A avidez de anticorpos específicos anti-toxoplasma da classe IgG e sua utilização na diferenciação entre toxoplasmose recente e crônica em caprinos

Foi avaliada a avidez de anticorpos IgG como marcador sorológico de infecção recente e crônica pelo Toxoplasma gondii, através da dissociação do complexo antígeno-anticorpo com uréia. A avaliação foi realizada medindo-se a percentagem de queda (%Q) da absorbância, pelo ELISA, após a lavagem do complexo Ag - Ac formado com solução de uréia. Ficou determinado que a concentração dc uréia 9 Molar (M) foi a que melhor diferenciou infecções recentes e crônicas de cabras experimentalmente infectadas com Toxoplasma gondii, e que a %Q da absorbância decresce com o tempo de infecção do animal, tornando-se estável em torno do 100º. dia após a inoculação. Em um grupo de 116 amostras de soro, coletadas de caprinos naturalmente infectados, foram demonstrados perfis característicos de toxoplasmose recente, crônica ou em fase de transição. Os perfis foram previamente determinados pela avaliação da evolução dos níveis de anticorpos IgG pela reação de imunofluorescência indireta. Os animais caracterizados como portadores de toxoplasmose crônica apresentaram uma %Q da absorbância de 26,32 ± 10,84. Para toxoplasmose recente a %Q da absorbância observada foi de 77,61 ± 13,89 e para o perfil de transição, 46,22 ± 11,94

Toxoplasma - specific IgG antibodies avidity ELISA and its role in the differentiation between recent and chronic toxoplasmosis in goats

For serological characterization of recent Toxoplasma infection in goats low avidity IgG specific antibodies were studied. Avidity was evaluated as the decrease of absorbance in ELISA test after treating plates with 3, 6 and 9M urea, as a dissociating solution of low avidity antigen-antibody complexes. A concentration of 9M urea best discriminated between recent and old infections in goats experimentally infected with Toxoplasma gondii. Absorbance after treating plates with 9M urea decreased in relation to the duration of infection, levelling off about the 100th day after inoculation. In a group of 116 serum samples from naturally infected goats patterns of recent, transitional or chronic toxoplasmosis were demonstrated. Serological patterns were previously determined by immunofluorescent assay. For chronic toxoplasmosis infection, the observed decrease of absorbance was 26.32 ± 10.84. For recent toxoplasmosis it was 77.61 ± 13.89, and for transitional pattern, 46.22 ± 11.94.Foi avaliada a avidez de anticorpos IgG como marcador sorológico de infecção recente e crônica pelo Toxoplasma gondii, através da dissociação do complexo antígeno-anticorpo com uréia. A avaliação foi realizada medindo-se a percentagem de queda (%Q) da absorbância, pelo ELISA, após a lavagem do complexo Ag - Ac formado com solução de uréia. Ficou determinado que a concentração dc uréia 9 Molar (M) foi a que melhor diferenciou infecções recentes e crônicas de cabras experimentalmente infectadas com Toxoplasma gondii, e que a %Q da absorbância decresce com o tempo de infecção do animal, tornando-se estável em torno do 100º. dia após a inoculação. Em um grupo de 116 amostras de soro, coletadas de caprinos naturalmente infectados, foram demonstrados perfis característicos de toxoplasmose recente, crônica ou em fase de transição. Os perfis foram previamente determinados pela avaliação da evolução dos níveis de anticorpos IgG pela reação de imunofluorescência indireta. Os animais caracterizados como portadores de toxoplasmose crônica apresentaram uma %Q da absorbância de 26,32 ± 10,84. Para toxoplasmose recente a %Q da absorbância observada foi de 77,61 ± 13,89 e para o perfil de transição, 46,22 ± 11,94

Use of dried blood sample for serological diagnosis of toxoplasmosis in goats

Eluates of 183 dried caprine blood samples on filter paper were tested for anti-Toxoplasma gondii antibodies, using IFA, ELISA and dot-ELISA, and compared with results obtained by direct observations on comparable sera, usin the same tests. The results has showed that papers with dried blood can be preserved for up to 45 days at room temperature and for six months at 4°C, provided they are protected against humid conditions by using desiccating agents such as silica-gel. Comparison between serum and eluate reactions revelated 97-100% correspondance. The antibody titers remained constant throughout the study period.Foi realizada pesquisa de anticorpos anti-Toxoplasma gondii em 183 amostras de sangue dessecado em papel de filtro utilizando as reações de imunofluorescência indireta ELISA e dot-ELISA, tomando como referência os resultados obtidos nos soros. A análise dos resultados demonstrou que papéis com sangue dessecado podem ser conservados por um período mínimo de 45 dias à temperatura ambiente e por seis meses a 4°C, desde que mantidos livres de umidade pela utilização de agentes dessecantes como a sílica-gel. A reprodutibilidade das reações, avaliada por meio da curva dos títulos de anticorpos no decorrer do tempo após a coleta do sangue em papéis de filtro, demonstrou uma concordância de 97 a 100% entre os resultados obtidos nos soros e eluatos. Os títulos de anticorpos permaneceram estáveis durante o período observado. Os resultados obtidos com eluato de sangue dessecado foram semelhantes na RIFI, ELISA e dot-ELISA, indicando que qualquer uma das três reações pode ser utilizada em eluatos de sangue dessecado para o diagnóstico da toxoplasmose caprina

Detección molecular de Toxoplasma gondii en carnes para consumo humano en Ibagué, Colombia

Introduction: Toxoplasma gondii is a parasite with great zoonotic potential. It can infect a broad range of warm-blooded hosts (including livestock) and causes significant losses in the industry. In humans, it has been described as a pathogen in immunosuppressed people, it affects the fetus development in congenital infections, and is associated with various behavioral disorders in healthy people. Humans can acquire T. gondii by consuming undercooked, contaminated meat.Objective: To determine T. gondii positivity (currently unknown) in meat for human consumption (i.e., beef, chicken, and pork) in the city of Ibague, Colombia.Materials and methods: We used conventional nested PCR and the T. gondii B1 gene sequence as amplification target. We collected samples of meat (N=186) sold in the urban area of Ibagué (62 beef, 62 chicken, and 62 pork samples) and determined the T. gondii positivity percentage for each type of meat.Results: The study found an average of 18.8% positivity for all meat samples, pork having the highest percentage (22.5%; 14/62), followed by beef (19.3%; 12/62) and chicken (14.5%; 9/62). The best-amplified products were sequenced by macrogen and aligned with the B1 gene sequences in GenBank, thereby confirming their identity.Conclusions: This is the first study of T. gondii prevalence in meat for human consumption carried out in the city of Ibagué and the department of Tolima. All three types of meat sampled represent a risk for local human infectionIntroducción. Toxoplasma gondii es un parásito con gran potencial zoonótico que puede infectar un amplio rango de huéspedes de sangre caliente, incluidos los animales del sector pecuario, lo que causa pérdidas a la industria. En el humano, es patógeno en personas inmunosuprimidas y afecta el desarrollo del feto en infecciones congénitas. Además, se asocia con diversos trastornos del comportamiento en personas sanas. El humano puede adquirir T. gondii al consumir carnes contaminadas mal cocidas.Objetivo. Determinar la positividad de T. gondii en carnes de consumo humano (res, pollo y cerdo) en Ibagué, Colombia.Materiales y métodos. Se utilizó la PCR convencional anidada y la secuencia del gen B1 de T. gondii como blanco de amplificación. Se tomaron 186 muestras de carne comercializada en la zona urbana de Ibagué (62 de res, 62 de pollo y 62 de cerdo) y se obtuvo el porcentaje de positividad en cada tipo de carne evaluada.Resultados. Se encontró un porcentaje de positividad de 18,8 % en las muestras, siendo la carne de cerdo la del mayor porcentaje (22,5 %; 14/62), seguida por las muestras de carne de res (19,3 %; 12/62) y de pollo (14,5 %; 9/62). Los mejores productos amplificados fueron secuenciados en Macrogen, y alineados con las secuencias del gen B1 depositadas en el GenBank, con lo que se confirmó su identidad.Conclusiones. Este es el primer estudio sobre prevalencia de T. gondii en carnes para consumo humano en Ibagué y el departamento del Tolima. Se demostró que los tres tipos de carne representan un riesgo para la infección en humanos a nivel local

Soroprevalência de Brucella ovis, Brucella lisa, leptospirose, toxoplasmose e Maedi-Visna em ovinos abatidos em Minas Gerais, Brasil

The present study aimed to estimate the prevalence of antibodies against Brucella ovis-epididymitis, smooth-Brucella, leptospirosis, toxoplasmosis and Maedi-visna in sheep slaughtered in Minas Gerais, Brazil and to study their simultaneous occurrence, including caseous lymphadenitis, at sheep and flock levels. The study was conducted at a sheep slaughterhouse with Federal Inspection Service. Sera from 594 animals from 21 flocks were collected, in 2007. The agar gel immunodiffusion (AGID) was employed to detect anti-B. ovis and anti-Maedi Visna antibodies, whereas Rose Bengal (RB) and the 2-mercaptoethanol test (2ME) were used to test anti-smooth Brucella antibodies. For the detection of anti-Leptospira antibodies, sera were examined by microscopic agglutination test (MAT), while for the detection of IgG antibodies to Toxoplasma gondii ELISA was used. Prevalence of antibodies against smooth Brucella, B. ovis-epididimitis, Leptospira spp., toxoplasmosis and Maedi-Visna found in sheep from Minas Gerais was 0.00%, 24.04%, 25.96%, 10.46% and 3.08%, respectively; whereas the seroprevalence in flocks was 0.00%, 80.95%, 90.48%, 71.43% and 23.81%, respectively. Moreover, when data on antibodies anti-Corynebacterium pseudotuberculosis, previously obtained, were included, about 60% of the flocks showed animals that were exposed to four or more of the studied agents. However, only 25.47% of the sheep exhibited simultaneously antibodies against more than one pathogen. Thus, data from the present study on sheep slaughtered in Minas Gerais, Brazil, showed no antibodies to smooth-Brucella and a low frequency of antibodies anti-Maedi Visna lentivirus, and a high and widespread seroprevalence of B. ovis, Leptospira spp., and T. gondii among animals and flocks.O presente estudo teve como objetivo estimar a prevalência de anticorpos contra Brucella ovis (epididimite ovina), Brucella lisa, leptospirose, toxoplasmose e Maedi-visna em ovinos abatidos em Minas Gerais, Brasil, e estudar sua ocorrência simultânea, incluindo linfadenite caseosa, nos ovinos e nos rebanhos. O estudo foi realizado em um abatedouro de ovinos com Serviço de Inspeção Federal. Soros de 594 animais de 21 rebanhos foram coletados, em 2007. A imunodifusão em gel de ágar (IDGA) foi empregada para detectar anticorpos anti-B. ovis e anticorpos anti-Maedi Visna, enquanto o teste do antígeno acidificado tamponado (AAT) e o teste de 2-mercaptoetanol (2ME) foram utilizados para testar anticorpos anti-Brucella lisa. Para a detecção de anticorpos anti-Leptospira, os soros foram examinados pelo teste de aglutinação microscópica (MAT), enquanto que para a detecção de anticorpos IgG para Toxoplasma gondii, foi usado o ELISA. A prevalência de anticorpos anti-Brucella lisa, B. ovis, Leptospira spp., toxoplasmose e Maedi-Visna encontrados em ovinos de Minas Gerais foi de 0,00%, 24,04%, 25,96%, 10,46% e 3,08%, respectivamente; enquanto a soroprevalência em rebanhos foi de 0,00%, 80,95%, 90,48%, 71,43% e 23,81%, respectivamente. Além disso, quando dados de anticorpos anti-Corynebacterium pseudotuberculosis, previamente obtidos, foram incluídos, cerca de 60% dos rebanhos apresentaram animais expostos a quatro ou mais dos agentes estudados. No entanto, apenas 25,47% dos ovinos exibiram simultaneamente anticorpos contra mais de um patógeno. Assim, os dados do presente estudo sobre ovinos abatidos em Minas Gerais, Brasil, mostram que ausência de anticorpos anti-Brucella lisa e baixa frequência de anticorpos anti-Maedi Visna, e uma soroprevalência alta e generalizada de B. ovis, Leptospira spp. e T. gondii entre animais e rebanhos

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt