. With increasing temperature due to global climate change, potential new risk areas will increase, where lower temperatures presently limit the mosquito populations and the parasite's ability to develop in them.

Are sterile mosquitoes the answer to malaria elimination?

Sterile insect technology (SIT), involves generation of male mosquitoes, which are incapable of producing offsprings despite being sexually active.
This leads to an increase reduction in the population over time, in contrast to insecticides, which kill a certain fraction of the insect population.

Malaria culture
Malaria culture is the method to grow malaria parasites continuously in an invitro environment. Infected red blood cells are incubated in a culture bottle at 37 C together with a nutrient medium RPMI 1640 and serum or serum substitute Albumax. A special feature of the incubation is the special gas mixture mostly nitrogen (93% nitrogen, 4% CO2 and 3% O2) allowing the parasite to grow at 37 C in an incubator.

Malaria culture An alternative to gassing the culture with the

exact gas mixture is the use of candle jar. The candle jar is an airtight container in which the
culture and a lit candle are placed. The burning candle consumes some of the Oxygen and produces

CO2 which

acts

as

a fire extinguisher

at

approximately 5% Co2 concentration the candle

stops burning. The number of parasites increases by 2 to 3% every 48 hours (= one cycle). The

parasitemia can be determined via blood film

Candle Jar

Mosquito- the vector

Mosquitoes are insect vectors responsible for the transmission of parasitic and viral infections to millions of people worldwide, with substantial morbidity and mortality. An understanding of mosquito classification, distinguishing features, and the insect life cycle is important for planning disease surveillance as well as for designing and implementing effective measures for disease control and prevention.

Adult female mosquitoes

Anopheles Aedes

Culex

Life cycle of mosquito

There are four distinct stages in the life cycle of mosquito- egg, larva, pupa and adult.

Life cycle of Aedes mosquito

Mosquito Life Cycle

Egg

1. Aedes mosquitoes lay their eggs singly just above the water line in areas that are periodically wet. The egg can remain dormant until they are flooded or conditions are favorable for hatching. Aedes prefer to lay their eggs in artificial water filled container or tree holes. 2. Culex species prefer polluted standing water with large amount of organic material. Eggs are laid on the water surface in rafts, 100 or more eggs. 3. Anopheles mosquitoes breed in permanent bodies of fresh water. Eggs supported by flots on each side, are laid singly on the surface of water. 4. Eggs laid on water surface hatch in one to three days

Eggs
ANOPHELES

AEDES

CULEX

Larva

The egg hatch into first stage larva. There are four larval stages.

Most larvae (Aedes and Culex) have siphon tubes for breathing and hang upside down from the water surface.

Anopheles larvae do not have a siphon and lie horizontally at the surface of the water.
The larva feed on micro-organisms and organic matter in the water. On the fourth molt the larva changes into a pupa

Mosquito larva

Larvae

Pupa

The fourth stage larvae transform into pupae.

The pupal stage is a resting, non-feeding stage. Inside the pupal skin the development of adult mosquito takes place.

Within two to three days the mosquito emerges as an adult.

EMERGING ADULT

Adult

Adult males and females feed on plant sugars and nectar for energy. The females also feed on blood to provide nutrients for the developing eggs. Anopheline adults rest with an angle to the surface, whereas Aedes and Culex species keep their bodies parallel to the surface, which makes them easy to identify when sitting on the skin Aedes mosquitoes are day-biters whereas Culex bites at dusk and after dark.