Avian Malaria Introduction Malaria is a protozoal disease caused by members of the genus Plasmodium.

Infections may occur in both humans and animals. Approximately 350-500 million people are infected annually, with 1.3 million deaths. It is endemic to parts of Asia, Africa, Central and South America, and certain Caribbean islands. There are four species of Plasmodium that cause disease in people: P. falciparum, P. vivax, P. ovale, P. malariae. Plasmodium sp. may also infect primates, rodents, reptiles, and birds. Many birds can serve as a definitive host for these parasites. Plasmodium can be pathogenic to penguins, domestic poultry, ducks, canaries, falcons, and pigeons, but is most commonly carried asymptomatically by passerine birds. Avian malaria has a worldwide distribution and is of great economic significance is to the poultry industry. Organisms such as P. gallinaceum, P. juxtanucleare and P. durae may cause up to 90% mortality in poultry. Incidentally, birds with avian malaria have been used as model systems for studying the pathogenesis and treatment of malaria in humans. Clinical Signs Infected birds are often weak, depressed, dyspneic and anorexic. Birds may have abdominal protrusion (due to hepatosplenomegaly) and ocular hemorrhage. Central nervous system ischemia may occur with P. gallinaceum due to occlusion of capillaries by exoerythrocytic schizonts in heavily parasitized endothelial cells. Hemolytic anemia, often accompanied by leukocytosis, lymphocytosis, and hemoglobinuria, may also be present. Coma and death may occur quickly when the parasite burden is high. However, many birds, especially passerines, do not become ill and play an important role as asymptomatic carriers of the parasite. Life Cycle Plasmodium may exploit several genera of mosquitoes (Culex, Anopheles, Culiceta, Mansonia, and Aedes) as vectors and intermediate hosts. The mosquito inoculates the bird with sporozoites that enter the birds reticuloendothelial system. Each sporozoite develops into thousands of merozoites (preerythrocytic cycle) (Figure 1). These merozoites rupture their host cell and invade endothelial cells or other cells of the reticuloendothelial system to complete another cycle of replication. The merozoites then rupture that host cell and enter

erythrocytes in the blood stream. This initiates the intraerythrocytic cycle. Merozoites multiply in the RBC, forming a schizont (shizogony). The schizont will rupture, killing the RBC and releasing the merozoites to infect more RBCs. During schizogony, the parasites feed on the RBC cytoplasm, ingesting hemoglobin which produces brown pigment granules. The intraerythrocytic cycles continue until the host dies or the parasites are suppressed by host immunity. After the initial cycles in erythrocytes, a few merozoites develop into sexual cells (microgametes and macrogametes) with each new cycle. The sexual cells are maintained in the RBC until they are consumed by a mosquito with its blood meal.

Figure 1: Liver impression smear from a Penguin. Two macrophages containing numerous Plasmodium merozoites. The sex cells are released in the mosquitos midgut. Fertilization and zygote formation occurs when a microgamete encounters a macrogamete. The zygote matures into an elongated mobile cell that crosses the midgut wall. This cell is called an ookinete. The ookinete forms an oocyst on the outer wall of the midgut. The nucleus of the oocyst divides into thousands of spindle shaped sporozoites. The oocyst then bursts and releases the sporozoites, some of which migrate to the salivary gland where they are injected into a bird during the mosquitos blood meal. While most avian infections occur through the bite of a mosquito, it is possible for a direct bird-to-bird transmission to

occur. Schizogony occurs in the RBC and, therefore, blood-toblood transfer without the intermediate host can result in infection. The life cycle of avian malaria is very similar to that seen in infected human beings. However, birds (unlike mammals) suffer from repeated cycles of pre-erythrocytic merogony with reinvasion of reticuloendothelial cells by erythrocytic forms. Clinicopathologic Features and Diagnosis Experimental infection of domestic fowl resulted in peak parasitemia six days post-infection in most birds. The birds were febrile and anemic (hematocrit <24%). Approximately eight days post-infection, dysproteinemias characterized by hypoalbuminemia and a decreased -2 globulin concentration, with increased -1 and -2 globulins, were noted. Biochemical profiles revealed increased serum activities of aspartate aminotransferase, glutamate dehydrogenase, and glutamyltransferase, and a decreased creatinine concentration. Microscopic examination of a Wrights-stained thin blood smear is a sensitive method to detect Plasmodium intraerythrocytic trophozoites, schizonts or gametocytes. The trophozoite is a stage of the parasite that develops following invasion of the red blood cell by the merozoite. It is a small round to oval structure with a large vacuole that forces the parasite nucleus to one pole. This results in a signet-ring appearance (Figure 2). Schizonts are round to oval inclusions in the red cells containing darklystained merozoites.

Figure 2: Avian blood smear showing the intraerythrocytic, trophozoite stage of a malarial infection.

The intraerythrocytic gametocytes of Plasmodium can easily be confused with those of Haemoproteus because they both contain refractive yellow to brown pigmented granules (Figures 3 and 4).

Figure 3: Plasmodium gametocytes in avian blood. Note the smaller size when compared to Hemoproteus organisms.

Figure 4: Hemoproteus in avian blood. Typically larger than Plasmodium gametocytes and often have a characteristic canoe shape that wraps around the erythrocyte nucleus.

Features that can help distinguish between the two infections include: Plasmodium gametocytes are smaller in size than Haemoproteus organisms, usually occupying less than one-half of the host cell cytoplasm

Some Plasmodium gamonts (Haemoproteus does not)

displace

the

RBC

nucleus

Plasmodium may undergo schizogony in the peripheral blood (Haemoproteus does not) Plasmodium parasites can be found in cells other than erythrocytes (thrombocytes, leukocytes, and endothelial cells).

In birds that die peracutely, organisms may be few to sparse in the blood. In these cases, schizonts can be found in capillaries by examining impression smears of brain, lung, liver, and spleen. PCR has also been used to diagnose Plasmodium. However, this diagnostic test is most often used in a research setting. Treatment Note: Treatment of animals should only be performed by a licensed veterinarian. Veterinarians should consult the current literature and current pharmacological formularies before initiating any treatment protocol. Affected flocks can be treated with Chloroquine (5-10mg/kg) potentiated with primaquine (0.3mg/kg). Chloroquine can also be added to the drinking water at a dose of 250mg/120ml. Grape or orange juice may be needed to override the bitter taste of the medication. Quinacrine at a dose of 1.6 mg/kg given IM for 5 days is another treatment possibility. Additional treatments include sulfonamides combined with trimethoprim, pyrimethamine, and chlorguanil. Due to strain differences in susceptablity, different anti-malarial drugs can be tried. It is important to remember that malaria can be prevented by screening chicken houses to prevent contact with the mosquito vectors. References Aiello SE (ed): The Merck Veterinary Manual, 8th ed. National Publishing, Philadelphia, PA, 1998 Barriga O: Parasitology for Practitioners , 2nd ed. International Group, Minnesota, 1997. Campbell T: Avian Hematology and Cytology. Iowa State University Press, Ames, 1998. Kaufmann J: Parasitic Infections of Domestic Animals: A Diagnostic Manual, Birkhauser Verlag, 1996. Richie B, Harrison G, Harrison L: Avian Medicine: Principles and Applications. Wingers Publishing, Lake Worth FL, 1997. Slater LB. Malarial Birds: Modeling Infectious Human Disease in Animals. Bull. Hist. Med., 2005, 79: 261-294.

Williams RB: Avian Malaria: clinical and chemical pathology of Plasmodium gallinaceum in the domesticated fowl Gallus gallus. Avian Pathology February 2005 34(1), 29-47. Acknowledgement