Summary :Frank Hawking, in 1966 postulated that in synchronous malaria infections, the brief period of infectivity of gametocytes was timed to occur when the vector bites. Since this early work, numerous studies had contributed to confirm and explain this phenomenon with bird, rodent and primate Plasmodium. Data on the periodic production of gametocytes, the duration of their maturation, the effect of the schizogony on the infectivity and the circadian bioavailability of gametocytes provide some more informations on the periodic Plasmodium gametocyte infectivity to the vector. This paper is intended to be a review of contributions on the "Hawking phenomenon" and to summarize the principal causal hypotheses. The conclusion stresses the practical consequences for experimental studies and epidemiological surveys. A remarkable feature of most of bird, rodent and primate malaria is the precise timing of its recurrent attacks which are generally at some multiple of 24 hours. This implies that the duration of the erythrocytic asexual cycle is stable and that all the parasites behave synchronously, reaching schizogony at the same time. Cell division takes place at an hour of the day constant for each species of malaria parasite which depends on the location of the host or for some species mostly on the time of inoculation of the parasite. The biological purpose of such an accurate timing in the cycle of Plasmodium was shown by Hawking (1970) to assist them to present infective gametocytes at the time mosquitoes bite and

Direct feeding of Anopheles stephensi mosquitoes on mice infected with Plasmodium vinckei petteri showed that, during the periods of schizogony in the blood, the infectivity of gametocytes was markedly reduced. This could be prevented by prior injection of the L-arginine analogue, Nw-nitro-L-arginine (NwNLA) showing that the altered infectivity was due to reactive nitrogen intermediates (RNI). Similar effects on transmission of P. yoelii nigeriensis were demonstrated in vitro by membrane feeding of the mosquitoes. The in vitro reduction in infectivity could be reversed by injecting the L-arginine analogue either into the infected mouse donor of serum, or into the membrane feeding chamber. Elevated levels of TNF and IL-6 were demonstrated during the course of infection but did not correlate well with nitrogen radical activity. Similarly, direct measurements of NO2- and NO3- did not reflect the nitrogen radical activity revealed by addition of the specific L-arginine analogue.

The aim of this study was to determine the frequency of Toxoplasma infection in wild mammals, particularly noncarnivorous species, and to identify possible sources of infection for humans. A serological study was conducted that included 310 animals representing 10 species of terrestrial mammals from the same region (primary tropical rain forest inhabited by humans). Toxoplasmosis was diagnosed using a direct agglutination test using formalin-treated Toxoplasma gondii. The prevalence of antibodies for toxoplasmosis differed greatly between species and ranged from 0% to 62%. The seroprevalence of toxoplasmosis was zero or very low for tree-dwelling animals, which included sloth, porcupine, and howler monkey. Conversely, the prevalence of infection was 46-62% among granivores, insectivores, and burrowing animals, such as peccary, paca, and armadillo. We conclude that wild mammals in French Guiana constitute a reservoir of T. gondii parasites. Given the absence of domestic cats from the forest, noncarnivorous species may be infected by the ingestion of oocysts that are eliminated into the soil or water in the stools of wild cats and then dispersed.

We previously reported the identification of a T cell epitope in the N-terminal part of the circumsporozoite protein (CSP) of Plasmodium yoelii yoelii (Pyy). CD4+ T cell clones derived from mice immunized with a 21-mer peptide (amino acids 59-79, referred to as Py1) containing this epitope confer complete protection after passive transfer in mice. These clones proliferate in vitro in the presence of a 13-mer peptide (amino acids 59-71, referred to as Py1T). This shorter peptide was found to behave as a Th epitope in vivo, allowing overcoming of the genetic restriction for production of anti-repeat antibodies in BALB/c mice, when cross-linked to three (QGPGAP) repeats of the Pyy CSP. In this study, we report protection in BALB/c mice, against a challenge with Pyy sporozoites after immunization with linear and multiple antigen peptides containing Py1T as T epitope and three repeats QGPGAP (Py3) as B epitope. Multiple antigen peptide (MAP4-Py1T-Py3)-induced immunity was shown to be more effective than immunity induced by the linear form of the conjugate (Py1T-Py3), protecting against challenges with higher numbers of sporozoites. In both cases, levels of anti-repeat antibodies were strongly correlated with anti-parasite antibodies and protection. When tested in vitro, sera from mice immunized with the protective constructs strongly inhibited Pyy liver stages, while lymph node T cells displayed no cytotoxicity. In vivo, depletion of CD4+ or CD8+ T cells did not affect protection. Furthermore, MAP4-Py1T-Py3-immunized mice were not protected against a challenge with P. yoelii nigeriensis sporozoites, a parasite which has the same Py1T sequence but differs from Pyy in its repeated sequence. These results demonstrate that anti-repeat antibodies raised by immunization with the linear or the MAP form are exclusively responsible for the protection. Furthermore, this antibody response is boosted by a sporozoite challenge, allowing protection against a second challenge.