Use of laboratory methodologies to confirm an outbreak of acute diarrheal disease caused by multiple pathogens / Uso de metodologias laboratoriais para confirmação de surto de doença diarreica aguda causada por múltiplos patógenos

The aim of this study was to investigate an outbreak caused by protozoa, which occurred in a municipality in the Brazil southern region. The investigations were carried out analyzing 47 fresh stool samples and 26 water samples by parasitological and molecular methods, as well as, direct immunofluorescence. After the filtrations of water samples and purification of stool samples, the concentrates were evaluated microscopically for presence of parasites. Molecular analyses were performed by polymerase chain reaction (PCR) for DNA detection of Giardia spp., Cryptosporidium parvum, C. hominis and Cyclospora cayetanensis. Out of 26 water samples, 30.8% (8/26) had waterborne protozoa and C. cayetanensis was the most prevalent (15.5%). Out of the 47 stool samples, 23.4% (11/47) were infected with C. cayetanensis and Giardia spp. The results showed that backwash water samples from filters of the Water Treatment Station were contaminated with C. cayetanensis, C. hominis and Giardia spp., suggesting the contamination of water sources with human waste brought by sewage. These results show the importance of protozoa investigation in water and stool samples by laboratory methodologies principally in outbreaks causing acute diarrheal disease (AU).

O objetivo do presente estudo foi investigar um surto causado por protozoários, ocorrido em um município da região sul do Brasil. As investigações foram realizadas analisando 47 amostras de fezes frescas e 26 amostras de água por métodos parasitológicos, moleculares e de imunofluorscência direta. Após as filtrações das amostras de água e purificação das amostras de fezes, os concentrados foram avaliados microscopicamente a procura de parasitas. A seguir, foram analisadas, pela reação em cadeia da polimerase (PCR), a detecção de DNA de Giardia spp., Cryptosporidium parvum, C. hominis e Cyclospora cayetanensis. Das 26 amostras de água, 30,8% (8/26) apresentaram protozoários de veiculação hídrica, sendo que, C. cayetanensis foi o mais prevalente (15,5%). Das 47 amostras de fezes, 23,4% (11/47) estavam infectadas por C. cayetanensis e Giardia spp. Os resultados mostraram que as águas de retrolavagem dos filtros da Estação de Tratamento de Água estavam contaminadas com C. cayetanensis, C. hominis e Giardia spp. sugerindo a contaminação dos mananciais com dejetos humanos trazidos pelo esgoto. Estes resultados mostram a importância da investigação de protozoários em água e fezes por metodologias laboratoriais, principalmente em surtos que causam doença diarreica aguda (AU).

Genetic diversity of Trypanosoma cruzi strains isolated from chronic chagasic patients and non-human hosts in the state of São Paulo, Brazil

BACKGROUND Trypanosoma cruzi shows an exuberant genetic diversity. Currently, seven phylogenetic lineages, called discrete typing units (DTUs), are recognised: TcI-TcVI and Tcbat. Despite advances in studies on T. cruzi and its populations, there is no consensus regarding its heterogeneity. OBJECTIVES This study aimed to perform molecular characterisation of T. cruzi strains, isolated in the state of São Paulo, to identify the DTUs involved and evaluate their genetic diversity. METHODS T. cruzi strains were isolated from biological samples of chronic chagasic patients, marsupials and triatomines through culture techniques and subjected to molecular characterisation using the fluorescent fragment length barcoding (FFLB) technique. Subsequently, the results were correlated with complementary information to enable better discrimination between the identified DTUs. FINDINGS It was possible to identify TcI in two humans and two triatomines; TcII/VI in 19 humans, two marsupials and one triatomine; and TcIII in one human host, an individual that also presented a result for TcI, which indicated the possibility of a mixed infection. Regarding the strains characterised by the TcII/VI profile, the correlation with complementary information allowed to suggest that, in general, these parasite populations indeed correspond to the TcII genotype. MAIN CONCLUSIONS The TcII/VI profile, associated with domestic cycles and patients with chronic Chagas disease, was the most prevalent among the identified DTUs. Furthermore, the correlation of the study results with complementary information made it possible to suggest that TcII is the predominant lineage of this work.

Irrigation management in soybean crops influences the occurrence of nematodes in the soil / Manejo de irrigação na cultura da soja influencia a ocorrência de nematóides no solo

One of the main limitations of soybean production is related to water availability and organisms found in the soil. Under the hypothesis that soil moisture may influence the nematode population, this study aimed to verify the occurrence of nematodes associated with different irrigation management in soybean crops. The experiment was carried out in a randomized block design. Treatments consisted of a subplot scheme, with four replications. The plots consisted of ten irrigation managements divided into five irrigation frequencies (1,2,3,4, and 5 days) and five additional water depths (25, 50, 75, 100, and 125% of crop evapotranspiration ­ Etc). The subplots were composed of four different soybean cultivars (NA 5909 RR, AS 3680 IPRO, and Desafio RR ePower IPRO). The experiment analyzed the variables plant height, first pod insertion, number of plants, moisture, hundred-grain weight, yield, and occurrence of nematode eggs and adults. Nematodes influence all the production components analyzed, affecting mostly the hundred-grain weight, especially the nematode Helicotylenchus sp., although the Meloidogyne sp. population was larger. The phytonematode population reduces with the increase in soil moisture to levels close to the saturation, indirectly influencing the yield increase.

Uma das principais limitações da produção de soja está relacionada à disponibilidade de água e organismos encontrados no solo. Sob a hipótese de que a umidade do solo possa influenciar a população de nematoides, este estudo teve como objetivo verificar a ocorrência de nematóides associados a diferentes manejos de irrigação em lavouras de soja. O experimento foi conduzido em delineamento de blocos casualizados. Os tratamentos consistiram de um esquema de subparcelas, com quatro repetições. As parcelas consistiram de dez manejos de irrigação divididos em cinco freqüências de irrigação (1,2,3,4 e 5 dias) e cinco lâminas d'água (25, 50, 75, 100 e 125% da evapotranspiração da cultura - Etc). As subparcelas foram compostas por quatro diferentes cultivares de soja (NA 5909 RR, AS 3680 IPRO e Desafio RR ePower IPRO). O experimento analisou as variáveis altura da planta, inserção da primeira vagem, número de plantas, umidade, peso de cem grãos, produtividade e ocorrência de ovos de nematoides e adultos. Os nematóides influenciam todos os componentes de produção analisados, afetando principalmente o peso de cem grãos, especialmente o nematódeo Helicotylenchus sp., Embora o Meloidogyne sp. população era maior. A população de fitonematóides reduz com o aumento da umidade do solo para níveis próximos à saturação, influenciando indiretamente o aumento da produtividade.

Contribution of morphological traits for grain yield in common bean / Contribuição de caracteres morfoagronômicos para produtividade de grãos em feijoeiro comum

In Brazil, the common bean crop has representative agricultural exploitation, not only because of its production economic value, but also because there is a large cultivation area. This research aimed to investigate the direct and indirect relationship of morphological components on grain yield in common bean plants. This study was carried out in a Quartzarenic Neosol in the municipality of Cassilândia, Mato Grosso do Sul State, Brazil, in the agricultural year of 2016/2017. The evaluated traits were: grains yield (GY) with its primary components; mass of one hundred grains (HG); number of grains per plant (GP); number of grains per pod (GPP); dry weight of aerial parts (DWA); number of pod per plant (PP); plant dry mass (DM); plant high (PH); and stem diameter (SD). Initially, the Pearson's correlation among these traits was estimated and the correlation network was used to graphically express the obtained results. Analysis of these data through the statistical techniques of multicollinearity diagnosis followed by path analysis enabled to verify that the number of pod per plant, the mass of one hundred grains, and the number of grains per plant, among the primary components of grain yield, are the traits of greater potential to select and identify superior genotypes for grain productivity yield, and that dry matter and stem diameter traits showed a negative correlation with grain yield in common bean grains.

No Brasil, a cultura do feijoeiro tem uma exploração agrícola representativa, não só por seu valor econômico de produção, mas também porque existe grande área de cultivo. Este trabalho teve como objetivo investigar a relação direta e indireta de componentes morfológicos no rendimento de grãos nas plantas de feijão comum. Este estudo foi realizado em Neossolo Quartzarênico no município de Cassilândia, Estado de Mato Grosso do Sul, no ano agrícola de 2016/2017. As características avaliadas foram: produtividade de grãos (GY) com seus componentes primários; massa de cem grãos (HG); número de grãos por planta (GP); número de grãos por vagem (GPP); massa seca da parte aérea (DWA); número de vagens por planta (PP); massa seca da planta (DM); altura de planta (PH); e diâmetro do caule (SD). Inicialmente, a correlação de Pearson entre esses caracteres foi estimada e a rede de correlação foi usada para expressar graficamente os resultados obtidos. A análise desses dados através do diagnóstico de multicolinearidade seguida de análise de trilha permitiu verificar que o número de vagens por planta, a massa de cem grãos e o número de grãos por planta, entre os principais componentes do rendimento de grãos, são os caracteres de maior potencial para selecionar e identificar genótipos superiores para a produtividade de grãos no feijoeiro e que a matéria seca e o diâmetro do caule mostraram uma correlação negativa com a produtividade de grãos em grãos.

Molecular diagnosis of symptomatic toxoplasmosis: a 9-year retrospective and prospective study in a referral laboratory in São Paulo, Brazil

ABSTRACT Symptomatic forms of toxoplasmosis are a serious public health problem and occur in around 10-20% of the infected people. Aiming to improve the molecular diagnosis of symptomatic toxoplasmosis in Brazilian patients, this study evaluated the performance of real time PCR testing two primer sets (B1 and REP-529) in detecting Toxoplasma gondii DNA. The methodology was assayed in 807 clinical samples with known clinical diagnosis, ELISA, and conventional PCR results in a 9-year period. All samples were from patients with clinical suspicion of several features of toxoplasmosis. According to the minimum detection limit curve (in CT), REP-529 had greater sensitivity to detect T. gondii DNA than B1. Both primer sets were retrospectively evaluated using 515 DNA from different clinical samples. The 122 patients without toxoplasmosis provided high specificity (REP-529, 99.2% and B1, 100%). From the 393 samples with positive ELISA, 146 had clinical diagnosis of toxoplasmosis and positive conventional PCR. REP-529 and B1 sensitivities were 95.9% and 83.6%, respectively. Comparison of REP-529 and B1 performances was further analyzed prospectively in 292 samples. Thus, from a total of 807 DNA analyzed, 217 (26.89%) had positive PCR with, at least one primer set and symptomatic toxoplasmosis confirmed by clinical diagnosis. REP-529 was positive in 97.23%, whereas B1 amplified only 78.80%. After comparing several samples in a Brazilian referral laboratory, this study concluded that REP-529 primer set had better performance than B1 one. These observations were based after using cases with defined clinical diagnosis, ELISA, and conventional PCR.