Analysis of cytokines IFN-γ, TNF-α, TGF-ß and nitric oxide in amniotic fluid and serum of pregnant women with toxoplasmosis in southern Brazil.

This study detected and compared the levels of IFN-γ, TNF-α, TGF-ß and nitric oxide (NO) in amniotic fluid (AF) and serum of pregnancies with acute toxoplasmosis, Southern Brazil. It also was compared the levels of the same mediators in the serum of pregnancies in acute and chronic toxoplasmosis with non-infected. Serological investigation, anti-T gondii IgM and IgG, of the 67 pregnancies was determined by Elisa MEIA. Forty two were uninfected, eight in chronic phase and 17 in acute phase. Among the acute phase, seven agreed to amniocentesis. The cytokines, in serum and in AF, were assessed by sandwich ELISA, and NO was estimated from the nitrite measurement with Griess reagent. The IFN-γ and TGF-ß levels in the AF and blood were similar, while TNF-α levels was lower in the AF. On the other hand, NO was higher in the AF. Chronically infected pregnant women have showed lower levels of INF-γ than those in acute and uninfected pregnancies. The serological levels of TNF-α were lower in pregnancies with toxoplasmosis, when compared with non-infected. TGF-ß levels were higher in pregnancies in acute phase when compared with uninfected or chronically infected. NO in the serum of the infected had lower levels than those non-infected. In summary, higher concentrations of NO and lower levels of TNF-α were observed in the AF than in the serum of acute pregnancies, while TGF-ß e INF-γ levels were similar in both biological material. In the serum of infected pregnancies was observed decrease in inflammatory mediators and increase of TGF-ß.

An alternative method to recover Toxoplasma gondii from greenery and fruits.

Toxoplasma gondii oocysts are an important form of contamination with a high dispersion in the environment, but their detection is still a challenge. This study evaluated the recovery of oocysts from strawberries and crisphead lettuce. Samples (250 g of strawberries or one head of lettuce) were experimentally inoculated with 10, 10(2), 10(3) and 10(4) T. gondii oocysts, by two separate processes, spot dripping and immersion. Then, 50 g of each sample was washed, filtered through a cellulose ester membrane, and concentrated by centrifugation. Three aliquots were taken for DNA extraction in a direct way, after freeze-thaw (FT) cycles or ultrasound (US), followed by PCR (B22-B23 and Tox4-Tox5 primers). The T. gondii DNA was amplified with the primers B22-B23 in all samples contaminated by dripping and when DNA extraction was carried out after FT or US. These techniques may be useful in epidemiological surveillance in the control of this zoonosis.

Predominance of Giardia duodenalis Assemblage AII in Fresh Leafy Vegetables from a Market in Southern Brazil.

We investigated the presence of Giardia duodenalis cysts and its genotypes in raw leafy vegetables sold in a Brazilian market. These products are different from those sold in most street markets because the producers themselves display and sell their products and rely on specialized technical and sanitary assistance. Vegetable and water samples were collected from 14 (80%) producers who cultivated vegetables that are typically consumed raw for sale at the market, obtained at the market and farms, respectively. A total of 128 samples of leafy greens (chives, parsley, cabbage, arugula, watercress, and chicory) and 14 water samples were analyzed by direct immunofluorescence and PCR techniques. The positive samples were genotyped (GHD gene) using PCR and restriction fragment length polymorphism. The analyses indicated that 16 (12.5%) of 128 samples were positive by PCR, while 1 (0.8%) of 128 samples were positive by immunofluorescence. Giardia cysts were not detected in the water samples obtained at the farms. The molecular technique revealed a genotype with zoonotic potential, which underscores the challenge in the control of giardiasis dissemination via the consumption of raw vegetables.

WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

Aborto espontâneo e toxoplasmose ocular em um casal infectado com Toxoplasma gondii / Spontaneous abortion and ocular toxoplasmosis in a couple infected with Toxoplasma gondii

OBJETIVOS: Relatar os casos de um casal, em que a esposa apresentou toxoplasmose na gestação e o esposo, após 15 anos, foi diagnosticado com toxoplasmose ocular. RELATO DOS CASOS: Gestante, grávida de gêmeos, residente de um estado na região Sul do Brasil. No primeiro mês de gestação apresentou sorologia não reagente para IgG e IgM anti-Toxoplasma gondii. Na sétima semana de gestação houve soroconversão (IgM e IgG anti-T.gondii reagentes) e aborto natural. O esposo, após 15 anos desde o episódio da esposa, queixou-se de visão embaçada/irritada. O exame de fundoscopia revelou lesões típicas de coriorretinite recidivante por toxoplasmose no olho direito. Os exames sorológicos foram IgG anti-T.gondii reagente e IgM anti-T.gondii não reagente. CONCLUSÕES: Este relato, de um casal com desfechos clínicos importantes de toxoplasmose, em diferentes formas, enfatiza a relevância do problema da toxoplasmose, que nas suas diversas manifestações, pode trazer graves consequências para a vida de uma família.

AIMS: To report the cases of wife and husband with toxoplasmosis. The woman acquired the infection during pregnancy while her husband was diagnosed with ocular toxoplasmosis 15 years later. CASES REPORT: A woman with twin pregnancy, living in southern Brazil, was seronegative for Toxoplasma gondii IgG and IgM antibodies in the first month of gestation. Seroconversion (positive T. gondii IgM and IgG antibodies) occurred in the seventh week of gestation, resulting in spontaneous abortion. Fifteen years later, her husband presented with eye irritation and blurred vision. The funduscopic examination revealed typical lesions of recurrent chorioretinitis in the right eye. The serological tests were positive for T. gondii IgG and negative for IgM. CONCLUSIONS: This report of a couple with important clinical outcomes of two different types of toxoplasmosis underscores the importance of this disease which, in its various manifestations, may have serious consequences for a family's life.