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Efficient and sustainable sludge management is a significant environmental and health challenge. Sludge-treatment reed beds (STRBs) are widely recognized as a cost-effective, highly efficient, and environmentally friendly solution for sludge treatment and dewatering. This study investigated the bacterial community composition and diversity in pilot-scale STRBs operating at different sludge loading rates (75, 100, and 125 ​kg ​m2 year−1). 16S rRNA V4 DNA sequencing was used to assess the diversity of the bacterial communities within the sludge samples. The relative abundance of prokaryotic taxa was affected by all treatments. As the sludge loads increased, the Shannon entropy and evenness diversity also increased for the STRBs and unplanted beds. Interestingly, the presence of reeds resulted in significantly lower Shannon and evenness indices than unplanted beds, regardless of the sludge. Additionally, the correlation network analysis revealed distinct microbial clusters with distinct responses to reeds and sludge loads. Principal component analysis evidenced an association between cluster 5 and organic matter decomposition, primarily at higher sludge doses, while clusters 4 and 6 were related to sludge decomposition at lower doses. Additionally, cluster 4 was associated with nutrient removal. The formation of distinct microbial niches was linked to sludge stabilization and nutrient removal and was influenced by both sludge loading rates and the presence of reeds. Future research can leverage these findings to innovate pollutant removal and ecosystem services for sludge treatment, thus advancing sustainable sludge management and environmental preservation.

Este texto tem como objetivo compartilhar parte das experiências vivenciadas no curso de extensão online Ritmos Brasileiros para Violão, realizado em quatro edições entre os anos de 2020 e 2022, como iniciativa de três docentes do curso de Graduação em Música da Universidade Federal de Uberlândia. Pretende-se, aqui, fazer um breve histórico das ações propostas, refletir sobre a forma de elaboração do projeto e seu impacto junto aos participantes. O presente relato também serve como forma de avaliar o trabalho e de se pensar em possíveis desdobramentos da atividade extensionista em um momento pós-pandemia.

By 2050, the global population is projected to reach 9 [...]

Long term intensive agricultural farming has impacted negatively the soil health, especially in arid climates. This study aimed to investigate the impact of long-term (1993 to 2020) farming practices on the chemical and biological properties of soil and water of Al-Batinah region. Soil and water samples were collected from eight farms and characterized for their chemical and biological soil properties. The results of soil samples indicated significant (p ≤ 0.05) changes in average soil chemical and biological properties over time. Soil electrical conductivity (ECe) increased from 'low salinity' (1.5 dS m−1 ± 0.49) to 'highly saline' (6.6 dS m−1 ± 1.35), and soil pH decreased from 8.4 to 7.5. The organic matter increased from 'very low' (0.2%) to 'medium' (1.9%), and the Olsen phosphorus increased from deficient (6.5 mg kg−1 ± 0.9) to excessive (76 mg kg−1 ± 10.4). Sodium, magnesium, chloride, and sulfate ions were also significantly increased during this period, while no significant differences was observed in soil potassium, and calcium concentration. The soil (ECe) showed significant positive correlation with soil sodium, magnesium, chloride, sulfate, potassium, calcium, and water electrical conductivity (EC), nitrate, sodium, potassium, and significant negative correlation with soil pH. Soil organic matter showed significant a positive correlation with soil phosphorus. Soil sodium showed significant positive correlations with soil magnesium, chloride, sulfate, potassium, calcium, and water electrical conductivity (EC), nitrate, sodium, and potassium. The Al-Batinah region soils showed some positive changes including an increase in organic matter, and a decrease in soil pH. Also, negative changes occurred such an excessive increase of soil phosphorus, sodium, magnesium, chloride, and sulfate ions. Our findings can be used to inform the public policies and strategies for a more sustainable soil and agricultural management in Oman..

Por meio deste estudo teórico, é possível constatar que a Internet, criada inicialmente como uma rede que possibilitaria o acesso e a descentralização da circulação de informação, a viabilização da comunicação e, consequentemente, o acesso de todos às plataformas de informação, negócios, serviços e redes sociais online, não é uma rede aleatória, e as suas propriedades de funcionamento nos obrigam a reavaliar a ideia geral, segundo a qual a rede seria um espaço de liberdade de expressão, justiça, democracia e transformação social. Atualmente são as grandes empresas de tecnologia digital quem têm dominado a arquitetura da rede e ditado a maneira como esta deve funcionar, ocupando, assim, um papel econômico e político em nossa sociedade. Isso porque a grande maioria dos serviços públicos e privados, assim com a vida de cada um, tem sido atravessada pelo digital, o que provocava mudanças nas organizações sociais, e consequentemente nos modos de subjetivação contemporânea.

Congenital Zika syndrome (CZS) is a set of birth defects caused by Zika virus (ZIKV) infection during pregnancy. Microcephaly is its main feature, but other brain abnormalities are found in CZS patients, such as ventriculomegaly, brain calcifications, and dysgenesis of the corpus callosum. Many studies have focused on microcephaly, but it remains unknown how ZIKV infection leads to callosal malformation. To tackle this issue, we infected mouse embryos in utero with a Brazilian ZIKV isolate and found that they were born with a reduction in callosal area and density of callosal neurons. ZIKV infection also causes a density reduction in PH3+ cells, intermediate progenitor cells, and SATB2+ neurons. Moreover, axonal tracing revealed that callosal axons are reduced and misrouted. Also, ZIKV-infected cultures show a reduction in callosal axon length. GFAP labeling showed that an in utero infection compromises glial cells responsible for midline axon guidance. In sum, we showed that ZIKV infection impairs critical steps of corpus callosum formation by disrupting not only neurogenesis, but also axon guidance and growth across the midline.

Leishmania amazonensis can cause cutaneous and visceral clinical manifestations of leishmaniasis in infected hosts. Once the treatment against disease is toxic, presents high cost, and/or there is the emergence of parasite-resistant strains, alternative means through which to control the disease must be developed. In this context, immunotherapeutics combining known drugs with immunogens could be applied to control infections and allow hosts to recover from the disease. In this study, immunotherapeutics protocols associating mimotopes selected by phage display and amphotericin B (AmpB) were evaluated in L. amazonensis-infected mice. Immunogens, A4 and A8 phages, were administered alone or associated with AmpB. Other animals received saline, AmpB, a wild-type phage (WTP), or WTP/AmpB as controls. Evaluations performed one and thirty days after the application of immunotherapeutics showed that the A4/AmpB and A8/AmpB combinations induced the most polarized Th1-type immune responses, which reflected in significant reductions in the lesion’s average diameter and in the parasite load in the infected tissue and distinct organs of the animals. In addition, the combination also reduced the drug toxicity, as compared to values found using it alone. In this context, preliminary data presented here suggest the potential to associate A4 and A8 phages with AmpB to be applied in future studies for treatment against leishmaniasis.

A Casca da Castanha de Caju (CCC) é uma biomassa residual proveniente do processo de beneficiamento da castanha de caju e o uso adequado desse resíduo vem se tornando cada vez mais atrativo. Desta forma, o presente trabalho possui como objetivo aplicar a metodologia da Avaliação do Ciclo de Vida (ACV) para compreender os impactos ambientais desses processos, em unidades-piloto com capacidade de processamento de 150 kg/h, visando à geração de energia. Uma ACV do tipo gate-to-gate (fronteiras do sistema) foi aplicada. Na categoria GWP (Potencial de Aquecimento Global) foi verificado que a queima do carvão oriundo da pirólise emite anualmente 114 tCO2-eq e a gaseificação apenas 80,4 tCO2-eq. A queima do syngas no gerador, em ambos os processos, foi responsável pelas emissões de NOX que sensibilizaram a categoria POCP (Potencial de formação de Ozonio Fotoquimico). Já na categoria de emissão de PM2,5 (Material Particulado de 2,5 m) o carvão da pirólise mais uma vez foi um dos maiores responsáveis pelo agravamento dos impactos ambientais desta categoria, emitindo um total de 21,9 kg de PM2,5- eq, enquanto que o processo de gaseificação foi responsável somente pela emissão de 0,25 % desse total.

Aquaponics are efficient systems that associate aquatic organisms’ production and plants by recirculating water and nutrients between aquaculture and hydroponic tanks. In this study, we characterised the bacterial communities in the freshwater aquaponics system that can mineralise polysaccharides and phytate by producing carbohydrate-degrading enzymes and phytases, by 16S rRNA gene sequencing and in vitro culture techniques. Around 20% of the operational taxonomic units (zOTUs) identified were previously reported to carry fibre-degrading enzyme putative genes, namely β-glucanase (1%), xylanase (5%), or cellulases (17%). Ten % of the zOTUs were previously reported to carry putative genes of phytases with different catalytic mechanisms, namely β-propeller (6%), histidine acid phytases (3%), and protein tyrosine phytase (Staphylococcus sp. and Rossellomorea sp. could produce consistent extracellular phytate-degrading activity. The PCR amplification of β-propeller genes both in environmental samples and in the isolates obtained showed that this is the most ecologically relevant phytase type in the aquaponics systems used. In summary, the aquaponics system is abundant with bacteria carrying enzymes responsible for plant-nutrient mineralisation.

Abstract Background The development of a vaccine for the prevention of visceral leishmaniasis (VL) still represents a significant unmet medical need. A human vaccine can be found if one takes into consideration that many people living in endemic areas of disease are infected but do not develop active VL, including those subjects with subclinical or asymptomatic infection. Methods In this study, a phage display was used to select phage-exposed peptides that were specific to immunoglobulin G (IgG) antibodies from asymptomatic and symptomatic VL patients, separating them from non-infected subjects. Phage clones presenting valid peptide sequences were selected and used as stimuli of peripheral blood mononuclear cells (PBMCs) obtained from both patients’ groups and controls. Those with higher interferon-gamma (IFN-γ)/interleukin (IL)-10 ratios were further selected for vaccination tests. Results Among 17 evaluated clones, two were selected, B1 and D11, and used to immunize BALB/c mice in an attempt to further validate their in vivo protective efficacy against Leishmania infantum infection. Both clones induced partial protection against the parasite challenge, which was evidenced by the reduction of parasitism in the evaluated organs, a process mediated by a specific T helper (Th)1 immune response. Conclusions To the best of our knowledge, this study is the first to use a rational strategy based on in vitro stimulation of human PBMCs with selected phage-displayed clones to obtain new immunogens against VL.

Resumo No Seminário da ética, Lacan opôs a pastoral analítica à ética do “não ceder de seu desejo”. Ao pôr o desejo como parâmetro, tal ética questiona valores universais para alcançar uma suposta boa vida. Comparamos tal formulação à ética do “bem-dizer o desejo”, destacada por Lacan em Televisão. Se no Seminário 7 temos uma crítica aos ideais de felicidade associados ao mercado dos bens, na ética do “bem-dizer o desejo”, a felicidade aparece como parte do sintoma, vinculada aos diferentes modos de gozo, via possível para questionar a atual injunção a aproveitar, vinculada a uma certa cultura da alta performance.

Visceral leishmaniasis (VL) still represents a serious public health problem in Brazil due to the inefficiency of the control measures currently employed, that included early diagnosis and treatment of human cases, vector control, euthanasia of infected dogs and, recently approved in Brazil, treatment with Milteforam drug. Effective clinical management depend largely on early and unequivocal diagnosis, however, cross-reactivity have also been described in serological tests, especially when it refers to individuals from areas where Chagas' disease is also present. Thus, to discover new antigens to improve the current serological tests for VL diagnosis is urgently needed. Here, we performed an immunogenomic screen strategy to identify conserved linear B-cell epitopes in the predicted L. infantum proteome using the following criteria: i) proteins expressed in the stages found in the vertebrate host, amastigote stage, and secreted/excreted, to guarantee greater exposure to the immune system; ii) divergent from proteins present in other infectious disease pathogens with incidence in endemic areas for VL, as T. cruzi; iii) highly antigenic to humans with different genetic backgrounds, independently of the clinical stage of the disease; iv) stable and adaptable to quality-control tests to guarantee reproducibility; v) using statistical analysis to determine a suitable sample size to evaluate accuracy of diagnostic tests established by receiver operating characteristic strategy. We selected six predicted linear B-cell epitopes from three proteins of L. infantum parasite. The results demonstrated that a mixture of peptides (Mix IV: peptides 3+6) were able to identify VL cases and simultaneously able to discriminate infections caused by T. cruzi parasite with high accuracy (100.00%) and perfect agreement (Kappa index = 1.000) with direct methods performed by laboratories in Brazil. The results also demonstrated that peptide-6, Mix III (peptides 2+6) and I (peptides 2+3+6) are potential antigens able to used in VL diagnosis, represented by high accuracy (Ac = 99.52%, 99.52% and 98.56%, respectively). This study represents an interesting strategy for discovery new antigens applied to serologic diagnosis which will contribute to the improvement of the diagnosis of VL and, consequently, may help in the prevention, control and treatment of the disease in endemic areas of Brazil.

Abstract Background LiHyV is an antigenic hypothetical protein present in both promastigote and amastigote stages of Leishmania infantum, which was recently identified by an immunoproteomic approach. A recombinant version of this protein (rLiHyV) was evaluated as a diagnostic marker for canine VL (CVL). In addition, the prophylactic efficacy of the rLiHyV protein, and two of its CD8+ T cell epitopes, has been analyzed in a murine model of visceral leishmaniasis (VL). Methods Initially, the rLiHyV protein was evaluated by an ELISA technique for the serodiagnosis of CVL. Secondly, vaccines composed of the recombinant protein and both chemically synthesized peptides, combined with saponin as an adjuvant; were administered subcutaneously into BALB/c mice. The cellular and humoral responses generated by vaccination were evaluated. In addition, the parasite burden and immune response were studied 10 weeks after L. infantum infection. Results The rLiHyV protein was recognized by antibodies of VL dogs. No cross-reactivity was obtained with sera from dogs vaccinated with a Brazilian commercial vaccine, with sera from animals infected with Trypanosoma cruzi, Babesia canis and Ehrlichia canis, or those from non-infected animals living in an endemic area for leishmaniasis. After challenge with L. infantum, spleen cells of BALB/c mice vaccinated with rLiHyV/saponin stimulated with parasite antigens showed a higher production of IFN-γ, IL-12 and GM-CSF, than the same cells obtained from mice vaccinated with the individual peptides, or mice from control (inoculated with saline or saponin) groups. This Th1-type cellular response observed in rLiHyV/saponin vaccinated mice was accompanied by the induction of parasite-specific IgG2a isotype antibodies. Animals immunized with rLiHyV/saponin showed significant reductions in the parasite burden in the liver, spleen, bone marrow and in the lymph nodes draining the paws relative to control mice. Conclusions The present study showed for the first time that the L. infantum LiHyV protein could be considered as a vaccine candidate against L. infantum infection, as well as a diagnostic marker for CVL.

Abstract Background Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World. The pathology of the infection is determined by both host immune factors and species/strain differences of the parasite. Dogs represent the major reservoir of Leishmania infantum (syn. L. chagasi) and vaccines are considered the most cost-effective control tools for canine disease. Methods Selection of immunodominant peptides was performed by Phage Display to identify sequences recognized by L. infantum naturally infected animals. Sera from Leishmania infected animals were used in the biopanning to selection of specific peptides. Serum samples from T. cruzi infected and healthy animals were used as control. After selection, synthetic peptides were produced in membrane (spot-synthesis) in soluble form and blotting and ELISA were performed for validation of serum reactivity. Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice. Protection was determined by qPCR after challenge infection with virulent L. infantum. Results We reported the selection of Peptide 5 through Phage Display technique and demonstrate its ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier. Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection. Conclusion Peptide 5 is a promising vaccine candidate and the findings obtained in the present study encourage canine trials to confirm the effectiveness of a vaccine against CVL.

ABSTRACT The purpose of this study was to evaluate the efficiency of using total and partial analysis data of the trunk of Tectona grandis trees and of permanent plots, for the construction of site index curves and growth and production modeling, at the level of individual trees and of the settlement. Data were collected in settlements located in the State of Mato Grosso, with ages ranging from 2 to 10 years at the time of tree logging for trunk analysis. At that time, 40 permanent plots were installed, that were measured in the five subsequent years. Trunk analysis data obtained from those plots were used for validation of a variable density model. This model estimated the production with accuracy and consistency. Then, data from permanent plots were added to those of trunk analysis and the model was readjusted. It can be concluded that the trunk analysis of Tectona grandis trees is an efficient alternative of data collection for growth and production studies. The possibility of grouping past data, obtained by trunk analysis, with permanent plots, for growth modeling purposes, was also confirmed. Finally, the efficiency of trunk analysis data for growth modeling at a level of individual trees was evaluated and confirmed.

Neste artigo investigaremos a noção freudiana de sublimação e seu uso como mecanismo explicativo para os processos artísticos. Notaremos que, pela sua construção, aliás, confusa e carente de formalização conceitual, é impossível separar a sublimação do campo clínico no qual nasce e no qual encontra sua potência, como descrição de uma vicissitude e de uma possibilidade de lidar com a pulsão. Dessa forma, proporemos uma delimitação de qualquer "metapsicologia da arte" como instrumento clínico, antes que como instrumento de análise ou crítica da arte. Não obstante, notaremos a influência recíproca entre a psicanálise e a estética de seu tempo.

The conventional curriculum of Analytical Chemistry undergraduate courses emphasizes the introduction of techniques, methods and procedures used for instrumental analysis. All these concepts must be integrated into a sound conceptual framework to allow students to make appropriate decisions. Method calibration is one of the most critical parameters that has to be grasped since most analytical techniques depend on it for quantitative analysis. The conceptual understanding of calibration is not trivial for undergraduate students. External calibration is widely discussed during instrumental analysis courses. However, the understanding of the limitations of external calibration to correct some systematic errors is not directly derived from laboratory examples. The conceptual understanding of other calibration methods (standard addition, matrix matching, and internal standard) is imperative. The aim of this work is to present a simple experiment using grains (beans, corn and chickpeas) to explore different types of calibration methods.

O presente artigo retoma a noção de neutralidade proveniente da técnica clínica psicanalítica, confrontando-a com noções próximas, tais como a neutralidade científica e o neutro na semiologia de Barthes. Procuramos demonstrar como tal noção descreve bem a postura analítica, exigida tanto do paciente (associação livre) quanto do analista (atenção flutuante), e como ela nasce na prática clínica freudiana, respondendo a problemas decorrentes dela. Assim, abordaremos os temas da hipnose e da sugestão, como também aqueles ligados à contra-transferência. Defenderemos, ainda, que a neutralidade não é apenas questão de técnica, mas é central para o entendimento da própria ética da psicanálise.

O artigo discute as relações entre a psicanálise e a ciência, especialmente no que tange aos seus métodos de investigação e trabalho. Para isso, tomamos como objeto a noção de neutralidade, tal como podemos apreendê-la na clínica da psicanálise, para confrontá-la com o que chamamos de isenção no campo da ciência, que não quer dizer outra coisa que a neutralidade proposta lá. Veremos que, apesar da clara filiação da noção psicanalítica para com a noção vinda da ciência, a neutralidade clínica da psicanálise toma outro rumo, que se pode mesmo opor ao sentido do termo na ciência. Enquanto em ciência o termo quer significar um processo de purificação e de seleção, em psicanálise ele surge como a exigência de nada selecionar.

The early and correct diagnosis of human leishmaniasis is essential for disease treatment. Another important step in the control of visceral leishmaniasis is the identification of infected dogs, which are the main domestic reservoir of L. infantum. Recombinant proteins and synthetic peptides based on Leishmania genes have emerged as valuable targets for serodiagnosis due to their increased sensitivity, specificity and potential for standardization. Cathepsin L-like genes are surface antigens that are secreted by amastigotes and have little similarity to host proteins, factors that enable this protein as a good target for serodiagnosis of the leishmaniasis.We mapped a linear B-cell epitope within the Cathepsin L-like protein from L. braziliensis. A synthetic peptide containing the epitope and the recombinant protein was evaluated for serodiagnosis of human tegumentary and visceral leishmaniasis, as well as canine visceral leishmaniasis.The recombinant protein performed best for human tegumentary and canine visceral leishmaniasis, with 96.30% and 89.33% accuracy, respectively. The synthetic peptide was the best to discriminate human visceral leishmaniasis, with 97.14% specificity, 94.55% sensitivity and 96.00% accuracy. Comparison with T. cruzi-infected humans and dogs suggests that the identified epitope is specific to Leishmania parasites, which minimizes the likelihood of cross-reactions.

BACKGROUND: The present study analyzed whether or not the in vitro cultivation for long periods of time of pre-isolated Leishmania amazonensis from lesions of chronically infected BALB/c mice was able to interfere in the parasites' infectivity using in vivo and in vitro experiments. In addition, the proteins that presented a significant decrease or increase in their protein expression content were identified applying a proteomic approach. METHODOLOGY/PRINCIPAL FINDINGS: Parasites were cultured in vitro for 150 days. Aliquots were collected on the day 0 of culture (R0), as well as after ten (R10; 50 days of culture), twenty (R20; 100 days of culture), and thirty (R30; 150 days of culture) passages, and were used to analyze the parasites' in vitro and in vivo infectivity, as well as to perform the proteomic approach. Approximately 837, 967, 935, and 872 spots were found in 2-DE gels prepared from R0, R10, R20, and R30 samples, respectively. A total of 37 spots presented a significant decrease in their intensity of expression, whereas a significant increase in protein content during cultivation could be observed for 19 proteins (both cases >2.0 folds). Some of these identified proteins can be described, such as diagnosis and/or vaccine candidates, while others are involved in the infectivity of Leishmania. It is interesting to note that six proteins, considered hypothetical in Leishmania, showed a significant decrease in their expression and were also identified. CONCLUSIONS/SIGNIFICANCE: The present study contributes to the understanding that the cultivation of parasites over long periods of time may well be related to the possible loss of infectivity of L. amazonensis. The identified proteins that presented a significant decrease in their expression during cultivation, including the hypothetical, may also be related to this loss of parasites' infectivity, and applied in future studies, including vaccine candidates and/or immunotherapeutic targets against leishmaniasis.

The search toward the establishment of novel serological tests for the diagnosis of leishmaniasis and proper differential diagnosis may represent one alternative to the invasive parasitological methods currently used to identify infected individuals. In the present work, we investigated the potential use of recombinant peroxidoxin (rPeroxidoxin) of Leishmania (Viannia) braziliensis as a potential antigen for the immunodiagnosis of human tegumentary (TL) and visceral leishmaniasis (VL) and canine visceral leishmaniasis (CVL). Linear B-cell epitope mapping was performed to identify polymorphic epitopes when comparing orthologous sequences present in Trypanosoma cruzi, the agent for Chagas disease (CD), and the Homo sapiens and Canis familiaris hosts. The serological assay (ELISA) demonstrated that TL, VL and CVL individuals showed high levels of antibodies against rPeroxidoxin, allowing identification of infected ones with considerable sensitivity and great ability to discriminate (specificity) between non-infected and CD individuals (98.46% and 100%; 98.18% and 95.71%; 95.79% and 100%, respectively). An rPeroxidoxin ELISA also showed a greater ability to discriminate between vaccinated and infected animals, which is an important requirement for the public campaign control of CVL. A depletion ELISA assay using soluble peptides of this B-cell epitope confirmed the recognition of these sites only by Leishmania-infected individuals. Moreover, this work identifies two antigenic polymorphic linear B-cell epitopes of L. braziliensis. Specific recognition of TL and VL patients was confirmed by significantly decreased IgG reactivity against rPeroxidoxin after depletion of peptide-1- and peptide-2-specific antibodies (peptide 1: reduced by 32%, 42% and 5% for CL, ML and VL, respectively; peptide-2: reduced by 24%, 22% and 13% for CL, ML and VL, respectively) and only peptide-2 for CVL (reduced 9%). Overall, rPeroxidoxin may be a potential antigen for the immunodiagnosis of TL, VL or CVL, as it has a higher agreement with parasitological assays and is better than other reference tests that use soluble Leishmania antigens for diagnosing CVL in Brazil (EIE-LVC, Bio-manguinhos, FIOCRUZ).

BACKGROUND: The development of cost-effective prophylactic strategies to prevent leishmaniasis has become a high-priority. The present study has used the phage display technology to identify new immunogens, which were evaluated as vaccines in the murine model of visceral leishmaniasis (VL). Epitope-based immunogens, represented by phage-fused peptides that mimic Leishmania infantum antigens, were selected according to their affinity to antibodies from asymptomatic and symptomatic VL dogs' sera. METHODOLOGY/MAIN FINDINGS: Twenty phage clones were selected after three selection cycles, and were evaluated by means of in vitro assays of the immune stimulation of spleen cells derived from naive and chronically infected with L. infantum BALB/c mice. Clones that were able to induce specific Th1 immune response, represented by high levels of IFN-γ and low levels of IL-4 were selected, and based on their selectivity and specificity, two clones, namely B10 and C01, were further employed in the vaccination protocols. BALB/c mice vaccinated with clones plus saponin showed both a high and specific production of IFN-γ, IL-12, and GM-CSF after in vitro stimulation with individual clones or L. infantum extracts. Additionally, these animals, when compared to control groups (saline, saponin, wild-type phage plus saponin, or non-relevant phage clone plus saponin), showed significant reductions in the parasite burden in the liver, spleen, bone marrow, and paws' draining lymph nodes. Protection was associated with an IL-12-dependent production of IFN-γ, mainly by CD8+ T cells, against parasite proteins. These animals also presented decreased parasite-mediated IL-4 and IL-10 responses, and increased levels of parasite-specific IgG2a antibodies. CONCLUSIONS/SIGNIFICANCE: This study describes two phage clones that mimic L. infantum antigens, which were directly used as immunogens in vaccines and presented Th1-type immune responses, and that significantly reduced the parasite burden. This is the first study that describes phage-displayed peptides as successful immunogens in vaccine formulations against VL.

BackgroundWe evaluated kDNA PCR/hybridization and quantitative real-time PCR (qPCR) targeting the gene of DNA polymerase of Leishmania infantum for CVL diagnosis and assessment of parasite load in clinical samples obtained invasively and non-invasively.Methodology/principal findingsEighty naturally infected dogs from an endemic urban area in Brazil were used. Animals were divided into two groups based on the presence or absence of CVL clinical sings. Skin biopsies, bone marrow, blood and conjunctival swabs samples were collected and submitted to L. infantum DNA detection. In addition, anti-Leishmania antibody titers were measured by Immunofluorescence antibody test. The symptomatic dogs had increased titers compared to asymptomatic dogs (P = 0.025). The frequencies of positive results obtained by kDNA PCR/hybridization for asymptomatic and symptomatic dogs, respectively, were as follows: right conjunctiva, 77.5% and 95.0%; left conjunctiva, 75.0% and 87.5%; skin, 45.0% and 75.0%; bone marrow, 50.0% and 77.5%; and blood, 27.5% and 22.5%. In both groups, the parasite load in the skin samples was the highest (PConclusionsThe conjunctival swab is a suitable clinical sample for qualitative molecular diagnosis of CVL. The highest parasite burdens were detected in skin regardless of the presence of VL-associated clinical signs. The qPCR results emphasized the role of dogs, particularly asymptomatic dogs, as reservoirs for CVL because of the high cutaneous parasite loads. These results may help to explain the maintenance of high transmission rates and numbers of CVL cases in endemic urban regions.

BACKGROUND: The immune response in the skin of dogs infected with Leishmania infantum is poorly understood, and limited studies have described the immunopathological profile with regard to distinct levels of tissue parasitism and the clinical progression of canine visceral leishmaniasis (CVL). METHODOLOGY/PRINCIPAL FINDINGS: A detailed analysis of inflammatory cells (neutrophils, eosinophils, mast cells, lymphocytes, and macrophages) as well as the expression of chemokines (CCL2, CCL4, CCL5, CCL13, CCL17, CCL21, CCL24, and CXCL8) was carried out in dermis skin samples from 35 dogs that were naturally infected with L. infantum. The analysis was based on real-time polymerase chain reaction (PCR) in the context of skin parasitism and the clinical status of CVL. We demonstrated increased inflammatory infiltrate composed mainly of mononuclear cells in the skin of animals with severe forms of CVL and high parasite density. Analysis of the inflammatory cell profile of the skin revealed an increase in the number of macrophages and reductions in lymphocytes, eosinophils, and mast cells that correlated with clinical progression of the disease. Additionally, enhanced parasite density was correlated with an increase in macrophages and decreases in eosinophils and mast cells. The chemokine mRNA expression demonstrated that enhanced parasite density was positively correlated with the expression of CCL2, CCL4, CCL5, CCL21, and CXCL8. In contrast, there was a negative correlation between parasite density and CCL24 expression. CONCLUSIONS/SIGNIFICANCE: These findings represent an advance in the knowledge about skin inflammatory infiltrates in CVL and the systemic consequences. Additionally, the findings may contribute to the design of new and more efficient prophylactic tools and immunological therapies against CVL.

Anatomy is the branch discipline focused on studying organisms' physical structures and parts. Although technological advances are broadening the anatomy study, the practices of prosection and dissection of human cadavers and animals remain a primary teaching method. Despite the large body of research supporting its benefits, in some countries, cadaveric prosection and dissection of vertebrate animals in secondary education have been banned. In the current study, to prevent a lack of access to anatomical sciences education, the use of plastinated biological specimens was proposed for teaching practical biology in middle and high schools. The study was conducted in the 2014 academic year. Eighty-seven middle school students participated in the experiment. Groups consisted of: (i) theoretical classes only; (ii) theoretical class plus prosection with fresh specimens class; (iii) theoretical class plus expository with plastinated specimens classes. A post-test grade method was used to assess the impact of such tasks on the learning experience of each group. An ANOVA test and multiple regression model were used to analyze the effects of the variables of interest. Our study highlighted that students who underwent the plastination practical class had higher overall performance and a higher mean post-test grade than those in the pure theoretical group. A favorable effect of a positive self-knowledge assessment on the students' performance was found, supporting the self-efficacy model of human behavior. Thus, the current study provides further evidence to support the use of plastinated specimens as an effective teaching method in countries where dissection is not feasible.

Este trabalho teve por objetivo avaliar a quantidade de água que percola um NEOSSOLO FLÚVICO irrigado sob manejo racional e seu efeito na qualidade da água subterrânea. Para tanto, foi instalada uma estação de monitoramento constituída de um tubo de acesso para sonda de neutrons e um poço de observação da potenciometria do aquífero livre. A evapotranspiração foi estimada através de Tanque Classe A. O balanço foi usado para estimar a lâmina drenada para o aquífero. A irrigação promoveu a lavagem do solo e ao final do cultivo, a lâmina drenada acumulada foi cerca de 110 mm. A despeito da irrigação a potenciometria caiu durante o período estudado, com exceção dos 10 dias seguintes a precipitação de meados de outubro, onde recebeu aportes de áreas circunvizinhas. A condutividade elétrica da água subterrânea apresentou tendência de aumento durante o período estudado, indicando que a agricultura irrigada está poluindo o aquífero.

Abstract: Visceral leishmaniasis (VL) is one of the most important tropical diseases worldwide. Although chemotherapy has been widely used to treat this disease, problems related to the development of parasite resistance and side effects associated with the compounds used have been noted. Hence, alternative approaches for VL control are desirable. Some methods, such as vector control and culling of infected dogs, are insufficiently effective, with the latter not ethically recommended. The development of vaccines to prevent VL is a feasible and desirable measure for disease control; for example, some vaccines designed to protect dogs against VL have recently been brought to market. These vaccines are based on the combination of parasite fractions or recombinant proteins with adjuvants that are able to induce cellular immune responses; however, their partial efficacy and the absence of a vaccine to protect against human leishmaniasis underline the need for characterization of new vaccine candidates. This review presents recent advances in control measures for VL based on vaccine development, describing extensively studied antigens, as well as new antigenic proteins recently identified using immuno-proteomic techniques.