Thesis (Ph.D.). / Written for the Dept. of Natural Resource Sciences, Macdonald College of McGill University. Title from title page of PDF (viewed 2008/08/04). Includes bibliographical references.

Thesis (M.S.)--University of Wisconsin--Stevens Point, 2002. / Includes bibliographical references (leaves 57-63).

Neste trabalho, foram desenvolvidos e validados métodos analítico e microbiológico, bem como estudo preliminar de estabilidade, cinética de degradação e citotoxicidade da Ceftarolina Fosamila em pó para solução injetável, um antibiótico da classe das cefalosporinas de quinta geração, indicado para pneumonias adquiridas na comunidade e infecções graves, de pele e tecidos moles. A validação do ensaio microbiológico pelo método de difusão em ágar cilindros em placa, delineamento 3x3, apresentou resultados satisfatórios, como especificidade, linearidade na faixa de 2,0 - 8,0 μg/mL, precisão (109,42 %), exatidão (102,3 %) e robustez. Soluções de Cefatarolina Fosamila do produto acabado expostas à radiação UVC (254 nm) e à degradação térmica a 60 °C foram utilizadas para avaliar a especificidade do bioensaio. A robustez foi avaliada através da alteração da concentração do meio inoculado (0,8 e 1,2 %). O desenvolvimento e validação de método por CLAE foi avaliado através da especificidade, linearidade, precisão, exatidão e robustez. No método cromatográfico foi utilizado cromatógrafo à liquido de alta eficiência SHIMADZU com coluna Agilent® C18, fase móvel (água com trietilamina 1,0% pH 5,0:acetonitrila 87:13 v/v). O método apresentou-se específico, linear, no intervalo de 5,0 - 60,0 μg/mL, preciso (110,0 %), exato (100,68 %) e robusto. Os métodos microbiológico e cromatográfico validados foram comparados estatisticamente e verificou-se não haver diferença significativa entre eles quando comparados através do teste “t” de Student. No estudo preliminar de estabilidade constatou-se ser estável em hidrólise ácida (0,1 M) e luz UVA no período avaliado, e instável frente à degradação térmica (40 e 60 °C), oxidativa com peróxido de hidrogênio, básica em NaOH (0,1 M e 0,01 M) e luz UVC. As cinéticas de degradação frente à luz UVC e degradação térmica 60 °C mostraram que as amostras possuem cinética de degradação de ordem zero e de segunda ordem, respectivamente. O ensaio de citotoxicidade demonstrou não haver diferença entre a condição normal e a amostra submetida à degradação forçada, sugerindo que os possíveis produtos de degradação formados não alteraram o resultado. / In this work, analytical and microbiological methods were developed and validated, as well as a preliminary study of the stability, degradation kinetics and cytotoxicity to Ceftaroline Fosamil powder for injectable solution, this is a fifth generation cephalosporin antibiotic indicated for community-acquired pneumonia and severe infections of the skin and soft tissues. The validation of the microbial assay by diffusion method in 3x3 cylinder agar delineated showed satisfactory results in specificity, linearity in the range of 2.0 - 8.0 μg / mL, precision (109.42 %), accuracy (102.3 %) and robustness. The development and validation of the method by HPLC was evaluated through specificity, linearity, precision, accuracy and robustness. In the chromatographic method was used high performance liquid chromatograph from SHIMADZU with Agilent® C18 column, mobile phase (water with triethylamine 1.0 % pH 5.0: acetonitrile 87:13 v/v). The method was linear, specific in the range of 5.0 - 60.0 μg/mL, accurate (110.0 %), exact (100.68 %) and robust. The validated microbiological and chromatographic methods were compared statistically and there was no significant difference between them when compared through Student's t-test. In the preliminary stability study, it was found stable in acid hydrolysis (0.1M) and UVA light in the period evaluated, and instable against thermal degradation (40 and 60 °C), oxidative with hydrogen peroxide, basic in NaOH (0.1 M and 0.0 1M) and UVC light. Samples exposed in UVC light an thermal degradation at 60°C showed degradation kinetics following zero order and second order, respectively. The cytotoxicity assay showed no difference between the normal condition and the sample submitted to forced degradation, suggesting that the possible degradation products formed did not change the result.

Neste trabalho, foram desenvolvidos e validados métodos analítico e microbiológico, bem como estudo preliminar de estabilidade, cinética de degradação e citotoxicidade da Ceftarolina Fosamila em pó para solução injetável, um antibiótico da classe das cefalosporinas de quinta geração, indicado para pneumonias adquiridas na comunidade e infecções graves, de pele e tecidos moles. A validação do ensaio microbiológico pelo método de difusão em ágar cilindros em placa, delineamento 3x3, apresentou resultados satisfatórios, como especificidade, linearidade na faixa de 2,0 - 8,0 μg/mL, precisão (109,42 %), exatidão (102,3 %) e robustez. Soluções de Cefatarolina Fosamila do produto acabado expostas à radiação UVC (254 nm) e à degradação térmica a 60 °C foram utilizadas para avaliar a especificidade do bioensaio. A robustez foi avaliada através da alteração da concentração do meio inoculado (0,8 e 1,2 %). O desenvolvimento e validação de método por CLAE foi avaliado através da especificidade, linearidade, precisão, exatidão e robustez. No método cromatográfico foi utilizado cromatógrafo à liquido de alta eficiência SHIMADZU com coluna Agilent® C18, fase móvel (água com trietilamina 1,0% pH 5,0:acetonitrila 87:13 v/v). O método apresentou-se específico, linear, no intervalo de 5,0 - 60,0 μg/mL, preciso (110,0 %), exato (100,68 %) e robusto. Os métodos microbiológico e cromatográfico validados foram comparados estatisticamente e verificou-se não haver diferença significativa entre eles quando comparados através do teste “t” de Student. No estudo preliminar de estabilidade constatou-se ser estável em hidrólise ácida (0,1 M) e luz UVA no período avaliado, e instável frente à degradação térmica (40 e 60 °C), oxidativa com peróxido de hidrogênio, básica em NaOH (0,1 M e 0,01 M) e luz UVC. As cinéticas de degradação frente à luz UVC e degradação térmica 60 °C mostraram que as amostras possuem cinética de degradação de ordem zero e de segunda ordem, respectivamente. O ensaio de citotoxicidade demonstrou não haver diferença entre a condição normal e a amostra submetida à degradação forçada, sugerindo que os possíveis produtos de degradação formados não alteraram o resultado. / In this work, analytical and microbiological methods were developed and validated, as well as a preliminary study of the stability, degradation kinetics and cytotoxicity to Ceftaroline Fosamil powder for injectable solution, this is a fifth generation cephalosporin antibiotic indicated for community-acquired pneumonia and severe infections of the skin and soft tissues. The validation of the microbial assay by diffusion method in 3x3 cylinder agar delineated showed satisfactory results in specificity, linearity in the range of 2.0 - 8.0 μg / mL, precision (109.42 %), accuracy (102.3 %) and robustness. The development and validation of the method by HPLC was evaluated through specificity, linearity, precision, accuracy and robustness. In the chromatographic method was used high performance liquid chromatograph from SHIMADZU with Agilent® C18 column, mobile phase (water with triethylamine 1.0 % pH 5.0: acetonitrile 87:13 v/v). The method was linear, specific in the range of 5.0 - 60.0 μg/mL, accurate (110.0 %), exact (100.68 %) and robust. The validated microbiological and chromatographic methods were compared statistically and there was no significant difference between them when compared through Student's t-test. In the preliminary stability study, it was found stable in acid hydrolysis (0.1M) and UVA light in the period evaluated, and instable against thermal degradation (40 and 60 °C), oxidative with hydrogen peroxide, basic in NaOH (0.1 M and 0.0 1M) and UVC light. Samples exposed in UVC light an thermal degradation at 60°C showed degradation kinetics following zero order and second order, respectively. The cytotoxicity assay showed no difference between the normal condition and the sample submitted to forced degradation, suggesting that the possible degradation products formed did not change the result.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Steven S. Dritz / Cassandra K. Jones / Biological hazards in animal feed are a growing concern for the feed industry. Porcine epidemic diarrhea virus (PEDV) is the first viral pathogen confirmed to be transmissible in swine feed and feed ingredients. This led to investigations identifying the magnitude of transmissible risk PEDV imposes and strategies to mitigate infectivity in contaminated diets. The objective of the first experiment was to evaluate the minimum infectious dose of PEDV in virus-inoculated feed. Pigs became infected when PEDV concentrations at or above 5.6 × 10¹ 50% tissue culture infectious dose/g (TCID₅₀/g); corresponding feed cycle threshold (Ct) of 37 or below was utilized. Evaluation of a mitigation strategy for PEDV contaminated diets is also important since cross-contamination during feed manufacturing is possible. Therefore, the objective of the second experiment was to determine the effectiveness of feed batch sequencing as a method to minimize the risk of PEDV cross-contamination. This method was effective to reduce but not eliminate infectious PEDV carryover risk. Furthermore, feed that lacked detectible PEDV RNA as analyzed by quantitative real-time reverse transcription PCR assay (qPCR) was infectious. The third study was an observational study complementary to the previous experiment where the magnitude of virus contamination in the feed manufacturing facility was characterized during feed batch sequencing. Widespread contamination of the facility occurred and surfaces remained contaminated until chemically cleaned. The final experiment was conducted to assess PEDV RNA detection in feed and spray dried porcine plasma (SDPP) when analyzed by qPCR across 5 diagnostic laboratories. Overall, it appears qPCR PEDV RNA detection in feed and SDPP was precise as quantified by low coefficient of variation across laboratories, with the exception of one %CV from SDPP inoculated with low virus load from one laboratory. Although the magnitude of the Ct value difference was large in only 1 of 5 laboratories, comparisons of Ct values across laboratories should be interpreted cautiously. Finally, qPCR can be a useful surveillance tool for detection of PEDV RNA in non-clinical samples such as feed and SDPP.

The focus of this study is the hydrochemical variability of runoff events in two nested alpine/sub-alpine basins. More specifically, the aim is to link hydrograph interpretations to results of hydrochemistry during rain storms in order to understand better short term hydrochemical fluxes and variability in solute sources. Hydrograph separation was undertaken by using four hydrological tracers; electrical conductivity, concentration of silica, and the stable environmental isotopes oxygen-18 and deuterium. The different methods predicted consistent high pre-storm water contribution for the lower station at peak flow (60%-90%) but less consistent results were found at the upper basin outlet (25%-90%). The chemical characteristics of the stream water have been analyzed using three different approaches, namely; statistical, mass balance, and thermodynamic. Linear correlation was used to investigate the statistical association between discharge and the individual chemical species. The mass balance approach was used to correlate stoichiometry of the bedrock mineralogy to dissolved constituents in the stream water. Finally, a thermodynamic technique was used to evaluate to what extent the stream water could be represented as an equilibrium system and how this changed over the course of the storm. The results from these methods showed that the stream water variability was caused almost entirely by dilution from rain water input. / Arts, Faculty of / Geography, Department of / Graduate

>Magister Scientiae - MSc / Endocrine disrupting compounds (EDCs) are exogenous compounds/chemicals in the environment that interfere with the synthesis, secretion, distribution and function or elimination of natural hormones in the body. Environmental estrogens are a subclass of EDCs that may mimic or inhibit the effect of endogenous estrogen and can therefore influence developmental and reproductive health in humans and animals. EDCs have been reported to adversely affect the reproductive, immune, endocrine and nervous systems of wildlife and humans. The effects of EDCs include gonadal abnormalities, altered male/female sex ratios, reduced fertility and cancers of the male and female reproductive tract to mention a few. These effects are difficult to detect. Although it is essential to screen for EDCs in aqueous environmental samples, most countries have failed to implement this as part of their routine water quality monitoring programs due to various constraints such as the high cost of assays and the lack of infrastructure and skills required to do the assays. Therefore, there is a clear need for more user-friendly, more economically viable and time saving assays that can be used for routine monitoring of environmental EDCs. The aim of this study was to investigate the comparison between chemical and tissue culture methods to monitor environmental estrogens. 28 environmental water samples were collected from various sites around South Africa and analyzed for EDCs using a battery of rapid in vitro tests. Samples collected for the current study were selected based on various human impacts and also to give approximately 50% high and 50% low estrogen values. The 28 environmental water samples were separated into two groups based on the estradiol ELISA. The estradiol ELISA was chosen because estradiol is the principal estrogen found in all mammalian species during their reproductive years. For this separation, an estradiol level of 5 pg/ml was used as cut-off. Of the 28 samples investigated, 15 had estradiol levels higher than 5 pg/ml and were designated as high estradiol. The remaining 13 samples contained estradiol at 5 pg/ml or less and they were designated as low estradiol. The first objective of this study was to compare different rapid ELISAs for EDC monitoring to determine if the data obtained with these assays are similar/identical. The data obtained from the estrogenic ELISAs was related/similar and showed good correlation with each other. This is because the different estrogens are very similar and also due to the fact that the same sub-group in the population (the reproductively active females) is secreting these hormones. Therefore, an estradiol rapid assay was proposed as a first screening system for estrogens in samples. Even though there was a positive correlation between the estradiol rapid assay and testosterone rapid assay, separation of samples based on estradiol levels wasn’t a good predictor of testosterone levels in the samples. A testosterone rapid assay was therefore recommended as necessary to screen for androgens in samples. The positive correlation between the estradiol rapid assay and progesterone rapid assay was expected because both estradiol and progesterone are secreted and excreted by the same population sub-group (reproductively active females). This study also demonstrated a good predictability of separating samples containing progesterone using the estradiol ELISA. Progesterone is secreted by pregnant women, a sub-group of the reproductively active females. It is advised that a progesterone rapid assay be included to screen samples for progestogens. The second objective of this study was to compare estradiol rapid ELISAs with a bioassay for anti-androgenicity using mouse testicular cell cultures. The mouse testicular cell testosterone synthesis bioassay to monitor anti-androgenicity of the samples showed no correlation between the ELISA data for estrogens. This study shows that anti-androgenic effects need to be monitored independently because the data for estrogenic compounds cannot be used as a predictor for anti-androgenic effects. This demonstrated the need for the inclusion of a mouse testicular cell testosterone synthesis bioassay to screen for androgenicity and anti-androgenicity of water samples. In summary, due to the different mechanisms of action of EDCs, this study recommended a battery of assays to monitor for EDCs. The battery of assays suggested is: ●Estradiol ELISA as a rapid assay to screen for estrogens. ●Testosterone ELISA as a rapid assay to screen for androgens. ●Progesterone ELISA as a rapid assay to screen for progestogens. ●Mouse testicular cell testosterone synthesis bioassay to screen for androgenicity and anti-androgenicity.

O objetivo do presente estudo foi avaliar a viabilidade de cistos de Toxoplasma gondii em carnes suínas processadas por maturação úmida por 14, 21 e 28 dias a 0º (±1), assim como avaliar a distribuição dos cistos em órgãos e cortes comerciais de suínos experimentalmente infectados. Para tanto, dois experimentos foram conduzidos e em ambos, os suínos foram infectados com 3.000 oocistos do isolado TgCkBr57 (BrII). O lombo foi o corte muscular escolhido para sofrer o processo de maturação por ser o corte convencionalmente utilizado para o processo. O lombo direito de cada suíno foi submetido ao processo de maturação e o esquerdo foi mantido como controle, sem processamento. No Experimento 1 três suínos foram infectados. Dos lombos processados por maturação (n=3) e controle (n=3) realizou-se bioensaio em gatos e em camundongos e o período de maturação foi de 14 dias. Nesse ensaio também avaliou-se a distribuição de cistos teciduais pelo bioensaio em camundongos do cérebro, retina, língua, diafragma e coração e de cortes musculares: lombo (m. longissimus), copa (m. longissimus, spinalis dorsi, rhomboideus), filé mignon (m. psoas major), coxão-duro (m. biceps femoris), coxão mole (m. semimembranosus) e alcatra (m. gluteos medius). No Experimento 2 seis suínos foram infectados e foi realizado o bioensaio em camundongos dos lombos que ficaram sob maturação por 14 (n=2), 21 (n=2) e 28 (n=2) dias. Em ambos os experimentos, cistos de T. gondii presentes nos lombos permaneceram viáveis após 14 dias de maturação úmida, com confirmação pelo bioensaio em gatos e em camundongos. Nos períodos de 21 e 28 dias, pelo bioensaio observou-se que os camundongos não se infectaram, indicando que o processo inviabilizou os cistos. Quanto à distribuição dos cistos, estes foram isolados da copa, coração, diafragma e língua dos três suínos; do filé mignon, coxão duro e cérebro de dois suínos e da alcatra e lombo de um suíno. Nenhum camundongo infectou-se no bioensaio com o coxão mole e com a retina. Os resultados demonstram que a maturação em embalagem a vácuo por 14 dias em temperatura controlada (0ºC) não foi eficaz para inativação dos cistos de T. gondii, porém, o processo se mostrou eficiente quando a maturação foi feita por período igual ou superior a 21 dias. Cistos de T. gondii foram encontrados em praticamente todos os órgãos e cortes avaliados, demonstrando a ampla distribuição do parasita em suínos e a importância dessa espécie como fonte de infecção para o homem. / The objective of the present study was to evaluate the viability of Toxoplasma gondii cysts in pork processed by dry-aged for 14, 21 and 28 days at 0º (± 1), as well as to evaluate the distribution of T. gondii cysts in organs and commercial cuts of experimentally infected pigs. For that, two experiments were conducted ans in both the pigs were infected with 3.000 oocysts of the isolate TgCkBr57 (BrII). The loin was the muscle chosen to undergo the dry-aged because it is the cut conventionally used for the process. The right loin of each pig was submitted to the dry-aged and the left was kept as control, without processing. In Experiment 1, three pigs were infected. From the loins processed by dry-aged (n=3) e controle (n=3), the bioassay was performed on cats and mice and the aged period of the loins was 14 days. This study, also avaluated the distribution of T. gondii tissue cysts by bioassay in mice of brain, retina, tongue, diaphragm and hear and of the muscles: loin (m. longissimus), coppa (m. longissimus, spinalis dorsi, rhomboideus), tender loin (m. psoas major), outside flat (m. biceps femoris), topside (m. semimembranosus) and top sirloin (m. gluteos medius) was evaluated. In Experiment 2, six pigs were infected and the bioassay was performed in mice of the loins aged for 14 (n=2), 21 (n=2) and 28 (n=2) days. In both experiments, T. gondii cysts of the loins remained viable after 14 days of dry-aged, confirmed by bioassay in cats and mice. At 21 and 28 days, the bioassay showed that the mice did not become infected, indicating that the process made the cysts unfeasible. As to the distribution of the cysts, T. gondii was isolated from the coppa, heart, diaphragm and tongue of the three pigs, to the tender loin, outside flat and brain of two pigs and to the top sirloin and loin of one pig. No mice were infected in the bioassay with the topside and with the retina. The results demonstrate that the dry-aged for 14 days at controlled temperature (0ºC) was not effective for inactivation of T. gondii cysts, however, the process was efficient when the dry-aged was done for a period egual or righter than 21 days. Toxoplasma gondii cysts were found in practically all organs and cuts evaluated, demonstrating the wide distribution of the parasite in pigs and the importance of this species as a source of infection for man.

Vor dem Hintergrund der zunehmenden Prävalenz von Adipositas ist die Erforschung der zugrunde liegenden Pathomechanismen, unter anderem der Leptinresistenz, von großer Bedeutung. Der Schwerpunkt der vorliegenden Arbeit lag in der Etablierung und Validierung einer Methode zur Bestimmung der Bioaktivität von Leptin in Serum. Aufgrund des Vorhandenseins von Leptinbindungsproteinen erscheint nur der Teil des Serumleptins funktionell entscheiden, der tatsächlich den Leptinrezeptor erreicht und eine Bioantwort im Sinne einer Gewichtsreduktion auszulösen vermag. Der Bioassay basiert auf der Nutzung von Luciferase als Reportergen im HEK-293-Zellkulturmodell. Während der Validierung konnte gezeigt werden, dass der Test ein sensitives und spezifisches Verfahren zur Messung von Leptin im Serum darstellt. Problematisch blieb die Reproduzierbarkeit der Messergebnisse, so dass deren Interpretation im Vergleich mit Ergebnissen anderer Verfahren zum Nachweis von Leptin (Radioimmunoassay) sinnvoll erscheint. Der Luciferase-Bioassay wurde zur Bestimmung des Leptins im Serum adipöser Kinder eingesetzt, wobei eine statistisch signifikante Korrelation zwischen deren Bioaktivität und klinischen Markern der Adipositas gefunden wurde.

O objetivo do presente estudo foi avaliar a viabilidade de cistos de Toxoplasma gondii em carnes suínas processadas por maturação úmida por 14, 21 e 28 dias a 0º (±1), assim como avaliar a distribuição dos cistos em órgãos e cortes comerciais de suínos experimentalmente infectados. Para tanto, dois experimentos foram conduzidos e em ambos, os suínos foram infectados com 3.000 oocistos do isolado TgCkBr57 (BrII). O lombo foi o corte muscular escolhido para sofrer o processo de maturação por ser o corte convencionalmente utilizado para o processo. O lombo direito de cada suíno foi submetido ao processo de maturação e o esquerdo foi mantido como controle, sem processamento. No Experimento 1 três suínos foram infectados. Dos lombos processados por maturação (n=3) e controle (n=3) realizou-se bioensaio em gatos e em camundongos e o período de maturação foi de 14 dias. Nesse ensaio também avaliou-se a distribuição de cistos teciduais pelo bioensaio em camundongos do cérebro, retina, língua, diafragma e coração e de cortes musculares: lombo (m. longissimus), copa (m. longissimus, spinalis dorsi, rhomboideus), filé mignon (m. psoas major), coxão-duro (m. biceps femoris), coxão mole (m. semimembranosus) e alcatra (m. gluteos medius). No Experimento 2 seis suínos foram infectados e foi realizado o bioensaio em camundongos dos lombos que ficaram sob maturação por 14 (n=2), 21 (n=2) e 28 (n=2) dias. Em ambos os experimentos, cistos de T. gondii presentes nos lombos permaneceram viáveis após 14 dias de maturação úmida, com confirmação pelo bioensaio em gatos e em camundongos. Nos períodos de 21 e 28 dias, pelo bioensaio observou-se que os camundongos não se infectaram, indicando que o processo inviabilizou os cistos. Quanto à distribuição dos cistos, estes foram isolados da copa, coração, diafragma e língua dos três suínos; do filé mignon, coxão duro e cérebro de dois suínos e da alcatra e lombo de um suíno. Nenhum camundongo infectou-se no bioensaio com o coxão mole e com a retina. Os resultados demonstram que a maturação em embalagem a vácuo por 14 dias em temperatura controlada (0ºC) não foi eficaz para inativação dos cistos de T. gondii, porém, o processo se mostrou eficiente quando a maturação foi feita por período igual ou superior a 21 dias. Cistos de T. gondii foram encontrados em praticamente todos os órgãos e cortes avaliados, demonstrando a ampla distribuição do parasita em suínos e a importância dessa espécie como fonte de infecção para o homem. / The objective of the present study was to evaluate the viability of Toxoplasma gondii cysts in pork processed by dry-aged for 14, 21 and 28 days at 0º (± 1), as well as to evaluate the distribution of T. gondii cysts in organs and commercial cuts of experimentally infected pigs. For that, two experiments were conducted ans in both the pigs were infected with 3.000 oocysts of the isolate TgCkBr57 (BrII). The loin was the muscle chosen to undergo the dry-aged because it is the cut conventionally used for the process. The right loin of each pig was submitted to the dry-aged and the left was kept as control, without processing. In Experiment 1, three pigs were infected. From the loins processed by dry-aged (n=3) e controle (n=3), the bioassay was performed on cats and mice and the aged period of the loins was 14 days. This study, also avaluated the distribution of T. gondii tissue cysts by bioassay in mice of brain, retina, tongue, diaphragm and hear and of the muscles: loin (m. longissimus), coppa (m. longissimus, spinalis dorsi, rhomboideus), tender loin (m. psoas major), outside flat (m. biceps femoris), topside (m. semimembranosus) and top sirloin (m. gluteos medius) was evaluated. In Experiment 2, six pigs were infected and the bioassay was performed in mice of the loins aged for 14 (n=2), 21 (n=2) and 28 (n=2) days. In both experiments, T. gondii cysts of the loins remained viable after 14 days of dry-aged, confirmed by bioassay in cats and mice. At 21 and 28 days, the bioassay showed that the mice did not become infected, indicating that the process made the cysts unfeasible. As to the distribution of the cysts, T. gondii was isolated from the coppa, heart, diaphragm and tongue of the three pigs, to the tender loin, outside flat and brain of two pigs and to the top sirloin and loin of one pig. No mice were infected in the bioassay with the topside and with the retina. The results demonstrate that the dry-aged for 14 days at controlled temperature (0ºC) was not effective for inactivation of T. gondii cysts, however, the process was efficient when the dry-aged was done for a period egual or righter than 21 days. Toxoplasma gondii cysts were found in practically all organs and cuts evaluated, demonstrating the wide distribution of the parasite in pigs and the importance of this species as a source of infection for man.