Freeze-Preservation of Leucocytozoon simondi Sporozoites in Liquid Nitrogen

SYNOPSIS. Sporozoites of L. simondi were maintained in a viable state for 7 months in liquid nitrogen. Comparison of parasite development initiated with fresh and frozen sporozoites showed a delay in development of each stage studied. Comparisons of prepatency, first elongate gametocytes, peak density of round and elongate forms, anemia and disappearance of megaloschizonts were made. In each phase there was a delay of 2–3 days in ducks infected with frozen sporozoites.     

The fine structure of elongate gametocytes of Leucocytozoon ziemanni (Laveran).

In an effort to establish comparative data within the genus Leucocytozoon, elongate gametocytes of L. ziemanni from naturally infected great horned owls (Bubo virginianus) were examined by electron microscopy. Micro- and macrogametocytes proved to be easily distinguishable at the electron microscopic level due to dramatic dimorphism at maturity and cytoplasmic and nuclear morphology. The parasite membrane architecture, number and type of cytoplasmic ribosomes of both micro- and macrogametocytes, presence and arrangement of osmiophilic bodies and electron dense spheres, mitochondrial morphology, endoplasmic reticulum cisternae morphology, mitochondria containing pocket infoldings of the nuclear membrane of the microgametocytes, and cytostome and food vacuole formation compare favorably with available information on L. simondi and L. smithi. Comparative variations exist only in that L. ziemanni gametocytes apparently lack compartmentalization of the cytoplasm by aligned unit membranes and parasite induced separations of the host cell nucleus as reported for L. simondi.     

Anemia in Ducks Infected with Leucocytozoon simondi*†

SYNOPSIS. Thirty-four white Pekin ducklings were used to study the anemia associated with infection by Leucocytozoon simondi. The first appearance of gametocytes in the peripheral blood, as detected by thin smears, marked the onset of anemia. This anemia lasted for the duration of the initial parasitemia, usually reaching a low point early in the infection (1 to 5 days post patency) and returning slowly to normal as the parasitemia decreased. Greatest gametocyte density occurred 5 to 8 days post patency. In a number of cases recovery from anemia began simultaneously or even prior to the highest level in the gametocyte density. In low level parasitemias a fluctuation occurred in erythrocyte numbers which corresponded with the peaks of gametocyte density. In none of the infections was a sufficient number of parasite observed to account for the existing anemia. Haemopoietic activity was observed for a brief period at the time of maximum erythrocyte loss in only a few birds. The overcompensation for erythrocyte loss at the end of the primary parasitermia favors the view that increased erythrocyte production may account for the short duration of haemopoiesis.     

Experimental Studies of the Life Cycle of Leucocytozoon simondi in Ducks in Norway*

SYNOPSIS. Pekin ducks were infected naturally and experimentally. The life cycle of Leucocytozoon simondi was followed in the ducks and the vector, a new species of Simulium, close to Simulium dogieli. The prepatent period in the ducks was 4–5 days and developing megaloschizonts appeared in 6–7 days. Schizonts were found in liver, spleen, brain, and kidney. In the kidneys they were located in the glomeruli. Sporogony in some individuals was completed in 7 days at 13–14 C, other individuals developed more slowly, as ookinetes were found in flies 8 days after feeding. The rapid asexual cycle combined with a sporogonic cycle, in which some ookinetes develop rapidly and others more slowly, favors the maintenance of the parasite in an environment with relatively low daily temperatures. This and the size of the megaloschizonts indicate differences between this strain of the parasite and the one occurring in North America.     

Schizogony and Gametogony of Leucocytozoon simondi and Associated Reactions in the Avian Host*

SYNOPSIS. Stages of development of Leucocytozoon simondi in White Pekin ducklings and their reactions to the parasite were studied on successive days after infecting them artificially with sporozoites from Simulium rugglesi. The minimum prepatent period was 5 days. The first asexual cycle occurred exclusively in the parenchymal cells of the liver. Progeny of these hepatic schizonts followed one of 3 courses: (a) invaded parenchymal liver cells to give rise to another hepatic cycle, (b) penetrated blood cells to form round gametocytes, and (c) were phagocytized by macrophages and grew into megaloschizonts thruout the body. The appearance of elongating gametocytes coincided with the period of maturation and release of merozoites from the megaloschizonts. Experimental evidence supports the hypothesis that the round gametocytes arise from the hepatic schizonts and the elongate forms from the megaloschizonts. Mature megaloschizonts released millions of merozoites, but a high 2nd peak in parasitemia did not develop because of retention of developing gametocytes in the deep circulation, particularly the liver and spleen, and a pronounced host reaction.     

The fine structure of the developing oocyst of Pterospora floridiensis (Apicomplexa,Urosporidae)

Developing oocysts of the gregarine Pterospora floridiensis Landers 2001 were examined by transmission electron microscopy. Each oocyst had an outer capsule and an inner capsule that contained 8 sporozoites. In early stages of development the inner capsular wall was separated from the developing sporozoites and residual mass, and was not appressed to the sporozoites. Early stage sporozoites were connected to a residual mass and were filled with endoplasmic reticulum, golgi and numerous developing secretory vesicles. In late stages of oocyst and sporozoite development, the inner capsular wall was closely appressed to the sporozoite surface. The inner capsular wall was ～60-100 nm thick and the outer capsular wall was ～160-320 nm thick. There were no extensions on the outer wall for which the genus was named. Late stage sporozoites had no residual mass connection, were more electron dense, and contained three distinct types of dense secretory structures: 1) small oval/spherical dense vesicles, 2) large (350-400 nm) vesicles near the anterior end, and 3) elongated dense tubular bodies that converged at the apex. Few ultrastructural reports exist of developing gregarine oocysts and sporozoites, and as more studies are completed these morphological characteristics may be important in interpreting molecular phylogenetic analyses.