Professor L. J. Bruce-Chwatt's 80th birthday.
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Biomedical subjects
Publications and source records attributed to P C Garnham.
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In a continuing reexamination of plasmodial tissue stages within the context of the hypnozoite theory of malarial relapse, 2 strains of Plasmodium vivax with distinct and disparate relapse characteristics in humans were studied in chimpanzees. Following intravenous inoculation of massive numbers of salivary gland sporozoites, both the frequently relapsing Chesson strain and a North Korean strain characterized by predominantly delayed relapses exhibited relapse patterns and antimalarial sensitivity in the splenectomized chimpanzee essentially indistinguishable from those seen in humans. Examination of hepatic biopsies obtained at 7 and 10 days after infection revealed both pre-erythrocytic (pre-e) schizonts and hypnozoites in tissue obtained from the animal infected with the Chesson strain, but only rare hypnozoites (no pre-e schizonts) at 7 days in the animal infected with the North Korean strain. These findings, combined with the comparability of relapse behavior--which indicates the suitability of the chimpanzee as a model for the natural (human) host-parasite relationship--are essentially as predicted by the hypnozoite theory, despite the small numbers of tissue forms seen. Pre-erythrocytic schizogony of the Chesson strain in the liver was essentially indistinguishable from that of other strains studied, also underlining the suitability of this model system for tissue stage studies of P. vivax.
Rhesus monkeys were heavily infected with sporozoites of Plasmodium cynomolgi bastianellii in an attempt to demonstrate the site of invasion of sporozoites into tissue cells and their growth there. Further attempts were made to correlate the appearance and loss of hypnozoites with parasitaemic relapses. Hypnozoites were demonstrated and once again shown to decrease in numbers over 229 days during which time the infection showed parasitaemic relapses. Liver biopsies taken at two-day intervals for 12 days showed that hypnozoites decreased in numbers over-all and growing schizonts were demonstrated in the liver. At this time a parasite the size of a hypnozoite was seen with two nuclei and another was seen with an elongate, possibly dividing nucleus in one monkey. an attempt to find the location of the early intracellular exoerythrocytic forms in the liver at various times less than 40 hours after infection using smears and immunological staining with newly prepared anti-sera failed. Large numbers of sporozoites of P. knowlesi were also injected into a rhesus monkey the liver of which on the fifth day after infection showed no hypnozoites among 157 sections of growing schizonts and no parasites at all on the 42nd day after infection. In P. cynomolgi bastianellii infections parasites, mostly hypnozoites, were found in the liver up to 229 days after infection.
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A new species of malaria parasite, Plasmodium (Sauramoeba) heischi, is described from African skinks (Mabuya striata). Eleven individuals of 90 specimens collected in Nairobi were found to be infected. The new parasite is a large species, characterized by spindle-shaped gametocytes, the female often with a subterminal nucleus. The schizonts produce up to 65 nuclei and cause great hypertrophy and distortion of the host cell. Although similar to P. (Sauramoeba) giganteum in dimensions and merozoite numbers, P. heischi is easily distinguished by gametocyte and schizont shapes.
Hypnozoites of two strains of the human relapsing malaria parasite, Plasmodium vivax, have been detected among maturing 7- and 10-day pre-erythrocytic schizonts in liver biopsies of chimpanzees infected by intravenous inoculation of sporozoites obtained from dissected salivary glands of heavily infected anopheline mosquitoes. As in the simian relapsing species, P. cynomolgi, the hypnozoites of P. vivax at 7 and 10 days are uninucleate forms of approximately 4-5 micrometers diameter, lying within the cytoplasm of individual hepatocytes. Their presence in this relapsing human species is added support for the hypnozoite theory of malarial relapse.
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With the exception of the human malaria parasites, the mammalian species of Plasmodium have a fairly restricted geographical distribution, while the incidence of the infection in the different orders of mammals is patchy. A few species have been described in ungulates in the Old World Tropics, but Plasmodium odocoilei sp.nov. of the white-tailed deer (Odocoileus virginianus) of Texas is the first example to be found in the New World. It was discovered in an adult specimen that had had its spleen removed. Only the blood stages of the parasite are known, but these show various remarkable features: the presence of large vacuoles in the cytoplasm, and the enormous distortion and discoloration of the infected red blood cell. The phylogeny of the parasite is discussed in relation to that of the vertebrate host and it is suggested that Cervid stock crossed the Bering Land Bridge in the Pliocene Age, carrying a plasmodial infection that was the common source of malaria in the Old World Tragulidae and in the New World Odocoileus.
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The North Korean strain of Plasmodium vivax was characterized by its extraordinarily prolonged incubation period in certain circumstances. It was clearly demonstrated by quantitative observations that the phenomenon may be elicited by the inoculation of small numbers of sporozoites. After the intradermal inoculation of 10 or 100 sporozoites, the incubation period was delayed for periods varying between 262 and 628 days; after u,000 sporozoites, with one exception, the same delay occurred; after 100,000 sporozoites, the incubation period was always of normal duration (13--16 days). Two laboratory-acquired infections in workers who had taken a prophylactic drug showed incubation periods of 315 and 329 days, respectively. Various theories for the phenomenon of prolonged prepatent periods are examined, and the most satisfactory one is based on the presumed existence of two populations of sporozoites in P. vivax. In temperate strains, sporozoites requiring long prepatent periods (LPP) for development are present in great excess over a much smaller proportion of sporozoites characterized by short prepatent periods (SPP); thus small doses will elicit the phenomenon, though doses of over 1,000 sporozoites will mask the effect as the few SPP sporozoites will produce an infection with a normal (i.e. short) prepatent period. In tropical strains, the relative proportions are different, perhaps in equal numbers, and even in small doses some SPP sporozoites will be present and normal prepatent periods should ensure whatever the dosage."