Detección y cuantificación de SARS-CoV-2 en plantas de tratamiento de aguas residuales de diferentes ciudades de Chile: hacia la implementación de una vigilancia centinela permanente

Introducción: La cuantificación de SARS-CoV-2 en aguas residuales es una herramienta que permite determinar la tendencia de la circulación viral en un área geográfica determinada. Objetivo: Cuantificar el virus SARS-CoV-2 en 15 plantas de tratamiento de aguas residuales en diferentes ciudades de Chile para establecer una comparación con las variables de: i) casos activos por cada 100.000 habs.; ii) positividad diaria (casos nuevos); y iii) fases del plan de confinamiento. Metodología: SARS-CoV-2 se concentró a partir de muestras de aguas residuales. Para obtener el número de genomas del virus por litro se realizó una cuantificación absoluta utilizando qRT-PCR. Resultados: Entre enero y junio de 2021 se procesaron 253 muestras, siendo todas positivas para la presencia del virus. Asimismo, se logró determinar que la tasa de casos activos por cada 100.000 habs. es la variable que mejor se ajusta a las tendencias obtenidas con la cuantificación de la carga viral en las aguas residuales. Conclusiones: La cuantificación de SARS-CoV-2 en las aguas residuales de manera permanente es una herramienta eficiente para determinar la tendencia del virus en un área geográfica determinada y, en conjunto con una vigilancia genómica, puede constituirse en una vigilancia centinela ideal generando alertas sobre futuros brotes.

Background: The quantification of SARS-CoV-2 in wastewater is a tool that allows determining the trend of viral circulation in a particular geographical area. Aim: To quantify the SARS-CoV-2 virus in 15 wastewater treatment plants in different Chilean cities to establish a comparison with the variables of: i) Active cases per 100,000 inhabitants; ii) daily positivity (novel cases); and iii) phases of the lockdown strategy. Methods: SARS-CoV-2 was concentrated from wastewater samples. To obtain the number of virus genomes per liter, absolute quantification was performed using qRT-PCR. Results: Between January and June 2021, 253 samples were processed, all of which were positive for the presence of the virus. Likewise, it will be determined that the rate of active cases per 100,000 inhabitants is the variable that best fits the trends obtained with the quantification of the viral load in wastewater. Conclusions: The quantification of SARS-CoV-2 in wastewater as a continuous strategy is an efficient tool to determine the trend of the viral circulation in a delimited geographical area and, combined with genomic surveillance, it can constitute an ideal sentinel surveillance alert on future outbreaks.

Characterization of the milk production chain and study of the dairy herd health in the city of Codó, Maranhão / Caracterização da cadeia produtiva do leite e estudo da sanidade do rebanho leiteiro no município de Codó, Maranhão

In order to characterize the milk production chain and study the dairy herd health in the city of Codó, state of Maranhão, Brazil, a checklist was applied and tests were carried out to detect clinical and subclinical mastitis using mastitis test strip cup and the California mastitis test (CMT), from June to August 2019, in 295 dairy cows from 20 farms. Water and milk samples were collected for microbiological analysis. It was observed that herdsmen do not have adequate knowledge about good agricultural practices. As for milking, only 60% are performed in corrals with coverage, and as for the floor, 60% are made of clay and 40% are made of concrete. In 35% of the properties, the water used in milking management comes from wells and the rest from dams. Pre and postdipping practices, CMT, mastitis strip cup test and the adoption of a mastitis control program are not carried out on any of the properties Two cows tested positive for subclinical mastitis and one cow tested positive for tuberculosis. In the microbiological analysis of the milk, a high count of total coliforms and thermotolerants was obtained, with values between 23 to > 1,100 MPN/mL and < 3.0 to > 1,100 MPN/mL, respectively. The presence of coagulase positive staphylococci was also observed in 25% of the samples. The water samples also showed high contamination by total coliforms between 4.1 to > 2.419.6 MPN/mL and 40% showed the presence of Escherichia coli. These results reflect the need for more investments in technical assistance and technical training for these producers.(AU)

Este estudo teve como objetivo caracterizar a cadeia produtiva do leite e estudar a sanidade do rebanho leiteiro do município de Codó, estado do Maranhão, Brasil, através da aplicação de um checklist e da realização de exames para detecção de mastite clínica e subclínica pelos métodos da caneca do fundo escuro e California mastitis test (CMT), no período de junho a agosto de 2019, em 295 vacas leiteiras procedentes de 20 propriedades. Coletaram-se amostras de leite e água da ordenha para análises microbiológicas. Observou-se que os ordenhadores não possuem conhecimento adequado sobre as boas práticas agropecuárias. Quanto à realização da ordenha, apenas 60% realizam-na em currais com cobertura; quanto ao piso, 60% são de terra batida e 40% de concreto. A água utilizada no manejo da ordenha é proveniente de poços em 35% das propriedades e as demais de açudes. As práticas de pré e pós-dipping e os testes CMT e caneca de fundo escuro e a adoção de programa de controle da mastite não são realizados em nenhuma das propriedades. Diagnosticaram-se duas vacas com mastite subclínica e uma com tuberculose e verificou-se elevada contaminação por coliformes totais e termotolerantes nas análises microbiológicas do leite, variando entre 23 NMP/mL a >1.100 NMP/mL e < 3.0 a > 1.100 NMP/mL, respectivamente, e presença de estafilococos coagulase positivos em 25% das amostras. As amostras de água também apresentaram elevada contaminação por coliformes totais entre 4,1 NMP/mL a > 2.419,6 NMP/mL e 40% apresentaram presença de Escherichia coli. Esses resultados refletem a necessidade de mais investimentos em assistência técnica e treinamento técnico para esses produtores.(AU)

Interventions to Reduce Stigma Related to People who Use Drugs: Systematic Review / Intervenções para Redução do Estigma Relacionado às Pessoas que Usam Drogas: Revisão Sistemática / Intervenciones para la Reducción del Estigma Relacionado a las Personas que Usan Drogas: Revisión Sistemática

Abstract Interventions to reduce stigma related to people who use drugs can improve their quality of life and adherence to treatment. This review aimed to identify and analyze studies on interventions to reduce the stigma related to people who use drugs. Searches were performed in the following databases, without period delimitation: PubmMed, APA PsycNET, ScienceDirect, Web of Science, and VHL. Considering secondary referencing, we identified a total of 5,488 records. This review covered 28 articles and was based on PRISMA. Although target audiences comprising students and health professionals were predominant, as well as the use of quantitative methods, sample size and interventions varied among studies. The selected studies present considerable methodological limitations. We concluded that no evidence confirms the effectiveness of the proposed interventions and which of them should continue to be applied for this specific aim. It is essential to invest in approaches other than those traditionally adopted.

Resumo Intervenções para a redução do estigma relacionado às pessoas que usam drogas podem melhorar sua qualidade de vida e adesão ao tratamento. Esta revisão teve por objetivo identificar e analisar estudos de intervenções para reduzir o estigma relacionado às pessoas que usam drogas. Foram realizadas buscas nas bases PubMed, APA PsycNET, ScienceDirect, Web of Science e BVS, sem delimitação de ano. Foram identificados 5.488 registros somados à busca secundária de referências. Esta revisão abrange 28 artigos e foi baseada no PRISMA. Houve variação no tamanho da amostra e intervenções, embora o público-alvo de estudantes e profissionais da saúde e o uso de métodos quantitativos tenham sido predominantes. Limitações metodológicas consideráveis foram identificadas nos estudos. Portanto, conclui-se que não há evidências que permitam inferir que as intervenções propostas são efetivas e quais delas deveriam continuar a ser empregadas para essa finalidade. É imprescindível investir em abordagens distintas das adotadas tradicionalmente.

Resumen Las intervenciones para la reducción del estigma relacionado a las personas que usan drogas pueden mejorar su calidad de vida y la adherencia al tratamiento. Esta revisión tuvo como objetivo identificar y analizar los estudios sobre las intervenciones para la reducción del estigma relacionado a las personas que usan drogas. Se realizaron búsquedas en las bases de datos PubMed, APA PsycNET, ScienceDirect, Web of Science y BVS, sin delimitación del año. Se identificaron 5.488 registros sumados a la búsqueda secundaria de referencias. Esta revisión abarca 28 artículos y se basó en PRISMA. Hubo una variación en el tamaño de la muestra y las intervenciones, aunque fueron predominantes el público objetivo de estudiantes y profesionales de la salud y el uso de métodos cuantitativos. Se identificaron limitaciones metodológicas considerables en los estudios. Se concluye que no hay evidencias de que las intervenciones propuestas son efectivas y cuáles deberán seguir siendo utilizadas para este propósito. Es esencial invertir en enfoques distintos de los adoptados tradicionalmente.

Trifolium pratense: Friable calli, cell culture protocol and isoflavones content in wild plants, in vitro and cell cultures analyzed by UPLC

Abstract Trifolium pratense L., Fabaceae, is a rich source of isoflavones and has become the focus of several studies related to its phytoestrogenic activity. The aim of this study was to establish germination and cell cultures protocol for T. pratense and quantify isoflavones content in cell cultures, in vitro cultured and wild plants harvested in two different seasons. Murashige Skoog medium supplemented with naphthalene acetic acid and kinetin was able to produce the highest formation of friable calli. Calli cultures were analyzed qualitatively after 60 days of culture, and in vitro plants after 30, 45 and 60 days of cultivation. The chemical analysis was performed by ultra performance liquid chromatography, using the linearity curves of daidzein, genistein, formononetin and biochanin A as standards. The concentrations of isoflavones detected in wild plants were different in the two harvest periods and contrasted in content when compared to the in vitro plants. Cell cultures exhibited diverse profiles and concentration of isoflavones, none of which presented the isoflavonoid biochanin A. Pectinase was used to promote reduction of clumps and ended up altering the characteristics of secondary metabolites production in some cultures. Formononetin showed higher concentration in wild red clover samples (15.407 mg g-1), and in the in vitro grown plants the highest concentration was daidzein (17.591 mg g-1) at 60 days. The methods used for this research were effective, and the red clover plants of the analyzed variety can be cultivated in vitro aiming the commercial productivity by having contents greater than or equal to the wild plants in the periods studied, even without the use of elicitors during the cultivation.

Evaluation of physicochemical properties as supporting information on quality control of raw materials and veterinary pharmaceutical formulations / 药物分析学报

This study aimed to show that the physicochemical proprieties obtained by Fourier transform infrared spectroscopy (FTIR), thermogravimetry (TG), and scanning electronic microscopy (SEM) can be useful tools for evaluating the quality of active pharmaceutical ingredients (APIs) and pharmaceutical products. In addition, a simple, sensitive, and efficient method employing HPLC-DAD was developed for simulta-neous determination of lidocaine (LID), ciprofloxacin (CFX) and enrofloxacin (EFX) in raw materials and in veterinary pharmaceutical formulations. Compounds were separated using a Gemini C18 (250 mm × 4.6 mm, 5 μm) Phenomenex ? column, at a temperature of 25 °C, with a mobile phase containing 10 mM of phosphoric acid (pH 3.29): acetonitrile (85.7:14.3, v/v) and a flow rate of 1.5 mL/min. Physicochemical characterization by TG, FTIR, and SEM of raw materials of LID, CFX, and EFX provided information useful for the evaluation, differentiation, and qualification of raw materials. Finally, the HPLC method was proved to be useful for evaluation of raw material and finished products, besides satisfying the need for an analytical method that allows simultaneous determination of EFX, CFX, and LID, which can also be extended to other matrices and applications.

Murine Experimental Root Canal Infection: Cytokine Expression in Response to F. nucleatum and E. faecalis

Abstract The aim of this study was to evaluate the gene expression of proinflammatory (RANKL, TNF-a and IFN-g) and regulatory (TGF-b and IL-10) cytokines as reaction to experimental infection by mono or bi-association of Fusobacterium nucleatum (ATCC 10953) and Enterococcus faecalis (ATCC 19433). F. nucleatum and E. faecalis, either in mono- or bi-association were inoculated into the root canal system (RCS) of Balb/c mice. Animals were sacrificed at 10 and 20 days after infection and periapical tissues surrounding the root were collected. The mRNA expression of the cytokines RANKL, TNF-a, IFN- g, TGF-b and IL-10 was assessed using real-time PCR. The Kruskal-Wallis test was used for statistical analysis. F. nucleatum mono-infection induced high expression of RANKL and TNF-a, while its modulation was due to IL-10. High expression of IFN-g at day 20 was up-regulated by E. faecalis and RANKL; TNF-a was up-regulated by an independent mechanism via IL-10 and TGF-b. Bi-association (F. nucleatum and E. faecalis) stimulated high expression of RANKL, TNF-a and IFN-g, which seemed to be modulated by TGF-b 20 days later. The gene expression of proinflammatory cytokines was more prominent in the earlier periods of the experimental periapical infection, which concomitantly decreased in the later period. This expression may be regulated by IL-10 and TGF-b in an infection-specific condition

Resumo O objetivo deste trabalho foi avaliar a expressão gênica de citocinas pró-inflamatórias (RANKL, TNF-a e IFN-g) e regulatórias (TGF-b e IL-10) em resposta à infecção experimental por Fusobacterium nucleatum (ATCC 10953) e Enterococcus faecalis (ATCC 19433) como mono-infecção ou em bi-associação. F. nucleatum e E. faecalis foram inoculados no sistema de canais radiculares de camundongos Balb/c, tanto isoladas como em bi-associação. Os animais foram sacrificados em 10 e 20 dias após a infecção, e os tecidos periapicais foram coletados. As expressões do mRNA das citocinas RANKL, TNF-a, IFN-g, TGF-b e IL-10 foram analisadas por meio do real-time PCR. O teste de Kruskal-Wallis foi utilizado para análise estatística. A mono-infecção com F. nucleatum induziu alta expressão de RANKL e TNF-a, enquanto sua modulação ocorreu devido à IL-10. A alta expressão de IFN-g no dia 20 foi regulada positivamente por E. faecalis e RANKL; TNF-a foi regulada positivamente por um mecanismo independente via IL-10 e TGF-b. A bi-associação (F. nucleatum e E. faecalis) estimulou uma alta expressão de RANKL, TNF-a e IFN-g, que parece ser modulada por TGF-b após 20 dias. A expressão gênica de citocinas pró-inflamatórias foi mais proeminente nos estágios iniciais da infecção periapical experimental, com concomitante redução no período tardio. Este fenômeno pode ser regulado por IL-10 e TGF-b em uma condição de infecção específica.