Sheep abortion associated with Neospora caninum in Mato Grosso do Sul, Brazil

Canids are the main hosts of Neospora caninum, but cattle, (sheep, goats and horses may serve as intermediary hosts. N. caninum infection of pregnant intermediary hosts may provoke abortion and neonatal infections. This study is the first to report lamb abortion associated with N. caninum in Mato Grosso do Sul. Epidemiological data were obtained from interviews with sheep producers. For microscopic examination, fragments of different organs removed from 4 sheep fetuses, aborted and necropsied, were fixed in 10% formaldehyde, embedded in paraffin and subjected to the hematoxylin-eosin staining protocol and immunohistochemistry (IHC) to test for N. caninum and Toxoplasma gondii. The abortion outbreak studied was reported from a herd of 268 Santa Inês sheep (including 186 pregnant ewes), with 10 abortion cases in the last third of gestation. Four fetuses were examined, 3 from a same ewe. At necropsy, one fetus exhibited crackling in the lung and all its organs were reddish. Histological findings detected mononuclear cell infiltrates among myocardium fibers and around blood vessels, in addition to circular structures with basophilic points resembling protozoans. IHC tests revealed strongly positive staining for N. caninum and weakly positive for T. gondii, characterizing N. caninum infection.

Toxoplasmose em primatas neotropicais: estudo retrospectivo de sete casos

A toxoplasmose é considerada uma doença parasitária fatal em primatas neotropicais. O objetivo deste trabalho foi descrever, através de um estudo retrospectivo, os casos de toxoplasmose em primatas neotropicais. No período de 1999-2009 foram realizados 86 exames anatomopatológicos em primatas e a toxoplasmose foi a enfermidade mais comum (7/86), relatando-se um caso em sagui-do-tufo-preto (Callithrix penicillata) e seis em bugio-ruivo (Alouatta guariba). Dois animais foram encontrados mortos e cinco morreram em poucos dias. Os sinais clínicos mais frequentes foram apatia e anorexia (5/7), distensão abdominal (4/7) e febre (3/7). Na necropsia observou-se esplenomegalia (4/7), hemorragia do trato digestório, linfonodos e bexiga (4/7), pulmões avermelhados (3/7) e hepatomegalia (2/7). No exame histopatológico evidenciou-se hepatite (7/7), esplenite (3/7), miocardite (2/7), enterite (2/7), linfadenite (1/7) e sialite (1/7) necróticas e, pneumonia intersticial (4/7). Em fígado, pulmões, baço, coração, linfonodos e glândula salivar havia taquizoítos de Toxoplasma gondii que foram também detectados pelo exame de imuno-histoquímica anti-T. gondii em fígado, baço e pulmões (5/7). A toxoplasmose pode causar alta mortalidade em colônias de primatas neotropicais e representar mais uma ameaça à conversação dessas espécies em cativeiro. Sendo assim, medidas preventivas devem ser tomadas para evitar a contaminação desses animais.

Induction of cell-mediated immunity during early stages of infection with intracellular protozoa

Toxoplasma gondii and Trypanosoma cruzi are intracellular parasites which, as part of their life cycle, induce a potent cell-mediated immunity (CMI) maintained by Th1 lymphocytes and IFN-g. In both cases, induction of a strong CMI is thought to protect the host against rapid parasite multiplication and consequent pathology and lethality during the acute phase of infection. However, the parasitic infection is not eliminated by the immune system and the vertebrate host serves as a parasite reservoir. In contrast, Leishmania sp, which is a slow growing parasite, appears to evade induction of CMI during early stages of infection as a strategy for surviving in a hostile environment (i.e., inside the macrophages which are their obligatory niche in the vertebrate host). Recent reports show that the initiation of IL-12 synthesis by macrophages during these parasitic infections is a key event in regulating CMI and disease outcome. The studies reviewed here indicate that activation/inhibition of distinct signaling pathways and certain macrophage functions by intracellular protozoa are important events in inducing/modulating the immune response of their vertebrate hosts, allowing parasite and host survival and therefore maintaining parasite life cycles.

The use of confocal laser scanning microscopy to analyze the process of parasitic protozoon-host cell interaction

In this communication we review the results obtained with the confocal laser scanning microscope to characterize the interaction of epimastigote and trypomastigote forms of Trypanosoma cruzi and tachyzoites of Toxoplasma gondii with host cells. Early events of the interaction process were studied by the simultaneous localization of sites of protein phosphorylation, revealed by immunocytochemistry, and sites of actin assembly, revealed by the use of labeled phaloidin. The results obtained show that proteins localized in the interaction sites are phosphorylated. The process of formation of the parasitophorous vacuole was monitored by labeling the host cell surface with fluorescent probes for lipids (PKH26), proteins (DTAF) and sialic acid (FITC-thiosemicarbazide) before interaction with the parasites. Evidence was obtained indicating transfer of components of the host cell surface to the parasite surface in the beginning of the interaction process. We also analyzed the distribution of cytoskeletal structures (microtubules and microfilaments visualized with specific antibodies), mitochondria (visualized with rhodamine 123), the Golgi complex (visualized with C6-NBD-ceramide) and the endoplasmic reticulum (visualized with anti-reticulin antibodies and DIOC6) during the evolution of intracellular parasitism. The results obtained show that some, but not all, structures change their position during evolution of the intracellular parasitism.

Enucleated L929 mouse fibroblasts support invasion and multiplication of Shigella flexneri 5a

Invasive bacteria can induce their own uptake and specify their intracellular localization; hence it is commonly assumed that proximate modulation of host cell transcription is not required for infection. However, bacteria can also modulate, directly or indirectly, the transcription of many host cell genes, whose role in the infection may be difficult to determine by global gene expression. Is the host cell nucleus proximately required for intracellular infection and, if so, for which pathogens and at what stages of infection? Enucleated cells were previously infected with Toxoplasma gondii, Chlamydia psittaci, C. trachomatis, or Rickettsia prowazekii. We enucleated L929 mouse fibroblasts by centrifugation in the presence of cytochalasin B, and compared the infection with Shigella flexneri M90T 5a of nucleated and enucleated cells. Percent infection and bacterial loads were estimated with a gentamicin suppression assay in cultures fixed and stained at different times after infection. Enucleation reduced by about half the percent of infected cells, a finding that may reflect the reduced endocytic ability of L929 cytoplasts. However, average numbers of bacteria and frequency distributions of bacterial numbers per cell at different times were similar in enucleated and nucleated cells. Bacteria with actin-rich tails were detected in both cytoplasts and nucleated cells. Lastly, cytoplasts were similarly infected 2 and 24 h after enucleation, suggesting that short-lived mRNAs were not involved in the infection. Productive S. flexneri infection could thus take place in cells unable to modulate gene transcription, RNA processing, or nucleus-dependent signaling cascades.

Lectins for the detection of IgM antibodies to T. gondii in the diagnosis of acute toxoplasmosis by immunofluorescence test

Lectins were labeled with fluorescein and tried as conjugates in the immunofluorescence (IP) test for the detection of IgM antibodies to T. gondii, in the diagnosis of acute toxoplasmosis. This approach was an attempt to find alternative reagents for anti-human IgM fluorescent conjugates (AHIgMFC), which contain quite frequently anaibcdies to toxoplasma, as contaminants, due to natural T. gondii infections among animals used for imunization. Lentil (Lens culinaris) lectin fluorescence conjugates (LcFC) provided most satisfactory results. The evaluation of LcFC carried out in a total of 179 sera from patients with acute and chronic toxoplasmosis, with non-related infections or healthy subjects, gave high values of relative efficiency, co-positivity and co-negativity indices, respectively 0.989, 0.969 and 1.000, in reference to the conventional AHIgMFC. Moreover, three batches of LcFC successively prepared gave reproducible test results. The advantage of LcFC as an alternative reagent for the serodiagnosis of acute toxoplasmosis is supported by practical aspects of its preparation.

High diagnostic efficiency of IgM-ELISA with the use of multiple antigen peptides (MAP1) from T. gondii ESA (SAG-1, GRA-1 and GRA-7), in acute toxoplasmosis

The main serological marker for the diagnosis of recent toxoplasmosis is the specific IgM antibody, along with IgG antibodies of low avidity. However, in some patients these antibodies may persist long after the acute/recent phase, contributing to misdiagnosis in suspected cases of toxoplasmosis. In the present study, the diagnostic efficiency of ELISA was evaluated, with the use of peptides derived from T. gondii ESA antigens, named SAG-1, GRA-1 and GRA-7. In the assay referred to, we studied each of these peptides individually, as well as in four different combinations, as Multiple Antigen Peptides (MAP), aiming to establish a reliable profile for the acute/recent toxoplasmosis with only one patient serum sample. The diagnostic performance of the assay using MAP1, with the combination of SAG-1, GRA-1 and GRA-7 peptides, demonstrated better discrimination of the acute/recent phase from non acute/recent phase of toxoplasmosis. Our results show that IgM antibodies to MAP1 may be useful as a serological marker, enhancing the diagnostic efficiency of the assay for acute/recent phase of toxoplasmosis.

Detection of anti-Giardia lamblia serum antibody among children of day care centers

OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA) for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3%) showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3%) and 100 (68%) samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39%) than IIF (70%). IIF also showed a higher concordance with microscopic examination than ELISA.

Acute Hepatitis B in a patient previously positive for antibody to surface antigen (anti-HBs) determined by radioimmunoassay: case report and review of the literature

The determination of anti-HBs as a screening test before vaccination has been advisable in order to encounter immune individuals that don't need to receive vaccine protection. A case-report is presented and three other cases are reviewed from the literature. Anti-HBs was positive in these health-care personnels that developped typical acute B hepatitis. Different subtyping involving the d/y determinants were found in the first case, but false-positive anti-HBs even with high titres, determined by RIA, were found in the other cases. Concomitant determination of anti-HBc or absence of screening tests seem to be more reasonable policies until a low-cost and risk-free vaccine is produced.

Prevalence of antibody to hepatitis B virus in an urban population of northeast Brazil

A sample of 1,288 inhabitants of Salvador, Bahia, Brazil, were submitted to the determination of anti-HBs using radioimmunoassay procedure, and analysed according to age, sex and income. Overall prevalence of anti-HBs was 11,8%. ranging from 6,7% among children aged less than three years old to 26,1% among those aged 30 years and older. Males presented prevalence of anti-HBs similar to female individuals, and those with a higher income showed frequencies of anti-HBs greater than those with a lower income level. The following conclusions were drawn: The high prevalence of anti-HBs observed among children suggests early contact with hepatitis B virus, possibly due to vertical transmission and intrafamiliar dissemination of the disease; the frequency of anti-HBs increases with age; the lower prevalence of anti-HBs among those with low income suggests that this group may present higher prevalence of carriers of the hepatitis B virus surface antigen.

Antibody response to heat-inactivated hepatitis B vaccine (CLB-3mg) in hemodialysis patients and occupational risk personnel: a one year follow-up

Immune response against hepatitis B vaccine (CLB 3mg) was evaluated in 59 hemodialysis patients and 20 occupational risk personnel. Seroconversion was induced in 52.5% and 70.0% respectively. Twelve months after the first dose, 37.5% of patients and 60.0% of occupational risk personnel had detectable anti-HBs level. Antibody level was expressed in sample ratio units (SRU). Considering only the responders, in the patients group 38.7% had a low anti-HBs response (2.1-9.9 SRU) 32.3% a medium response (10-99.9 SRU) and 29.0% a high response (>100 SRU) while in occupational risk personnel these values were 14.3%, 64.3% and 21.4% respectively. The authors suggest the use of HBV vaccines with more elevated HBsAg concentration or a reinforced immunization schedule to improve the anti-HBs response not only for patients but also for healthy persons.

Immunofluorescent antibody to rotavirus and antigen excretion in infantile acute diarrhoea

A serological study was carried out to evaluate the causal relationship of acute diarrhoea and rotavirus in children under five years of age. The rotaviral infection was demonstrated in paired sera by determining the antibody titers by immunofluorescence test (IF). Bovine rotavirus-infected MA-104 cell culture was used as substratum for IF. Out of 80 paired sera it was shown that 23 (28.75%) presented seroconversion, 19 (23.75%) samples showed a twofold increase in their titers and 38 (47.5%) had no increase in rotavirus antibody. This result is discussed on the light of previously obtained results on viral antigen detection by counterimmunoelectrosmophoresis (CIEOP).

Peroxidase antibody test for mucocutaneous leishmaniasis serology performance indexes and comparison with a fluorescent antibody test: Comparação com o desempenho da reação de imunofluorescência

Performance indexes of the peroxidase antibody test were compared to that of the fluorescent antibody test. The peroxidase antibody test had a statistically higher sensitivity and negative predictive value and a higher efficiency than the fluorescent antibody test but its specificity and positive predictive value were within the 95% confidence limits for the values found for the fluorescent antibody test. Such differences did not change when Chagas' disease and visceral leishmaniasis sera were included in index calculations. Statistical analysis showed that the two tests have a substantial degree of agreement but the immunofluorescent test had a specificity index and a positive predictive value equal to 100.0% when Chagas' disease and visceral leishmaniasis sera were not included in the calculations of the performance index; in this instance, a positive test result equals a disclosure of the disease attribute due to the inexistence of false positive results. The enzyme/ protein ratio of the peroxidase conjugate, resulting in heavy or light-labeled conjugates may pose technical problems to its use in serology tests.

The prevalence of antibody to human parvovirus B19 in Rio de Janeiro, Brazil

During 1985 and 1986 serum samples were collected from the Rio de Janeiro population and examined for the presence of IgG antibody to human parvovirus B19. No difference in prevalence was found between males and females. Antibody prevalence rose from 35% in children less than five years old to almost 80% in children aged eleven to fifteen years. The antibody prevalence in individuals over 50 years old was over 90%.