Prenatal toxoplasmosis diagnosis from amniotic fluid by PCR

Toxoplasmosis is one of the most common infections all over the world. Most cases are asymptomatic, except in immunosuppressed individuals and fetuses, which can be seriously damaged. Prenatal diagnosis should be made as soon as possible since treatment of the mother can minimize fetal sequelae. Our aim in this study was to test the polymerase chain reaction technique (PCR) in 86 samples of amniotic fluid from women who seroconverted during pregnancy. DNA was amplified using external primers and, in a second step, internal primers, in a nested PCR system. Samples were also inoculated into mice and the newborn were evaluated by T. gondii serology, skull x-ray, transfontanel ultrasound, fundoscopic examination, lumbar puncture and clinical examination. PCR was positive in seven cases and negative in 79. Among PCR-positive cases, two were negative by inoculation into mice and by clinical evaluation; among PCR-negative ones, three had clinical evidence of toxoplasmosis and one was positive after inoculation into mice. PCR showed values of sensitivity = 62.5% and specificity = 97.4%; the values of inoculation into mice where 42.9% and 100%, respectively. Although PCR should not be used alone for prenatal diagnosis of congenital toxoplasmosis, it is a promising method and deserves more studies to improve its efficacy.

Estudo prospectivo de gestantes e seus bebês com risco de transmissão de toxoplasmose congênita em município do Rio Grande do Sul

A população estudada foi composta por 2.126 gestantes atendidas em unidades do Sistema Único de Saúde da região noroeste do Estado do Rio Grande do Sul. Após o screening sorológico inicial ocorreu o acompanhamento das gestantes, durante o pré-natal, e de seus bebês. Foram realizadas dosagens de IgG, IgM, IgA, Avidez de IgG, inoculação em camundongos, PCR e coleta de placenta e de cordões umbilicais para realizar a técnica de imuno-histoquímica além de avaliações clínicas. Das gestantes avaliadas, 74,5% eram IgG reagentes e 3,6% IgM reagentes. Nas avaliações oftalmológicas, foi observada lesão em dez gestantes e uma criança apresentou lesões oftalmológicas e calcificações cerebrais. A presença de IgM específico anti-T.gondii, durante toda a gestação não caracterizou a fase aguda recente da infecção, fazendo-se necessária a realização de testes complementares. Ressalta-se a importância do acompanhamento de neonatos de mães com sorologia compatível com a infecção mesmo sem sinais e sintomas sugestivos de toxoplasmose congênita.

Toxocariasis of the central nervous system: with report of two cases

Clinical involvement of the nervous system in visceral larva migrans due to Toxocara is rare, although in experimental animals the larvae frequently migrate to the brain. A review of the literature from the early 50's to date found 29 cases of brain involvement in toxocariasis. In 20 cases, various clinical and laboratory manifestations of eosinophilic meningitis, encephalitis, myelitis or radiculopathy were reported. We report two children with neurological manifestations, in which there was cerebrospinal fluid pleocytosis with marked eosinophilia and a positive serology for Toxocara both in serum and CSF. Serology for Schistosoma mansoni, Cysticercus cellulosae, Toxoplasma and cytomegalovirus were negative in CSF, that was sterile in both cases. Improvement of signs and symptoms after specific treatment (albendazole or thiabendazole) was observed in the two cases. A summary of data described in the 25 cases previously reported is presented and we conclude that in cases of encephalitis and myelitis with cerebrospinal fluid pleocytosis and eosinophilia, parasitic infection of the central nervous system should be suspected and serology should be performed to establish the correct diagnosis and treatment.

Toxoplasmose humana: meningoencefalomielite toxoplasmica: ocorrência em adulto e recemnascido

Desde 1914, com o trabalho inicial de Castellani, pesquisadores de diversos países teem assinado o encontro de toxoplasmose humana, o que foi até recentemente objeto de controvérsias. Como resultado dos trabalhos de Torres, Levaditi & coL, Wolf, Cowen & Paige, Pinkerton & Henderson e Sabin, a questão do parasitismo do homem por Toxoplasma, deve ser encarada considerando-se que teem sido descritos casos duvidosos e casos não duvidosos: a) Duvidosos, são os casos referidos por Castellani (Ceilão, 1914); Fedorovitch (Mar Negro, 1916); Chalmers & Kamar (Sudão, 1920); e Bland (Londres, 1930-31). b) Não duvidosos são os relatados por Janku (Praga, 1923); Torres, (Rio de Janeiro, 1927); Wolf & Cowen (New-York, 1937); Richter (Chicago, 1936: diagnósticado por Wolf & Cowen em 1938); Wolf, Cowen & Paige (New-York, 1939); Hertig (Massachusetts, 1935: diagnósticado por Pinkerton & Weinman em 1940); Pinkerton & Weinman (Lima-Peru, 1940); Sabin (dois casos, Cincinnati, 1941); Pinkerton & Henderson (dois casos, St. Louis, 1941); Paige, Cowen & Wolf (três casos, New-York, 1942); e De Lange (Amsterdam, 1929: diagnósticado por Paige, Cowen & Wolf em 1942). Destes relatos considerados como casos não duvidosos de toxoplasmose humana, 10 representam uma doença congênita (Torres e Paige, Wolf & Cowen) ocorrendo em crianças recemnascidas ou de poucos meses de vida. Como nas vezes em que foi possivel examinar as mães dessas crianças tratava-se de mulheres com aspecto sadio, concluiu-se que a moléstia, embora sumamente grave e mortal, pode apresentar uma forma inaparente. Em dois casos, provas de soro proteção demonstraram essa infecção materna inaparente, pelo encontro de anticorpos neutralizantes para Toxoplasma (Paige, Cowen & Wolf). Nada obstante não ter sido feito um estudo microscópico detalhado em todos os casos, a doença caracteriza-se por uma meningoence falomielite granulomatosa (Wolf & Cowen), associada frequentemente à miocardite e corioretinite. Os sintomas apresentados pelos pacientes, foram: febre, convulsões, distúrbios respiratórios, hidrocefalia, cianose, vômitos, labidade de temperatura, etc., sendo sinal diagnóstico de máxima importância, o encontro de focos de calcificação cerebral profunda e corioretinite em recem nascido (Dyke, Wolf, Cowen, Paige & Caffey) . Em dois casos de toxoplasmose verificados em pacientes de seis e oito anos respectivamente, foi encontrada uma encefalite clinicamente atípica, sendo que um deles sarou (Sabin). Os três casos restantes foram descritos em adultos; num deles as lesões não eram típicas (havia concomitância de infecção por Bartonella bacilliformis) e nos outros dois, a doença assumiu uma "forma exantemática", simulando as febres maculosas (Pinkerton & Henderson), sendo o achado mais importante microscopicamente, uma pneumonia intersticial. Alem da semelhança clínica, tambem os focos de encefalite encontrados nessa "forma exantemática" da toxoplasmose, são semelhantes aos descritos nas...

Miocardite na forma aguda da doença de Chagas

The autopsy of a case of CHAGAS'S disease or American tryponosomiasis (a girl, 5 years old), dead in the 22nd day of illness is reported. The anatomic diagnosis was a follows: Acute diffuse chagasic nyocarditis. Chagasic encephalitis. Chagasic lymphadenitis of the right posterior auricular node. Tuberculosis of the bronchial and pulmonary nodes. Chronic passive hyperemia and atelectasia of the lungs. Chronic passive congestion and hemorrhages of the spleen. Serous hepatitis. Parotiditis. Edema of the right eyelids. Bilateral hydrothorax. Hydropericardium. Hydroperitoneum. The morphology of Schizotrypanum cruzi in the myocardium is considered. Besides agglomerates with typical small oval or round intracellular bodies, pre-flagellate and flagellate organisms, others are found in which the great amount of parasites and marked pressure exerted by them against each other render very difficult their identification; sometimes the similitude of such agglamerates to Toxoplasma is striking (Fig. 1 and 1 A). In such a case, the structure of the blepharoplast (Fig. 1 and IA), usually preserved, is profitable and allows the identification of the pre-flagellate and flagellate forms of Schizotrypanum cruzi. Most of the small sensitive nerves in the epicardium shows mononuclear infiltration of the perineurium (perineuritis, Figs. 12-14). Microscopically there is extensive Zenker's degeneration (Figs. 6-8) and parasitism of the heart muscle fibers, marked cellular infiltration of the interstitial connective tissue, which are found in the ordinary musculature of every chamber of the heart (Figs. 10-11) as well as in Tawara's node (Fig. 9), main bundle (Fig. 2) and right (Fig. 4) and left (Fig. 5) septal divisions of the bundle of His, and perineuritis. Those anatomic changes are associated to an abnormal electrocardiogram presenting some similitude to that of an anemic infarct of the anterior wall of the heart and which will be discussed elsewhere (unpublished paper by Dias, Nobrega & Laranja).

Toxoplasmose congênita

We had the opportunity to study 6 cases of the congenital form of toxoplasmosis, found in a series of 1200 necropsies of fetuses and newborn babies, realized at 3 different hospitals in Rio de Janeiro, Brazil. Among the 6 cases, 4 were premature babies liveborn at the 6th-8th gestational month and 2 were stillborn (1 premature and 1 at term). In all those cases, the diagnosis was based in the detection of the parasite in tissues and in one case it was even isolated the Toxoplasma from the necrotic material found in the cranial cavity. This strain of Toxoplasma, pathogenic to pigeons, to guinea pigs and to mice, is preserved by successive transfers in mice. Some facts observed in those cases present an interest not only strictly anatomic but also have certain value for the better acknowlegment of the disease. First, we want to call the attention to the presence of a sudden high fever, during or just before pregnancy in the 4 cases in which the maternal anamnesis was perfectly studied; this fever that was preceded by a normal beginning of pregnancy, had relatively rapid remission, but in 2 cases was immediately followed by uterine bleeding and premature delivery, although the puerperium had been apparently normal. It is known that are normal the subsequent children of the mothers that delivered a baby with toxoplasmosis and that several women have normal babies before the toxoplasmotic one. We believe that the fever observed in our cases could be indicative of the beginning of maternal infection and those are the reasons why we emphasize the need of careful anamnesis, specially in the cases actually diagnosed as inapparent infection. Another fact to notice is that in 5 of our cases the event premature delivery happened always between the 6th and the 8th months of pregnancy, and the only term fetus was delivered in advanced stage of maceration. The above mentioned facts could agree with the opinion of FRENKEL (1949), when he declared that "primary infection of the pregnant mother appears more likely to be the commoner mode of fetal toxoplasmic infection", but they would disagree with WEINMAN (1952) who believes that the transmission of Toxoplasma to the fetus is more frequent through a pregnant woman with chronic disease and who says "that infection contracted during pregnancy may and probably does happen from time to time"...Still in connection with the transmission of toxoplasmosis, we want to note the verification of inflammatory lesions in the placental villi and in the umbilical cord in 3 of the 4 cases in which such organs were examined at the microscope. In the case n. 1, we found several pseudocysts of Toxoplasma in the placenta, and the fibroblasts of Wharton's jelly were particularly rich in isolated forms and in colonies of Toxoplasma; the easy multiplication of the parasite in that tissue calls the attention and even suggests its utilisation for Toxoplasma's cultivation. The confirmation of Toxoplasma in human placenta was made only recently by CRISTEN et al. (1951) and by NEGHME et al. (1952), in Chile; it is not frequent in the literature, what gives some value to our present verification. Another observation was that provided by the case n. 6. This baby, a premature one of the 6th month, was 14 days old and-died with signs of respiratory disease, the causa mortis have been pneumonia. At the necropsy, we found no gross change that suggested toxoplasmosis, except the presence of some small necrotic focuses in the cerebral nervous substance around the ventricles. As a matter of fact, there was no enlargement of spleen or liver and neither leptomeningitis nor hydrocephalus. Such focuses were attributed to possible anoxia and in fact they are extremely similar to anoxial softenings, even when they are examined at the microscope; its structure composed of a central necrotic zone, surrounded by proliferated neuroglia and by a variable deposit of calcium salts, closely simulated the anoxial softenings, when the microscopical examination is based in the common histological preparations (hematoxilin-eosin, etc.). But when we examine preparations by the Giemsa or by the periodic acid-Schiff methods, we will note the presence of Toxoplasma, with its typical aspect or a little changed by degeneration. When we describe this observation, we wish to evidence the need of the search of Toxoplasma and closed parasites, in the cases of supposed pure anoxial softenings of nervous substance, in children. The frequency with which the congenital toxoplasmosis was anatomically verified should be emphasized, although the disease had not been clinically suspected, and it should be borne in mind that the second case of toxoplasmosis reported in the world was observed in Brazil by MAGARINOS TORRES; this case was the first to be described of the generalized congenital form of the infection, i. e. with myocardial lesions and parasites in skeletal muscles and skin.

Toxoplasmose em primatas não humanos: I - Infecções naturais em Macacca Mulatta e Cebus Apella

Neste trabalho são descritas infecções naturais por Toxoplasma em um Macacca mulatta e em um Cebus apella, ambos tendo apresentado curto período de doença. Ambos se infectaram no cativeiro: o primeiro no biotério do laboratório, onde havia outros mamíferos com toxoplasmose experimental, e o segundo, na casa dos seus donos, em Jacarepaguá, on se habituara a comer carne crua como parte da alimentação. Nessa casa viviam outros platirrinos, entre os quais 2 Cebus libidinosus (um casal), os quais apresentaram Reação de Sabin-Feldman positiva (1:64), do mesmo modo que o tratador dos animais. Microscòpicamente, observaram-se nos 2 animais mortos de toxoplasmose espontânea lesões necróticas no fígado e baço, com a presença de toxoplasmas livres e intracelulares, isolados ou em associação, com maior ou menor número de indivíduos. No Cebus, no qual foi feita autópsia completa, foram observadas lesões e parasitos na suprarenal; e lesões moderadas no encéfalo, mas sem parasitos. É feita uma revisão das infecções naturais por Toxoplasma em primatas não humanos. Em mais de 60 anos de estudos do Txoplasma, foram descritas infecções naturais em apenas 31 exemplares de 18 espécies: 3 Prosimii, 10 Platyrrhinus e 5 Catharrhinus. A baixa freqüência da toxoplasmose no símios em geral é relacionada aos seus hábitos alimentares vegetarianos e insetífagos e à ecologia arborícola. A maior incidência nos platirrinos é relacionada à sua mais fácil domesticação e conseqüente mudança dos hábitos alimentares.

Toxoplasmose em primatas não humanos: II - Tentativas de infecções experimentais em Macacca mulatta, Cebus apella e Callithrix jacchus; e pesquisa de anticorpos em várias espécies de platyrrhinus

Após uma revisão da toxoplasmose experimental em primatas não humanos, são relatadas as tentativas, sem êxito, para provocar toxoplasmose aguda e fatal em dois rhesus (Macacca mulatta), um infante e outro jovem, por inoculação e reinoculação de uma amostra humana, usando diferentes vias e doses maciças e ainda com a ministração de decametasona. Do mesmo modo, não teve sucesso a tentativa para induzir a doença fatal em um Cebus apella adulto, pela via peritoneal. Porém a toxoplasmose-infecção nesses 3 animais, foi comprovada pela elevação da temperatura (39 a 41ºC), pela positividade da reação de Sabin-Feldman (1:64 - 1:256) e pelo isolamento de toxoplasmas em camundongos inoculados com material do Cebus. Por outro lado, em um Callithrix jacchus pela inoculação peritoneal, foi provocada doença grave e fatal com focos necróticos e abundãncia de toxoplasmas no baço e fígado, e isolamento dos parasitas em camundongo. De 54 símios do Nõvo Mundo, submetidos a RSF, todos foram negativos, com exceção de um Saimiri que se mostrou positivo a 1:16 (18%). Uma análise do problema Toxoplasmose-Primatas não humanos, com o apoio na revisão da literatura e nas nossas próprias observações (ver também o trabalho anterior) permite as seguintes conclusões: em seu habitat natural os primatas não humanos não são expostos ao Toxoplasma. Isso deve estar relacionado aos hábitos arborícolas e à sua alimentação vegetariana e insetívora; b) os casos descritos de toxoplasmose natural nos símios se referem a animais de cativeiro; e, mesmo nestas condições, é excepcional a infecção espontânea dos catarrinos; c) os catarrinos apresentam, além disso uma grande resistência à indução da toxoplasmose experimental, a qual não é devida à presença de anticorpos circulantes. Essa resistência parece não ser rompida pela administração de corticosteróides; embora às vêzes o seja pela inoculação de doses maciças de toxoplasmas, e geralmente nos animais jovens; d) essa resistência é menor nos platirrinos e parece não existir nos platirrinos inferiores e nos Prosimii.

Indirect immunofluorescence(IgG and IgM) tests for toxoplasmosis on 203 persons, with no symptomatology suggesting the disease, located in the city of Rio de Janeiro. Serological follow up one to two years later.

Clinical and serological follow up examinations were performed on 203 persons, from three to twenty years of age, from the otolaryngology department of a hospital in the city of Rio de Janeiro, with no symptomatology suggesting toxoplasmosis, but suffering from chronic tonsillitis. According to results obtained during the first indirect immunofluorescence tests, the patients were divided into following groups: Group I (non-reactive IgG and IgM), 98 persons (48.3%); Group II (1:16 ≤ IgG ≤ 1:256 and non-reactive IgM), 74 persons (36.5%); Group III (IgM ≥ 1:1024 and non-reactive IgM), 18 persons (8.8%), and Group IV (IgG and IgM reactive), 13 persons (6.4%). One to two years later, 131 (64.5%) of the 203 persons were reexamined by a second indirect immunofluorescence test. In the case of 66 persons (Group I) whose serum was non-reactive in the IgG and IgM classes during the first indirect immunofluorescence test, serum conversion was observed in aproximately 21.2%. in 65 individuals (49.6%), (Groups II, III and IV),with reactive serum in the IgG classes during the first indirect immunofluorescence test, the second reaction showed an increase in titres in 20% of the cases, a decrease in 67.7% of the cases, or no alterations in 12.3 of the cases. In the IgM class, all 131 sera were non-reactive at 116 dilution the second immunofluorescence test, including the 13 cases that had previously been reactive in the immunoglobulin class, Symptomatology suggesting toxoplasmosis was only observed in one case during the second testing, this patient's principal physical sign being hypertrophied lymph nodes. during this period, the Toxoplasma antibodies showed titres of IgG 1:32000 and non-reactive IgM, whilst one year previously, during the first test, these titres were IgG 1:1024 and IgM 1:64. Differences in the age, sex and skin coloring of patients were not statistically significant as regards alterations in the indirect immunofluorescence test titres.

Toxoplasmose humana adquirida através da ingestão de leite de cabra

A ocorrência de toxoplasmose aguda em três membros de uma mesma família foi relacionada à ingestão de leite de cabra, não pasteurizado e nem fervido. As cabras eram criadas soltas no peridomicílio para fornecimento de leite. Anticorpos fluorescentes anti-Toxoplasma foram detectados nos quinze animais examinados, sendo que oito caprinos apresentaram títulos superiores a 1:1024 e nas cinco cabras lactantes estes títulos variaram de 1:1024 a 1:32.768. Taquizoitos foram isolados, por inovulação em camundongos, do leite de uma destas cabras. Os cães criados na mesma casa não apresentaram sintomas de toxoplasmose apesar de ter ocorrido perda de ninhada de uma cadela; em todos os cães foram detectados baixos níveis de anticorpos. Foi considerada menos provável a possibilidade de que as infecções humanas pudessem ser devido à ingestão de alimentos contaminados com oocistos.

Molecular approaches to malaria and Babesisosis diagnosis

The development of additional methods for detecting and identifuing Babesia and Plasmodium infections may be useful in disease monitoring, management and control efforts. To preliminarily evaluate sunthetic peptide-based serodiagnosis, a hydrophilic sequence (DDESEFDKEK)was selected from published BabR gene of B. bovis. Immunization of rabbits and cattle with the hemocyanin-conjugated peptide elicited antibody responses that specifically detected both P. falciparum and B. bovis antigens by immunofluorescence and Western blots. Using a dot-ELISA with this peptide, antisera from immunized and naturally-infected cattle, and immunized rodents, were specifically detected. Reactivity was weak and correlated with peptide immunization or infection. DNA-based detection using repetitive DNA was species-specific in dot-blot formats for B. bovis DNA, and in both dot-blot and in situ formats for P. falciparum; a streamlined enzymelinked synthetic DNA assay for P. falciparum detected 30 parasites/mm(cúbicos) from patient blood using either colorimetric (2-15 h color development) or chemiluminescent detection (0.5-6-min. exposures). Serodiagnostic and DNA hybridization methods may be complementary in the respective detection of both chronic and acute infections. However, recent improvements in the polymerase chain reaction (PCR) make feasible a more sensitive and uniform approach to the diagnosis of these and other infectious disease complexes, with appropriate primers and processing methods. An analysis of ribosomal DNA genes of Plasmodium and Toxoplasma identified Apicomplexa-conserved sequence regions. Specific and distinctive PCR profiles were obtained for primers spanning the internal transcribed spacer locus for each of several Plasmodium and Babesia species.

Role of the immune response induced by superantigens in the pathogenesis of microbial infections.

Superantigens (SAgs) are microbial proteins which have potent effects on the immune system. They are presented by major histocompatibility complex (MHC) class II molecules and interact with a large number of T cells expressing specific T cell receptor V beta domains. Encounter of a SAg leads initially to the stimulation and subsequently to the clonal deletion of reactive T cells. SAgs are expressed by a wide variety of microorganisms which use them to exploit the immune system to their own advantage. Bacterial SAgs are exotoxins which are linked to several diseases in humans and animals. A classical example is the toxic shock syndrome in which the massive release of cytokines by SAg-reactive cells is thought to play a major pathogenic role. The best characterized viral SAg is encoded by mouse mammary tumour virus (MMTV) and has proved to have a major influence on the viral life cycle by dramatically increasing the efficiency of viral infection. In this paper, we review the general properties of SAgs and discuss the different types of microorganisms which produce these molecules, with a particular emphasis on the role played by the SAg-induced immune response in the course of microbial infections.

Parvovirus B19 infections in state of Rio de Janeiro, Brasil: 526 sera analyzed by IgM-enzyme-linked immunosorbent assay and polymerase chain reaction

In this study were analyzed 526 sera; the patients aged from two days to 65 years old presenting exanthema, which was the most frequent symptom observed, besides fever, adenomegaly, and arthralgia. These sera were negative by enzyme-linked immunosorbent assay (IgM-ELISA) for either rubella (495), toxoplasma (41), cytomegalovirus (12), measles (40), dengue (56), and they were submitted to nested polymerase chain reaction (PCR) for B19 DNA and commercial IgM-ELISA for B19. In 39 abortion cases, IgM or DNA were not detected, therefore they were not took into account for analysis. Specific DNA and IgM were detected respectively in 71 (14.5%) and IgM in 62 (12.7%) sera from 487 sera analyzed. IgM and DNA were simultaneously detected in 43 (8.8%), while agreement among the results by PCR and IgM-ELISA was observed in 440 (90.4%). The sera were collected from January 1999 to December 2000, most of them in 1999 (325), during winter and spring. The major number of clinical cases was observed in the age group from one to ten years old. IgM or DNA were detected in 23 from 51 municipal districts of the state of Rio de Janeiro, where the samples were collected.

The Complete Chromosomal Organization of the Reference Strain of the Leishmania Genome Project, L. major ;Friedlin'.

In recent years, analysis of the genomes of many organisms has received increasing international attention. The bulk of the effort to date has centred on the Human Genome Project and analysis of model organisms such as yeast, Drosophila and Caenorhabditis elegans. More recently, the revolution in genome sequencing and gene identification has begun to impact on infectious disease organisms. Initially, much of the effort was concentrated on prokaryotes, but small eukaryotic genomes, including the protozoan parasites Plasmodium, Toxoplasma and trypanosomatids (Leishmania, Trypanosoma brucei and T. cruzi), as well as some multicellular organisms, such as Brugia and Schistosoma, are benefiting from the technological advances of the genome era. These advances promise a radical new approach to the development of novel diagnostic tools, chemotherapeutic targets and vaccines for infectious disease organisms, as well as to the more detailed analysis of cell biology and function.Several networks or consortia linking laboratories around the world have been established to support these parasite genome projects[1] (for more information, see http://www.ebi.ac.uk/ parasites/paratable.html). Five of these networks were supported by an initiative launched in 1994 by the Specific Programme for Research and Tropical Diseases (TDR) of the WHO[2, 3, 4, 5, 6]. The Leishmania Genome Network (LGN) is one of these[3]. Its activities are reported at http://www.ebi.ac.uk/parasites/leish.html, and its current aim is to map and sequence the genome of Leishmania by the year 2002. All the mapping, hybridization and sequence data are also publicly available from LeishDB, an AceDB-based genome database (http://www.ebi.ac.uk/parasites/LGN/leissssoft.html).

Oocyst wall formation and composition in coccidian parasites

The oocyst wall of coccidian parasites is a robust structure that is resistant to a variety of environmental and chemical insults. This resilience allows oocysts to survive for long periods, facilitating transmission from host to host. The wall is bilayered and is formed by the sequential release of the contents of two specialized organelles - wall forming body 1 and wall forming body 2 - found in the macrogametocyte stage of Coccidia. The oocyst wall is over 90% protein but few of these proteins have been studied. One group is cysteine-rich and may be presumed to crosslink via disulphide bridges, though this is yet to be investigated. Another group of wall proteins is rich in tyrosine. These proteins, which range in size from 8-31 kDa, are derived from larger precursors of 56 and 82 kDa found in the wall forming bodies. Proteases may catalyze processing of the precursors into tyrosine-rich peptides, which are then oxidatively crosslinked in a reaction catalyzed by peroxidases. In support of this hypothesis, the oocyst wall has high levels of dityrosine bonds. These dityrosine crosslinked proteins may provide a structural matrix for assembly of the oocyst wall and contribute to its resilience.