Aspectos parasitários observados no local inoculado com esporozoitos de Plasmodium Gallinaceum

The following is a summary of the studies made on the development of Plasmodium gallinaceum sporozoites inoculated into normal chicks. Initially large numbers of laboratory reared Aëdes aegypti were fed on pullets heavily infected with gametocytes. Following the infectious meal the mosquitoes were kept on a diet of sugar and water syrup until the appearance of the sporozoites in the salivary glands. Normal chicks kept in hematophagous arthropod proof cages were then inoculated either by bite of the infected mosquitoes or by subcutaneous inoculations of salivary gland suspensions. By the first method ten mosquitoes fed to engorgement on each normal chick and were then sacrificed immediately afterwards to determine the sporozoite count. By the second method five pairs of salivary glands were dissected out at room temperature, triturated in physiological saline and inoculated subcutaneously. The epidermis and dermis at the site of inoculation were excised from six hours after inoculation to forty eight hours after appearance of the parasites in the blood stream and stretched out on filter paper with the epithelial surface downward. The dermis was then curretted. Slides were made of the scrapings consisting of connective tissue and epithelial cells of the basal layers which were fixed by metyl alcohol and stained with Giemsa for examination under the oil immersion lens. Skin fragments removed from normal chicks and from regions other than the site of inoculation in the infected chicks were used as controls. In these, only the normal histological aspect was ever encountered. In the biopsy made at the earliest period following inoculation clearly defined elongated forms with eight or more chromatin granules arranged in rosary formation were found. The author believes these to be products of the sporozoite evolution. Search for transition stages between these forms and sporozoites is planned in biopsies to be taken immediately following inoculation and at given intervals up to the six hour period. 1.) 6 and 12 hour periods. The bodies referred to above found in the first period in great abundance, apparently in proportion to the large numbers of sporozoites inoculated, were perceptibly reduced in numbers in the second period. 2.) 18 hour period. Only one biopsy was examined. This presented a binuclear body shown in Fig. 1, having a more or less hyaline protoplasm staining an intense blue and a narrow vacuole delimiting the cell boundaries. The two chromatin grains were quite large presenting a clearly defined nuclear texture. 3.) 24 hour period. A similar body to that above (Fig. 2) was seen in the only preparation examined. 4.) 60 hour period. The exoerythrocytic schizonts were found more frequently from this period onward. Several such were found no longer to contain the previously described vacuoles (Fig. 3). 5.) 84 hour period. Cells bearing eight or more schizonts were frequently encountered here. That these are apparently not bodies in process of division may be seen in Fig. 4. From this time onward small violet granules similar to volutine grains appeared constantly in the schizont nucleus and protoplasm. These are definitely not hemozoin. The above observations fell within the incubation period as repeated examinations of the peripheral and visceral blood were negative. Exoery-throcytic parasites also were never encountered in the viscera at this time. Exoerythrocytic schizonts searched for at site of inoculation 1, 24 and 48 hours after the incubation period were present in large number at all three times with apparent tendency to diminish as the number within the blood stream increased. Many of them presented the violet granules mentioned above. The appearance of the chromatin and the intensity of staining of the protoplasm varied from body to body which doubtless corresponds to the evolutionary stage of each. This diversity of aspect may frequently be seen in the parasites of the same host cell (Fig. 5.). These findings lend substance to the theory that the exoerythrocytic forms are the link between the sporozoites and the pigmented parasites of the red blood corpuscles. The explanation of their continued presence in the organism after infection of the blood stream takes place and their presence in cases infected by the inoculation blood does not come within the scope of this work. Large scale observations shortly to be undertaken will be reported in more detail particularly observations on the first evolutionary phases of the sporozoite within the organism of the vertebrate host.

Infecção experimental do Culex quinquefasciatus pelo Plasmodium juxtanucleare

Foram feitas duas séries de experiências para verificar a possibilidade da infecção de mosquitos com o P. juxtanucleare. Na primeira foram utilizados 158 exemplares de Aedes aegypti e 152 de Culex quinquefasciatus, com resultados negativos. Na segunda série foram usados 15 espécimes de Aedes lepidus, com resultado negativo, e 80 de C. quinquefasciatus. Dêstes últimos apenas 3 se infectaram, não tendo sido possível transmitir a infecção a pintos inoculados. Êste culicíneo não é considerado transmissor habitual do plasmódio em estudo em vista do baixo índice de infectividade observado nas experiências. São brevemente discutidos os resultados do presente trabalho, inclusive o insucesso das inoculações em pintos.

A transmissão de Plasmodium Gallinaceum pelo Aedes (Ochlerotatus) lepidus

Foi pesquisada a capacidade transmissora do Aëdes (O.) lepidus em relação ao Plasmodium gallinaceum. Êste mosquito comportou-se como ótimo vector, apresentando elevado índice oocístico e esporozoítico e produzindo a malária por picada em animais sensíveis (pintos e frangos).

Estudos sôbre índice de siclemia

O índice de sinclêmicos encontrado foi de 10%, examinando-se uma amostra heterógenea de 1130 indivíduos de grupo étnico negróide e mestiços desta etnia. Foi negativa a pesquisa em 120 indivíduos caucasóides e 30 mongolóides. O "índice familiar de siclêmicos" - a percentagem desses indivíduos em um grupo de famílias - fornece um resultado mais proximo do verdadeiro número de siclêmicos em uma amostra de população. O método clássico fornece um número inferior ao existente na amostra estudada. A aplicação do teste de siclemia em Jurisprudência, Antropologia e Etnografia, poderá contribuir, na qualidade de método auxiliar, no estudo de vários problemas. São indicados como métodos de escolha: o de Emmel e o de maceração de tecidos conservados em solução de formol ou alcool. Êste último permite estudos retrospectivos. A estase circulatória parece não ser fator suficiente para a passagem da condição de siclêmico para anemico. O parasitismo da hemátia pelo Plasmodium vivax não impede o fenômeno da siclisação. Infecção por plasmodídeos (P. falciparum e P. vivax) não é fator suficiente para desencadear a doença. A importância da descoberta de Emmel (descoberta do fenômeno da siclisação) reside no conhecimento em doença heredofamiliar de fato novo de máxima significação: a possibilidade de identificar os indivíduos ainda na fase de latência, ou seja, no período que medeia entre o nascimento e a instalação dos primeiros sintomas da doença. A relação entre casos de anemia falciforme ocorridos em gerações por vêzes muito afastadas (fenômeno denominado "skipping" - salto - pelos autores de lingua inglêsa) é mantida através dos siclêmicos. Êsses indivíduos podem ser comparados aos portadores de germe no sentido empregado na profilaxia das doenças infectuosas. Exames periódicos dos siclêmicos poderiam contribuir para esclarecimento da patogenia e forneceriam dados úteis para a profilaxia da doença. Seriam de grande alcance medidas idênticas em outras doenças hereditárias...

Infecção do macaco sul americano "saguí" (Callitrix jacchus, Linneu 1758) com o Plasmodium Knowlesi

Foram observadas infecções no macaco americano "Sagui" (Callitrix jacchus, Linneu 1758), quando inoculados com Plasmodium knowlesi. Inoculações massiças intra-venosas, são mortais em cerca de 10 dias. Com inoculações intra-musculares de menores quantidades de parasitos, foram verificados alguns casos de curas espontaneas. As características morfológicas e o seu conhecido ciclo de 24 horas taes como observados na infecção do macaco Rhesus, permanecem as mesmas no macaco estudado. O vigor do "sagui" em cativeiro é nitidamente inferior ao do "Rhesus" mas como seu preço de custo é trinta vezes menor, deve ser êle considerado como mais um animal útil para as pesquisas em malaria experimental.

Factors influencing the development of Plasmodium gallinaceum in Aedes fluviatilis

Aedes fluviatilis is susceptible to infection by Plasmodium gallinaceum and is a convenient insect host for the malaria parasite in countries where Aedees aegypti cannot be maintained in laboratories. In South America, for instance, the rearing of A. aegypti the main vector of urban yellow fever, is not advaisable because of the potential health hazard it represents. Our results of the comparative studies carried out between the sporogonic cycle produced with two lines of P. gallinaceum parasites into A. fuviatilis were as follows. As proved for A. aegypti, mosquito infection rates were variable when A. fluviatilis blood-fed on chicks infected with and old syringe-passaged strain of P. gallinaceum. Oocysts developed in 41% of those mosquitos and the mean peak of oocyst production was 56 per stomach. Salivary gland infections developed in about 6% of the mosquitos. The course of sporogony was unrelated to the size of the inoculum administered to chicks or to the route by which the birds were infected. The development of infected salivary glands was unrelated to oocyst production. Sporogony of P. gallinaceum was more uniform when mosquitos blood-fed on chicks infected with a sporozoite-passaged strain. Oocysts developed in about 50% of those mosquitoes and the mean peak of oocyst production was 138 per stomach, with some individuals having as many as 600-800 oocysts. Infected salivary glands developed in a mean of 27% of the mosquitos but, in some batches, was a high as 50%. Patterns of salivary gland parasitism were similar to those of oocyst production. The course of sporogony of P. gallinaceum in A. fluviatilis is analized in relation to degree of parasitemia and gametocytemia in the vertebrate host.

Presença do Plasmodium brasilianum em macacos capturados na área de enchimento do reservatório da usina hidrelétrica de Tucuruí, Pará

Examinando-se o sangue de 2.046 primatas capturados durante a "Operação Curupira" encontraram-se plasmódios semelhantes ao Plasmodium brasilianum em sete espécies ou subespécies: Alouatta belzebul belzebul, Alouatta belzebul nigerrima, Alouatta seniculus, Chiropotes satanas, Callicebus moloch, Saimiri sciureus e Saguinus midas niger. Esta última espécie havia sido ainda encontrada naturalmente parasitada por plasmódios.