Exoerythrocytic forms of Plasmodium gallinaceum were cultured in vitro using salivary gland sporozoites extracted from experimentally infected Aedes fluviatilis mosquitoes. The host cells were macrophage precursors from chicken bone marrow. At various times after introduction of sporozoites, the cultures were stained by Giemsa or by immunofluorescence assay (IFA) using anti-sporozoite-specific monoclonal antibodies (MAb). The time to complete parasite development in vitro was 50-70 h. By 70 h, ruptured segmenters and free merozoites were visible within the cells. Inoculation of normal chickens with infected cultures induced parasitemia after a pre-patent period of 10-11 days. In vitro young exoerythrocytic forms, late schizonts that include the matured segmenters, and free merozoites shared common antigens with the sporozoites as revealed by IFA using anti-sporozoite-specific MAbs. Our data indicate that macrophages support development of P. gallinaceum sporozoites and that the circumsporozoite proteins are present until the end of the primary exoerythrocytic schizogony.

A correlation was observed between in vivo and in vitro activity of six monoclonal antibodies (mAb) against the major circumsporozoite protein of the avian malaria Plasmodium gallinaceum as follows. (1) Two mAb were protective, totally abrogating sporozoite infectivity to chicks, its natural host, in vivo; they caused 100% inhibition of sporozoite invasion (ISI) in vitro to SL-29 chicken fibroblasts and intense ISI to cultured chicken macrophages, as well as inhibited the exoerythrocytic development of sporozoites taken up by macrophages, the initial cell host of P. gallinaceum sporozoites. (2) Two mAb were partially protective in that they reduced sporozoite infectivity to chicks, caused partial ISI to SL-29 and macrophage cells and partial inhibition to the exoerythrocytic development of sporozoites in macrophages in vitro. (3) Two mAb were totally inactive in vivo although they both bound to the sporozoite antigens as detected by indirect immunofluorescence, western blot, and ELISA; they both failed to induce ISI or inhibit the exoerythrocytic development in macrophages. The possible participation of macrophages as the initial cell type involved in sporozoite destruction in the presence of anti-circumsporozoite antibodies is discussed.

The aim of this research was to compare two artificial insemination protocols (AIP): hormonal synchronization with fixed time artificial insemination (SC-FTAI) and the use of a table based on visual observation of estrus signs (VO) in order to identify cows in natural or spontaneous estrus being assigned to AI (NSE-IA). Two groups were formed: in the first group 109 cows were assigned to SC-FTAI, in which a commercial protocol is used; the second one included 108 randomly chosen cows, which were assigned to NSE-AI and in this group a modified table was used. Response variable was first service fertility rate (FSF), which was coded 1 for pregnant and 0 for empty. Predictor variables were AIP, postpartum anestrus, daily milk yield, body condition score at AI and calving number. Statistical analyses included association chi-square tests and logistic regression. Results showed an overall 41.94% FSF and a significant association was detected (P < 0.05) between FSF and daily milk yield; pregnancy rates were 42.20% and 41.67% for the SC-FTAI and NSE-IA groups, respectively (P > 0.05). The odds ratio for the effect of AIP was only 1.050, suggesting no differences in FSF between groups. The NSE-AI protocol can enhance both the technique of VO and reproductive efficiency. Further validation of the table is required.