Water strategy improves the inflorescence primordia formation of 'Arra 15' grapevine in the Brazilian semiarid region.

Failure in irrigation management of grapevines grown in the Brazilian semiarid region can affect bud fertility. Adequate irrigation, considering both the development of bunches in the current cycle and the formation of fertile buds for subsequent cycles, can bring significant advances to viticulture. Therefore, the objective of this research was to investigate the effect of different irrigation levels during flowering on the formation of buds and potential bunches of 'Arra 15' grapevine and its relationship with metabolic processes. A field experiment was carried out in a commercial vineyard in Petrolina, Pernambuco, Brazil, during the 2021 and 2022 seasons. The experiment was designed in randomized blocks with four replications and five irrigation levels (70; 85; 100; 115 and 130% of crop evapotranspiration - ETc) during three production cycles. The variables fertile bud, vegetative bud, dead bud, potential fertility of the basal, median, and apical regions of the branches, number of potential bunches, reducing sugar, total soluble sugar, net photosynthesis, stomatal conductance, transpiration, and relative chlorophyll index were evaluated. The 115% ETc irrigation level improved the number of fertile buds and number of potential bunches. Irrigation level above 115% ETc increased gas exchange and relative chlorophyll index, while 70% ETc increased leaf sugar content. The most appropriate irrigation strategy is the application of 115% ETc during the flowering stage, for the increase of fertile buds and potential bunches of the next cycle, without influencing the vine metabolism. Total soluble sugars are a promising indicator of water deficit during flowering and as an indicator of vegetative bud formation for the next cycle.

Extracellular Vesicles from Leishmania (Leishmania) infantum Contribute in Stimulating Immune Response and Immunosuppression in Hosts with Visceral Leishmaniasis.

Visceral leishmaniasis (VL) is a chronic systemic disease. In Brazil this infection is caused by Leishmania (Leishmania) infantum. Extracellular vesicles (EVs) released by Leishmania species have different functions like the modulation of host immune systems and inflammatory responses, among others. This study evaluated the participation of EVs from L. (L.) infantum (Leish-EVs) in recognition of the humoral and cellular immune response of hosts with VL. Promastigotes were cultivated in 199 medium and, in the log phase of growth, they were centrifuged, washed, resus-pended in RPMI medium, and incubated for 2 to 24 h, at 25 °C or 37 °C to release Leish-EVs. This dynamic was evaluated using transmission (TEM) and scanning (SEM) electron microscopies, as well as nanoparticle tracking analysis (NTA). The results suggested that parasite penetration in mammal macrophages requires more Leish-EVs than those living in insect vectors, since promastigotes incubated at 37 °C released more Leish-EVs than those incubated at 25 °C. Infected THP-1 cells produced high EV concentration (THP-1 cells-EVs) when compared with those from the control group. The same results were obtained when THP-1 cells were treated with Leish-EVs or a crude Leishmania antigen. These data indicated that host-EV concentrations could be used to distinguish infected from uninfected hosts. THP-1 cells treated with Leish-EVs expressed more IL-12 than control THP-1 cells, but were unable to express IFN-γ. These same cells highly expressed IL-10, which inhibited TNF-α and IL-6. Equally, THP-1 cells treated with Leish-EVs up-expressed miR-21-5p and miR-146a-5p. In conclusion, THP-1 cells treated with Leish-EVs highly expressed miR-21-5p and miR-146a-5p and caused the dysregulation of IL-10. Indirectly, these results suggest that high expression of these miRNAs species is caused by Leish-EVs. Consequently, this molecular via can contribute to immunosuppression causing enhanced immunopathology in infected hosts.

Identification and validation of reference genes of circulating microRNAs for use as control in gestational toxoplasmosis.

Toxoplasmosis causes serious harm to the fetus, as tachyzoite dissemination, during pregnancy in women developing the primo-infection. The microRNAs (miRNAs) are small non-coding RNAs, which have regulatory roles in cells by silencing messenger RNA. Circulating miRNA are promising biomarkers for diagnosis and prognosis of numerous diseases. The miRNAs levels are estimated by quantitative real-time PCR (qPCR), however, the relative quantification of each miRNA expression requires proper normalization methods using endogenous miRNAs as control. This study analyzed the expression of three endogenous miRNAs (miR-484, miR -423-3p and miR-26b-5p) for use as normalizers in future studies of target miRNAs for gestational toxoplasmosis (GT). A total of 32 plasma samples were used in all assays divided in 21 from women with GT and 11 from healthy women. The stability of each endogenous miRNA was evaluated by the algorithm methods RefFinder that included GeNorm, Normfinder, BestKeeper and comparative delta-CT programs. The miR-484 was the most stably gene, and equivalently expressed in GT and NC groups. These results contribute to future studies of target miRNAs in clinical samples of women with gestational toxoplasmosis.

Dogs with canine visceral leishmaniasis have a boost of extracellular vesicles and miR-21-5p up-expression.

This retrospective cohort study analysed extracellular vesicles (EVs) and microRNAs (miRNAs) excreted in canine sera from dogs with canine visceral leishmaniasis (CanVL). A total of 56 canine sera were divided into Group I (28, from healthy dogs) and Group II (28, from the same dogs, but already with CanVL). CanVL was determined by clinical and laboratory diagnoses. Canine sera were ultra-centrifuged to recover EVs (Can-EVs). Analyses by transmission electron microscopy, nanoparticle tracking analysis (NTA), sodium dodecyl sulfate-poli-acrylammide gel eletroforesis (SDS-PAGE) and, Immunoblot confirmed the presence of (i) microvesicles/exosomes and (ii) the tetraspanins CD63 and CD9. EVs secreted by Leishmania (Leishmania) infantum-EVs were reactive against sera from dogs with CanVL (performed by ELISA and Immunoblot). NTA analyses exhibited that concentrations of Can-EVs from dogs with CanVL (7.78 × 1010 Can-EVs/mL) were higher (p < .0001) than the non-infected dogs (mean: 1.47 × 1010 Can-EVs/mL). These results suggested that concentrations of Can-EVs were able to distinguish dogs with CanVL from healthy dogs. The relative expressions of 11 miRNAs species (miR-21-5p, miR-146a-5p, miR-125b-5p, miR-144-3p, miR-194-5p, miR-346, miR-29c-3p, miR-155-5p, miR-24-3p, miR-181a-5p, and miR-9-5p) were estimated in purified miRNAs of 30 canine sera. Dogs with CanVL up-expressed miR-21-5p and miR-146a-5p when compared with healthy dogs. The other miRNA species were poorly or not expressed in canine sera. In conclusion, this study suggests that CanVL induces changes in size and concentration of Can-EVs, as well as, the up-expression of miR-21-5p and miR-146a-5p in infected dogs.

Developing an algorithm to geographically estimate the available time for agricultural field spraying in the state of Mato Grosso do Sul / Desenvolvendo um algoritmo para estimar geograficamente o tempo disponível para pulverização agrícola no estado de Mato Grosso do Su

The dimensions of mechanized agricultural systems depend on the edaphoclimatic conditions, crops, and work regimes. This study aimed to geographically estimate the monthly available time and number of favorable hours for agricultural field spraying in the state of Mato Grosso do Sul, Brazil. The meteorological restrictions imposed during unfavorable hours were as follows: ambient temperature above 32 ºC, relative humidity below 50 %, wind speed above 15 km h-1, and volumetric soil humidity above 39 % (equivalent to 90 % of the available water capacity). Mathematical models were then developed considering a period of ten years, which used historical data from the ground monitoring stations of the National Institute of Meteorology within the region. The subsequent algorithm was programmed and installed in a web server to simulate the time required for agricultural field spraying. During the cropping period in the region, there were climatic restrictions on performing agricultural spraying, with relative humidity being the variable with the most significant impact. However, soil moisture conditions restricted the available time for agricultural spraying more than the wind speed, relative air humidity, or ambient temperature.

O dimensionamento de sistemas mecanizados agrícolas depende das condições edafoclimáticas, da cultura e do regime de trabalho. O objetivo deste trabalho foi estimar geograficamente o tempo disponível mensal e o número de horas favoráveis à pulverização agrícola no Estado de Mato Grosso do Sul, Brasil. As restrições meteorológicas impostas para o cálculo das horas desfavoráveis foram: temperatura ambiente acima de 32ºC, umidade relativa abaixo de 50%, velocidade do vento acima de 15 km h-1 e umidade volumétrica do solo acima de 39% (equivalente a 90% da capacidade hídrica disponível). Os modelos matemáticos foram elaborados considerando um período de dez anos, com base nos dados históricos das estações automáticas do Instituto Nacional de Meteorologia instaladas no Estado. O algoritmo desenvolvido e instalado em ambiente web permitiu simular o tempo disponível para pulverização agrícola no estado de Mato Grosso do Sul. Durante o período de safra na região, há restrições climáticas para a realização da atividade de pulverização agrícola, sendo a umidade relativa do ar a variável de maior impacto. A condição de umidade do solo é mais restritiva para o tempo disponível para a pulverização agrícola do que a velocidade do vento, umidade relativa do ar ou temperatura do ambiente.

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.

Performance of a Real Time PCR for Pneumocystis jirovecii Identification in Induced Sputum of AIDS Patients: Differentiation between Pneumonia and Colonization.

Pneumocystis jirovecii pneumonia (PcP) remains an important cause of morbimortality worldwide and a diagnostic challenge. Conventional methods have low accuracy, hardly discriminating colonization from infection, while some new high-cost or broncho-alveolar lavage-based methods have limited usefulness in developing countries. Quantitative PCR (qPCR) tests may overcome these limitations due to their high accuracy, possibility of automation, and decreasing cost. We evaluated an in-house qPCR targeting the fungus mtSSU gene using induced sputum. Sensitivity of the assay (ten target gene copies/assay) was determined using recombinant plasmids. We prospectively studied 86 AIDS patients with subacute respiratory symptoms in whom PcP was suspected. qPCR results were determined as quantification cycles (Cq) and compared with a qualitative PCR performed in the same IS, serum 1,3-ß-D-Glucan assay, and a clinical/laboratory/radiology index for PcP. The qPCR clustered the patients in three groups: 32 with Cq ≤ 31 (qPCR+), 45 with Cq ≥ 33 (qPCR-), and nine with Cq between 31-33 (intermediary), which, combined with the other three analyses, enabled us to classify the groups as having PcP, not P. jirovecii-infected, and P. jirovecii-colonized, respectively. This molecular assay may contribute to improve PcP management, avoiding unnecessary treatments, and our knowledge of the natural history of this infection.

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).

Ocular Disease in Mice Inoculated with Pork Heart Samples Infected with Toxoplasma gondii.

PURPOSE: We analyzed the frequency, viability, and genetic characteristics of T. gondii in pork heart samples. METHODS: Thirty-five fresh pork samples were purchased in a slaughterhouse in Erechim city. The DNA was extracted and qPCR was performed. T. gondii genotyping was performed using PCR-RFLP analysis. Positive samples were digested and inoculated in mice for viability analysis. RESULTS: Our results showed that T. gondii DNA was detected in 25.7% of the pork heart samples and genotyping revealed one new atypical strain. The viability analyses demonstrated that 40% of mice presented clinical signs of T. gondii infection. qPCR was positive in the lung, liver, and brain of mice that presented clinical signs of T. gondii infection. Also, the histopathology analysis showed retinal disorganization, retinal detachment, inflammatory cell infiltration, and fibrosis in the eyes analyzed. CONCLUSION: Our findings have shown that pork eat from southern Brazil may contain live T. gondii that could be associated with toxoplasmosis.

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.

Combining urine antigen and blood polymerase chain reaction for the diagnosis of disseminated histoplasmosis in hospitalized patients with advanced HIV disease.

Disseminated histoplasmosis (DH) is endemic in Latin America and the Caribbean where diagnostic tools are restricted. We carried-out a 1-year prospective cohort study at a referral hospital in São Paulo, Brazil. Participants had > or =18 years old, were hospitalized due to any indication and had CD4+ < 200 cells/µl. A urine commercial monoclonal Histoplasma galactomannan enzyme-linked immunosorbent assay (IMMY, Norman, OK, USA) and 'in house' Histoplasma blood nested PCR were performed in all cases. Probable/proven DH cases were defined according to international guidelines. Conventional mycological methods were available in routine conditions to investigate suspected DH cases. Treatment of participants followed the institutional routine. One-hundred six participants were included. Median age (interquartile range [IQR]) was 39.5 years (30.0-47.3) and 80 individuals (75.5%) were males. Median (IQR) CD4 cell count was 26.5 (9.4-89.3) cells/mm3. DH was diagnosed in 8/106 patients (7.5%). Antigen assay and/or PCR were positive in 4.7% (5/106) of patients. The antigen assay and/or PCR identified 37.5% (3/8) of DH cases, which had not been diagnosed with conventional mycological methods, but had clinical manifestations compatible with HD. In conclusion, the use of Histoplasma urine antigen and Histoplasma blood PCR guided by CD4 status contributed to the diagnosis of DH in hospitalized individuals. These assays were complementary to conventional mycologic methods and are urgently needed in our setting. LAY SUMMARY: In this prospective cohort study carried-out in a referral center in São Paulo, Brazil, we found a high frequency of AIDS-related disseminated histoplasmosis (8/106, 7.5%). We used urine antigen test and blood PCR assay to improve the diagnosis of this opportunistic disease.

Increase in the irrigated area of sugarcane and its potential in the bioenergetic generation of Brazil / Aumento da área irrigada de cana-de-açúcar e seu potencial na geração bioenergética do Brasil

Sugarcane has alarge capacity for producing electricity using completely clean technology from a renewable source, contributing to environmental preservation. This study aimed to evaluate the increase in biomass resulting from the implementation of irrigation in Brazilian sugarcane plantations, estimate the potential for cogeneration, and calculate how much it will be possible to increase the Brazilian energy cogenerated with sugarcane bagasse and the impact on the Brazilian energy matrix. The increase in irrigation in the rainfed areas of Brazilian sugarcane plantations has substantial potential in increasing biomass for energy cogeneration. Considering an increase of 15% in the sugarcane plantation yields in rainfed areas achieved by the increase in irrigation, it leadsto a potential increase of 96.39 million tons of sugarcane and 26.80 million tons biomass produced. In the energy matrix, the potential impact is 1.42 GWh of cogenerated energy, which corresponds to a 12.47% increase in cogeneration in the sugar-energy sector and a 0.85% increase in Brazil's energy matrix. The expansion of the sugarcane irrigated area contributes to the increase in the production of bagasse and bioenergetic generation in Brazil.

A cana-de-açúcar tem grande capacidade de produção de energia elétrica com tecnologia totalmente limpa, de fonte renovável, contribuindo com a preservação ambiental. O objetivo da presente pesquisa foi avaliar o incremento de biomassa advinda da implantação de irrigação nos canaviais brasileiros, estimar o potencial de cogeração, e calcular o quanto será possível aumentar a energia brasileira cogerada com bagaço de cana-de-açúcar e o impacto na matriz energética brasileira. O incremento de irrigação nas áreas de sequeiros dos canaviais brasileiros tem um potencial substancial no incremento de biomassa para a cogeração de energia.Considerando um aumento de 15% na produção dos canaviais das áreas de sequeiro, conquistados a partir do incremento de irrigação, ocasiona em um potencial de aumento de 96,39 milhões de toneladas de cana-de-açúcar e 26,80 milhões de toneladas de biomassa produzida. Na matriz energética, o impacto potencial é de 1,42 GWh de energia cogerada que corresponde a 12,47% de aumento de cogeração no setor sucroenergético e 0,85% de aumento na matriz energética do Brasil. A expansão da área irrigada de cana-de-açúcar contribui com o aumento da produção de bagaço e geração bioenergética do Brasil.

Increase in the irrigated area of sugarcane and its potential in the bioenergetic generation of Brazil / Aumento da área irrigada de cana-de-açúcar e seu potencial na geração bioenergética do Brasil

Sugarcane has alarge capacity for producing electricity using completely clean technology from a renewable source, contributing to environmental preservation. This study aimed to evaluate the increase in biomass resulting from the implementation of irrigation in Brazilian sugarcane plantations, estimate the potential for cogeneration, and calculate how much it will be possible to increase the Brazilian energy cogenerated with sugarcane bagasse and the impact on the Brazilian energy matrix. The increase in irrigation in the rainfed areas of Brazilian sugarcane plantations has substantial potential in increasing biomass for energy cogeneration. Considering an increase of 15% in the sugarcane plantation yields in rainfed areas achieved by the increase in irrigation, it leadsto a potential increase of 96.39 million tons of sugarcane and 26.80 million tons biomass produced. In the energy matrix, the potential impact is 1.42 GWh of cogenerated energy, which corresponds to a 12.47% increase in cogeneration in the sugar-energy sector and a 0.85% increase in Brazil's energy matrix. The expansion of the sugarcane irrigated area contributes to the increase in the production of bagasse and bioenergetic generation in Brazil.(AU)

A cana-de-açúcar tem grande capacidade de produção de energia elétrica com tecnologia totalmente limpa, de fonte renovável, contribuindo com a preservação ambiental. O objetivo da presente pesquisa foi avaliar o incremento de biomassa advinda da implantação de irrigação nos canaviais brasileiros, estimar o potencial de cogeração, e calcular o quanto será possível aumentar a energia brasileira cogerada com bagaço de cana-de-açúcar e o impacto na matriz energética brasileira. O incremento de irrigação nas áreas de sequeiros dos canaviais brasileiros tem um potencial substancial no incremento de biomassa para a cogeração de energia.Considerando um aumento de 15% na produção dos canaviais das áreas de sequeiro, conquistados a partir do incremento de irrigação, ocasiona em um potencial de aumento de 96,39 milhões de toneladas de cana-de-açúcar e 26,80 milhões de toneladas de biomassa produzida. Na matriz energética, o impacto potencial é de 1,42 GWh de energia cogerada que corresponde a 12,47% de aumento de cogeração no setor sucroenergético e 0,85% de aumento na matriz energética do Brasil. A expansão da área irrigada de cana-de-açúcar contribui com o aumento da produção de bagaço e geração bioenergética do Brasil.(AU)

Angiostrongylus cantonensis in urban populations of terrestrial gastropods and rats in an impoverished region of Brazil.

The nematode Angiostrongylus cantonensis is the most common cause of neuroangiostrongyliasis (manifested as eosinophilic meningitis) in humans. Gastropod molluscs are used as intermediate hosts and rats of various species are definitive hosts of this parasite. In this study, we identified several environmental factors associated with the presence and abundance of terrestrial gastropods in an impoverished urban region in Brazil. We also found that body condition, age and presence of co-infection with other parasite species in urban Rattus norvegicus, as well as environmental factors were associated with the probability and intensity of A. cantonensis infection. The study area was also found to have a moderate prevalence of the nematode in rodents (33% of 168 individuals). Eight species of molluscs (577 individuals) were identified, four of which were positive for A. cantonensis. Our study indicates that the environmental conditions of poor urban areas (presence of running and standing water, sewage, humidity and accumulated rain and accumulation of construction materials) influenced both the distribution and abundance of terrestrial gastropods, as well as infected rats, contributing to the maintenance of the A. cantonensis transmission cycle in the area. Besides neuroangiostrongyliasis, the presence of these hosts may also contribute to susceptibility to other zoonoses.

Immunization with extracellular vesicles excreted by Toxoplasma gondii confers protection in murine infection, activating cellular and humoral responses.

The study aim was to analyze whether microvesicles and exosomes, named extracellular vesicles (EVs), purified from Toxoplasma gondii are able to stimulate the protective immunity of experimental mice when administered, as challenge, a highly virulent strain. EVs excreted from T. gondii tachyzoites (RH strain) were purified by chromatography and used for immunization assays in inbred mouse groups (EV-IM). Chronic infected (CHR) and naive (NI) mice were used as control groups, since the immune response is well known. After immunizations, experimental groups were challenged with 100 tachyzoites. Next, parasitemias were determined by real-time PCR (qPCR), and survival levels were evaluated daily. The humoral response was analyzed by detection of IgM, IgG, IgG1 and IgG2a, and opsonization experiments. The cellular response was evaluated in situ by immunohistochemistry on IFN-γ, IL-10, TNF-α and IL-17 expression in cells of five organs (brain, heart, liver, spleen and skeletal muscles). EV immunization reduced parasitemia and increased the survival index in two mouse lineages (A/Sn and BALB/c) infected with a lethal T. gondii strain. EV-IM mice had higher IgG1 levels than IgM or IgG2a. IgGs purified from sera of EV-IM mice were able to opsonize tachyzoites (RH strain), and mice that received these parasites had lower parasitemias, and mortality was delayed 48 h, compared with the same results from those receiving parasites opsonized with IgG purified from NI mice. Brain and spleen cells from EV-IM mice more highly expressed IFN-γ, IL-10 and TNF-α. In conclusion, EV-immunization was capable of inducing immune protection, eliciting high production of IgG1, IFN-γ, IL-10 and TNF-α.

First record of natural infection by Angiostrongylus cantonensis (Nematoda: Metastrongyloidea) in Belocaulus willibaldoi and Rattus norvegicus in an urban area of São Paulo city, SP, Brazil.

Angiostrongylus cantonensis, a rat lungworm, is one of the leading causes of eosinophilic meningitis in humans. Infection in humans occurs by the ingestion of intermediate hosts, undercooked paratenic hosts or contaminated vegetables and fruits by mucus from infected molluscs. This zoonosis is widespread in tropical and subtropical areas of Southeast Asia, it has also been reported in the Pacific Islands as well as in other regions of Americas. In Brazil, human cases of angiostrongyliasis have been reported since 2007 in Southeast, Northeast, and South regions. In January 2011, we collected a batch of 30 Belocaulus willibaldoi slides in a neighborhood of São Paulo city (Parque Fernanda). Six of them were used for identifying species, and the others (24) were used in parasitological tests through digestion in peptic solution and then larvae isolation by the Rugai method. A total of 250 larvae were obtained and they had morphological traits of Angiostrongylus spp. Later, four Golden hamsters (Mesocricetus auratus) were infected with 38 larvae that allowed the recovery of young worms from the brain and lungs of rodents on the 21st and 30th day of infection. In this same neighborhood we captured rodents (Rattus norvegicus) that, after necropsy led us to recovery of 22 adult worms in the pulmonary arteries (14 males and 8 females) in May 2011. The larvae and worms obtained from natural infection were evaluated by morphological and morphometric parameters, as well as biological behavior patterns and molecular profile. All methodologies identified the parasite as Angiostrongylus cantonensis. In this way, we report for the first time, the natural infection by A. cantonensis in intermediate (B. willibaldoi) and definitive (R. norvegicus) hosts in a new urban region of Brazil.

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.

Plasma extracellular microRNAs are related to AIDS/cerebral toxoplasmosis co-infection

This study investigated the potential of five miRNA candidates for cerebral toxoplasmosis/HIV co-infection (CT/HIV) biomarkers. miR-155-5p, miR-146a-5p, miR-21-5p, miR-125b-5p and miR-29c-3p were tested in 79 plasma divided into groups: 32 CT/HIV patients; 27 individuals with asymptomatic toxoplasmosis (AT); and 20 individuals seronegative for toxoplasmosis (NC). From each was collected peripheral blood/EDTA for laboratory diagnosis. Blood cells for DNA extractions (molecular diagnosis), plasma for RNA extractions (gene expression) and ELISA (serological diagnosis). miRNA expression was performed by qPCR, and values were expressed in Relative Quantification (RQ). Among the five miRNAs, miR-21-5p and miR-146a-5p were up-expressed in CT/HIV group when compared with AT and NC groups. RQ means for miR-21-5p and miR-146a-5p in CT/HIV group were 3.829 and 2.500, while in AT group, were 1.815 and 1.661, respectively. Differences between 3 groups were statistically significant (Kruskal-Wallis ANOVA test), as well as CT/HIV and AT groups (Mann-Whitney test). Plasma of CT/HIV and AT groups expressed similar levels of miR-29c-3p, miR-155-5p and miR-125b-5p. As NC group was different of CT/HIV and AT groups, differences between three groups were statistically significant (Kruskal-Wallis ANOVA test). No difference was shown between CT/HIV and AT groups (Mann-Whitney test). These results suggest the host miRNAs modulation by Toxoplasma gondii

A new strain of Toxoplasma gondii circulating in southern Brazil.

Recently, an outbreak of human toxoplasmosis was identified in Santa Maria city, Southern Brazil. However, the suspected vehicle of Toxoplasma gondii contamination in this region remains unclear. This study was conduct to analyze whether pork meat samples collected in supermarkets from Santa Maria city, RS, could be infected with T. gondii. Thus, we analyzed the presence of T. gondii DNA in 20 pork hearts, 20 pork tongues and 20 sausages. DNA was extracted from each sample and real-time PCR was performed using 529-bp and B1 markers. T. gondii genotyping was performed by PCR-RFLP analysis. T. gondii DNA was detected in 2 of 20 (10%) heart samples and in 1 of 20 (5%) tongue samples using 529-bp marker. Besides, those 2 (10%) heart samples also were positive for T. gondii using B1 marker. All sausage samples were negative for both markers. Genotyping revealed a new atypical genotype in the pork meat. Our findings were not able to confirm whether these food samples were involved in some outbreak. However, we can conclude that food samples containing T. gondii can be displayed in Santa Maria supermarkets. In addition, a new T. gondii genotype was identified circulating in southern Brazil.