Infectivity of Plasmodium gallinaceum Sporozoites from Oocysts

SYNOPSIS. Infectivity of Plasmodium gallinaceum (Brumpt) sporozoites isolated from midguts and salivary glands of experimentally infected Aedes fluviatilis (Lutz) was studied. The 2 populations were compared at 7, 8, and 9 days postisolation from mosquitoes, which were maintained at 27 C ± 1C and ∼75% relative humidity. Infectivity of the parasites was evaluated by the length of the prepatent period of the infection in 2-week-old chicks inoculated intramuscularly. Infection was caused by 7-day-old sporozoites from salivary glands, but not from midguts. Older sporozoites induced infection in all the inoculated chicks. The results suggested a somewhat higher infectivity of the 8- and 9-day salivary-gland parasites than of the oocyst sporozoites. However, unlike sporozoites from mammalian malaria, oocyst sporozoites from avian malaria were highly infective at this age.     

Plasmodium gallinaceum: mechanisms of anemia in infected chickens

Erythrocyte labeling by random and cohort techniques was used to study erythrocyte survival in normal chickens and chickens infected with Plasmodium gallinaceum. Occurrence of erythrocyte destruction during the prepatent period was apparent in infected chickens by both techniques. Treatment with the antimalarials chloroquine and quinacrine not only cleared the circulation of parasites promptly but brought about an equally prompt cessation of disease-related erythrocyte destruction. Plasmodium gallinaceum infection caused a transitory decrease in blood volume at the time of rapid decrease in packed cell volume. The blood volume returned to preinfection values before the packed cell volume returned to normal. Parasitized erythrocytes were present in capillaries of the spleen, liver, and bone marrow during the entire prepatent period of the infection, thus providing a reasonable explanation for erythrocyte destruction observed in the absence of parasitemia during the prepatent period.     

Plasmodium gallinaceum: sporozoite activity in immune mosquito hemolymph

Sporozoites of Plasmodium gallinaceum are relatively inactive in Hanks' balanced salt solution or in the hemolymph of the susceptible Aedes aegypti mosquito. They become agitated and very active in the presence of the hemolymph of the innately immune mosquito, Culex pipiens. Sporozoites from the latter are more infective in chicks than the former. This behavior is likely the result of stimulation or irritation by the adverse environment.     

Exoerythrocytic development of Plasmodium gallinaceum in the White Leghorn chicken

Plasmodium gallinaceum typically causes sub-clinical disease with low mortality in its primary host, the Indian jungle fowl Gallus sonnerati. Domestic chickens of European origin, however, are highly susceptible to this avian malaria parasite. Here we describe the development of P. gallinaceum in young White Leghorn chicks with emphasis on the primary exoerythrocytic phase of the infection. Using various regimens for infection, we found that P. gallinaceum induced a transient primary exoerythrocytic infection followed by a fulminant lethal erythrocytic phase. Prerequisite for the appearance of secondary exoerythrocytic stages was the development of a certain level of parasitaemia. Once established, secondary exoerythrocytic stages could be propagated from bird to bird for several generations without causing fatalities. Infected brains contained large secondary exoerythrocytic stages in capillary endothelia, while in the liver primary and secondary erythrocytic stages developed primarily in Kupffer cells and remained smaller. At later stages, livers exhibited focal hepatocyte necrosis, Kupffer cell hyperplasia, stellate cell proliferation, inflammatory cell infiltration and granuloma formation. Because P. gallinaceum selectively infected Kupffer cells in the liver and caused a histopathology strikingly similar to mammalian species, this avian Plasmodium species represents an evolutionarily closely related model for studies on the hepatic phase of mammalian malaria.     

Motility and infectivity of Plasmodium berghei sporozoites expressing avian Plasmodium gallinaceum circumsporozoite protein

Avian and rodent malaria sporozoites selectively invade different vertebrate cell types, namely macrophages and hepatocytes, and develop in distantly related vector species. To investigate the role of the circumsporozoite (CS) protein in determining parasite survival in different vector species and vertebrate host cell types, we replaced the endogenous CS protein gene of the rodent malaria parasite Plasmodium berghei with that of the avian parasite P. gallinaceum and control rodent parasite P. yoelii. In anopheline mosquitoes, P. berghei parasites carrying P. gallinaceum and rodent parasite P. yoelii CS protein gene developed into oocysts and sporozoites. Plasmodium gallinaceum CS expressing transgenic sporozoites, although motile, failed to invade mosquito salivary glands and to infect mice, which suggests that motility alone is not sufficient for invasion. Notably, a percentage of infected Anopheles stephensi mosquitoes showed melanotic encapsulation of late stage oocysts. This was not observed in control infections or in A. gambiae infections. These findings shed new light on the role of the CS protein in the interaction of the parasite with both the mosquito vector and the rodent host.     

Fine structure of Plasmodium gallinaceum in embryonic and neonate chicks.

The erythrocytic stages of Plasmdoium gallinaceum in chicken embryos injected with parasited blood either from a syringe-passaged infection in chickens or from a chicken infected with sporozoites were characterized by abnormal structure. Particularly evident were large, unstained vacuoles within the cytoplasm; these occurred with greatest frequency in schizonts. The presence of myelin bodies within these vacuoles was revealed by transmission electron microscopy; abnormal cytokinesis and aberrant merozoites provided additional evidence of the parasite's inability to develop naturally within the milieu of the embryonic erythrocytes. Fifty-five passages were necessary to restore normal structure of the parasites in embryos, while only 5 passages were required for such restoration in neonate chicks. The probable adaptation of the parasite to the proportions of hemoglobin of the adult chicken may be responsible for the abnormal growth in the immature host.