ABSTRACT
The objective of this study was to evaluate the water status, photosynthetic pigments, and photochemical efficiency of mini watermelon plants under salt stress and phosphate fertilization. The experiment was conducted in pots under greenhouse conditions in Pombal, PB, Brazil. The experimental design used was randomized blocks in a 5 × 4 factorial scheme, with five levels of electrical conductivity of irrigation water - ECw (0.3, 1.3, 2.3, 3.3, and 4.3 dS m-1) and four doses of phosphorus (60, 80, 100, and 120% of the recommendation), with three replicates. The relative water content in the tissues decreased with the increase in ECw levels in all phosphorus doses, with decreases of 7.05, 7.81 and 8.83% per unit increase in ECw, in plants fertilized with 80, 100 and 120% P2O5. On the other hand, ECw levels increased electrolyte leakage, regardless of phosphorus doses of the recommendation. The synthesis of photosynthetic pigments and the quantum efficiency of photosystem II were inhibited by increasing water salinity from 0.3 dS m-1 in plants grown under phosphorus doses above 60% of the recommendation. Water salinity from 0.3 dS m-1 reduced chlorophyll b contents, initial, maximum, and variable fluorescence of mini watermelon plants, with a decrease of 11.86, 4.51, 4.53, and 4.54% per unit increment of ECw, respectively.
Subject(s)
Citrullus , Phosphates , Photosynthesis , Saline Waters , PhosphorusABSTRACT
The limitation in the quality of water sources for irrigation in the semi-arid region of northeastern Brazil is increasingly present, so it is necessary to use water with high concentrations of salts for agricultural production, which makes the use of elicitors essential to mitigate the harmful effects of salinity on plants. Given the above, the objective of this study was to evaluate the effects of foliar application of salicylic acid on the mineral composition and production of guava plants under salt stress conditions in the post-grafting phase. The experiment was carried out under greenhouse conditions, in a randomized block design, in a 2 × 4 factorial scheme, with two levels of electrical conductivity of irrigation water (0.6 and 3.2 dS m-1) and four concentrations of salicylic acid (0, 1.2, 2.4, and 3.6 mM), with three replicates. During the flowering stage of guava, N, P, and K contents accumulated in the leaves according to the following order of concentration: N > K > P. Foliar application of 1.2 mM of salicylic acid increases the leaf contents of N, P, and K in guava plants grown under irrigation with water of 0.6 dS m-1. Water salinity of 3.2 dS m-1 reduces the growth and production components of guava plants.
Subject(s)
Psidium , Salicylic Acid , Salt Stress , Minerals , WaterABSTRACT
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Subject(s)
Trematoda , Trematode Infections , Animals , Trematode Infections/veterinary , Trematode Infections/parasitology , Microscopy, Electron, Scanning , Columbidae/parasitology , KidneyABSTRACT
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).
Subject(s)
Animals , Trematoda/anatomy & histology , Body Weights and Measures/veterinary , Microscopy, Electron, ScanningABSTRACT
The limitation in the quality of water sources for irrigation in the semi-arid region of northeastern Brazil is increasingly present, so it is necessary to use water with high concentrations of salts for agricultural production, which makes the use of elicitors essential to mitigate the harmful effects of salinity on plants. Given the above, the objective of this study was to evaluate the effects of foliar application of salicylic acid on the mineral composition and production of guava plants under salt stress conditions in the post-grafting phase. The experiment was carried out under greenhouse conditions, in a randomized block design, in a 2 × 4 factorial scheme, with two levels of electrical conductivity of irrigation water (0.6 and 3.2 dS m-1) and four concentrations of salicylic acid (0, 1.2, 2.4, and 3.6 mM), with three replicates. During the flowering stage of guava, N, P, and K contents accumulated in the leaves according to the following order of concentration: N > K > P. Foliar application of 1.2 mM of salicylic acid increases the leaf contents of N, P, and K in guava plants grown under irrigation with water of 0.6 dS m-1. Water salinity of 3.2 dS m-1 reduces the growth and production components of guava plants.
A limitação na qualidade das fontes hídricas para irrigação na região semiárida do Nordeste é cada vez mais presente, tornando-se necessário o uso de águas com altas concentrações de sais para produção agrícola, o que torna indispensável à utilização de elicitores para atenuar os efeitos deletérios da salinidade sobre as plantas. Diante do exposto, objetivou-se com esse trabalho avaliar os efeitos da aplicação foliar de ácido salicílico na composição mineral e na produção de plantas de goiabeira sob condições de estresse salino na fase pós-enxertia. O experimento foi conduzido em casa de vegetação, no delineamento experimental de blocos casualizados, em esquema fatorial 2 × 4, sendo dois níveis de condutividade elétrica da água de irrigação (0,6 e 3,2 dS m-1) e quatro concentrações de ácido salicílico (0; 1,2; 2,4 e 3,6 mM), com três repetições. Durante a fase de floração da goiabeira, os teores de N, P, K, se acumularam nas folhas segundo a ordem de concentração N > K > P. A aplicação foliar de 1,2 mM de ácido salicílico aumentou os teores foliares de nitrogênio, fósforo e potássio nas plantas de goiabeira cultivadas sob irrigação com água de 0,6 dS m-1. A salinidade da água de 3,2 dS m-1 diminuiu o crescimento e os componentes de produção das plantas de goiabeira.
Subject(s)
Salicylic Acid , Psidium , Salt Stress , MineralsABSTRACT
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).
Subject(s)
Animals , Columbidae/parasitology , Trematoda/anatomy & histology , Microscopy, Electron, ScanningABSTRACT
Trypanosoma cruzi proliferative forms perform endocytosis through a specialized structure named the cytostome-cytopharynx complex (SPC). The SPC is a specialized invagination of the cell membrane that extends through the cell body towards the posterior regions, with its aperture close to the flagellar pocket. Recently, diverse proteins were found along the cytopharynx, including two myosin motors. One of these is the orphan myosin MyoF, that was proved to be essential for endocytosis in epimastigotes. However, the dynamics of MyoF localization along the endocytic pathway and through the T. cruzi life cycle remain unclear. Using CRISPR-Cas9 genome editing, we generated epimastigotes expressing MyoF fused to mNeonGreen from its endogenous locus. Using these cells, we observed that during the epimastigote cell cycle MyoF signal disappeared during G2, reappearing at early cytokinesis. Additionally, we show that MyoF localization during metacyclogenesis is compatible with the progressive disappearance of the SPC, being absent in metacyclic trypomastigotes. Detergent fractionation showed that MyoF was predominantly present in the insoluble fraction and immunolocalized at the SPC microtubules in whole-mount cytoskeleton preparations. Moreover, during tracer uptake through the SPC, MyoF followed the tracer along the endocytic pathway and was found in posterior compartments after 30 min. Taken together, the data suggest that MyoF may play a role not only at the cargo entry site but also along the endocytic pathway.
Subject(s)
Endocytosis , Myosins/genetics , Protozoan Proteins/genetics , Trypanosoma cruzi/physiology , Myosins/metabolism , Protozoan Proteins/metabolismABSTRACT
O tamanduá-bandeira (Myrmecophaga tridactyla) e o tamanduá-mirim (Tamandua tetradactyla) são mamíferos xenartros amplamente distribuídos no território nacional. Apesar disso, suas populações estão em constante ameaça, principalmente o primeiro, categorizado como vulnerável de acordo com classificação da União Internacional para Conservação da Natureza. Diante disso e da escassez de estudos com endoparasitas nesses animais, objetivou-se descrever a helmintofauna dessas duas espécies de tamanduá, utilizando espécimes atropelados nas rodovias BR-050 e BR-455, região do Triângulo Mineiro, Minas Gerais, Brasil. Os animais foram submetidos à necropsia, tendo seus órgãos e conteúdo intestinal e estomacal examinados para recuperação de helmintos. Os parasitos coletados foram identificados com base em suas características morfológicas e morfométricas. As espécies/os gêneros caracterizados para M. tridactyla e T. tetradactyla, suas respectivas prevalências, bem como órgãos de origem, foram: Gaphidiops dissimilis (33,33% e 0%) no estômago; Gigantorhynchus echinodiscus (25% e 66,66%) no intestino delgado; Physaloptera magnipapilla (50% e 66,6%) no estômago; e Mathevotaenia spp. (25% e 66,66%) no intestino delgado. Physaloptera magnipapilla foi a espécie com os maiores índices de infecção, e novo registro de localização foi registrado para o nematódeo Graphidiops dissimilis. Detectou-se uma provável sobreposição da área de vida dos tamanduás estudados, bem como uma helmintofauna característica da região.(AU)
Myrmecophaga tridactyla, giant anteater and Tamandua tetradactyla, collared anteater are nationally widespread mammals. Despite that, their populations are in danger, especially the first one, categorized as vulnerable according to the International Union for Conservation of Nature. This study reports the occurrence of helminths in these two anteater species using road killed specimens from BR-050 and BR-455 highways, Triângulo Mineiro region, Minas Gerais state, Brazil. The species were necropsied, and their gastrointestinal organs and contents were opened and examined for parasite recovery. The identification was based on morphological and morphometric features. The species/ genera found for M. tridactyla and T. tetradactyla, their prevalence, as well the organs, were: Gaphidiops dissimilis (33.33% and 0%) in stomach; Gigantorhynchus echinodiscus (25% and 66.66%) in small intestine; Physaloptera magnipapilla (50% and 66.6%) in stomach and Mathevotaenia spp. (25% and 66.66%) in small intestine. Physaloptera magnipapilla was the most common species and a new location was reported for Graphidiops dissimilis. There is a probable overlap in the home range of these animals and a local helminth diversity was described on the study site.(AU)
Subject(s)
Animals , Spirurina , Eutheria/parasitology , Helminths/isolation & purification , Nematoda/isolation & purification , Autopsy/veterinary , BrazilABSTRACT
OBJECTIVES: This study aimed to identify, describe and analyze priority areas for COVID-19 testing combining participatory surveillance and traditional surveillance. DESIGN: It was carried out a descriptive transversal study in the city of Caruaru, Pernambuco state, Brazil, within the period of 20/02/2020 to 05/05/2020. Data included all official reports for influenza-like illness notified by the municipality health department and the self-reports collected through the participatory surveillance platform Brasil Sem Corona. METHODS: We used linear regression and loess regression to verify a correlation between Participatory Surveillance (PS) and Traditional Surveillance (TS). Also a spatial scanning approach was deployed in order to identify risk clusters for COVID-19. RESULTS: In Caruaru, the PS had 861 active users, presenting an average of 1.2 reports per user per week. The platform Brasil Sem Corona started on March 20th and since then, has been officially used by the Caruaru health authority to improve the quality of information from the traditional surveillance system. Regarding the respiratory syndrome cases from TS, 1588 individuals were positive for this clinical outcome. The spatial scanning analysis detected 18 clusters and 6 of them presented statistical significance (p-value < 0.1). Clusters 3 and 4 presented an overlapping area that was chosen by the local authority to deploy the COVID-19 serology, where 50 individuals were tested. From there, 32 % (nâ¯=â¯16) presented reagent results for antibodies related to COVID-19. CONCLUSION: Participatory surveillance is an effective epidemiological method to complement the traditional surveillance system in response to the COVID-19 pandemic by adding real-time spatial data to detect priority areas for COVID-19 testing.
Subject(s)
COVID-19 Testing , COVID-19/diagnosis , Coronavirus Infections/diagnosis , Population Surveillance , Adolescent , Adult , Algorithms , Brazil/epidemiology , Humans , Linear Models , Middle Aged , Pandemics , SARS-CoV-2 , Self Report , Spatial Analysis , Young AdultABSTRACT
INTRODUCTION: Clonogenic assay evaluates the potential of cells to undergo division or generate clones following treatment with a chemical or other agent, thereby allowing the evaluation of cytotoxic and/or antiproliferative effects. Clonogenic assay analysis using traditional methods tends to be time-consuming and yield inconsistent results, whereas results from analyses conducted using automated image processing methods may be misleading or subject to misinterpretation. Thus, the aim of this work was to validate and demonstrate the applicability of a recently developed software. METHODS: Repeatability of measurements was evaluated by comparing results from 10 replicate images from a single well. To evaluate the viability of the software, results were compared with those obtained from manual counting, crystal violet optical density, and up-to-date automated methods. A clonogenic index was experimentally developed using the individual area occupied by colonies, while clone stratification was used to differentiate between antiproliferative and cytotoxic effects. RESULTS: The developed software showed to be a reliable and consistent tool for clonogenic assay evaluation, presenting a repeatability mean error of 0.79% for the number of colonies and 0.89% for the total area of colonies, as well as exhibiting a significant correlation (p < 0.05) with results obtained from widely adopted gold standard methods. The software was also able to detect an appropriate dose-dependent effect as well as a predominant cytotoxic effect of vincristine on MCF-7 cells and calculate the clonogenic index. DISCUSSION: Therefore, this software is adequate for the analysis of clonogenic assay images, differentiating between cytotoxic and antiproliferative trends.
Subject(s)
Image Processing, Computer-Assisted/methods , Intravital Microscopy/methods , Software , Tumor Stem Cell Assay/methods , Antineoplastic Agents, Phytogenic/pharmacology , Cell Count/methods , Cell Proliferation/drug effects , Drug Screening Assays, Antitumor/methods , Humans , MCF-7 Cells , Reproducibility of Results , Vincristine/pharmacologyABSTRACT
Caddisflies are a highly diverse group of aquatic insects, particularly in the Neotropical region where there is a high number of endemic taxa. Based on taxonomic contributions published until August 2019, a total of 796 caddisfly species have been recorded from Brazil. Taxonomic data about Brazilian caddisflies are currently open access at the Catálogo Taxonômico da Fauna do Brasil website (CTFB), an on-line database with taxonomic information on the animal species occurring in Brazil. The order Trichoptera at CTFB includes a catalog of species recorded for the country, with synonymic lists, distribution throughout six biomes, 12 hydrographic regions, and 27 political states (including Federal District) from Brazil. The database is constantly updated to include newly published data. In this study, we reviewed the taxonomic effort on Brazilian caddisflies based on data currently in CTFB database. The accumulation curve of species described or recorded from the country, by year, shows a strong upward trend in last 25 years, indicating that it is possible that there are many more species to be described. Based on presence/absence of caddisfly species at three geographic levels (biomes, hydrographic regions, and states), second order Jackknife estimated at least 1,586 species occurring in Brazil (with hydrographic regions as unities), indicating we currently know about 50% of the Brazilian caddisfly fauna. Species distribution by Brazilian biomes reveals that the Atlantic Forest is the most diverse, with 490 species (298 endemic), followed by the Amazon Forest, with 255 species (101 endemic). Even though these numbers may be biased because there has been more intense collecting in these two biomes, the percentage of endemic caddisfly species in the Atlantic Forest is remarkable. Considering the distribution throughout hydrographic regions, clustering analyses (UPGMA) based on incidence data reveals two groups: northwestern basins and southeastern. Although these groups have weak bootstrap support and low similarity in species composition, this division of Brazilian caddisfly fauna could be related to Amazon-Atlantic Forest disjunction, with the South American dry diagonal acting as a potential barrier throughout evolutionary time.
Subject(s)
Animals , Biodiversity , Aquatic Fauna , InsectaABSTRACT
Caddisflies are a highly diverse group of aquatic insects, particularly in the Neotropical region where there is a high number of endemic taxa. Based on taxonomic contributions published until August 2019, a total of 796 caddisfly species have been recorded from Brazil. Taxonomic data about Brazilian caddisflies are currently open access at the Catálogo Taxonômico da Fauna do Brasil website (CTFB), an on-line database with taxonomic information on the animal species occurring in Brazil. The order Trichoptera at CTFB includes a catalog of species recorded for the country, with synonymic lists, distribution throughout six biomes, 12 hydrographic regions, and 27 political states (including Federal District) from Brazil. The database is constantly updated to include newly published data. In this study, we reviewed the taxonomic effort on Brazilian caddisflies based on data currently in CTFB database. The accumulation curve of species described or recorded from the country, by year, shows a strong upward trend in last 25 years, indicating that it is possible that there are many more species to be described. Based on presence/absence of caddisfly species at three geographic levels (biomes, hydrographic regions, and states), second order Jackknife estimated at least 1,586 species occurring in Brazil (with hydrographic regions as unities), indicating we currently know about 50% of the Brazilian caddisfly fauna. Species distribution by Brazilian biomes reveals that the Atlantic Forest is the most diverse, with 490 species (298 endemic), followed by the Amazon Forest, with 255 species (101 endemic). Even though these numbers may be biased because there has been more intense collecting in these two biomes, the percentage of endemic caddisfly species in the Atlantic Forest is remarkable. Considering the distribution throughout hydrographic regions, clustering analyses (UPGMA) based on incidence data reveals two groups: northwestern basins and southeastern. Although these groups have weak bootstrap support and low similarity in species composition, this division of Brazilian caddisfly fauna could be related to Amazon-Atlantic Forest disjunction, with the South American dry diagonal acting as a potential barrier throughout evolutionary time.(AU)
Subject(s)
Animals , Insecta , Biodiversity , Aquatic FaunaABSTRACT
Toxoplasma gondii, the agent of toxoplasmosis, is an intracellular parasite that can infect a wide range of vertebrate hosts. Toxoplasmosis causes severe damage to immunocompromised hosts and its treatment is mainly based on the combination of pyrimethamine and sulfadiazine, which causes relevant side effects primarily observed in AIDS patients, including bone marrow suppression and hematological toxicity (pyrimethamine) and/or hypersensitivity and allergic skin reactions (sulfadiazine). Thus, it is important to investigate new compounds against T. gondii, particularly those that may act on bradyzoites, which are present in cysts during the chronic disease phase. We propose an in vitro model to simultaneously study new candidate compounds against the two main causative stages of Toxoplasma infection in humans, using the EGS-DC strain that was modified from a type I/III strain (EGS), isolated from a case of human congenital toxoplasmosis in Brazil and engineered to express markers for both stages of development. One feature of this strain is that it presents tachyzoite and bradyzoite in the same culture system and in the same host cell under normal culture conditions. Additionally, this strain presents stage-specific fluorescent protein expression, allowing for easy identification of both stages, thus making this strain useful in different studies. HFF cells were infected and after 4 and 7 days post infection the cells were treated with 10 µM of pyrimethamine or atovaquone, for 48 or 72 h. We used high-throughput screening to quantify the extent of parasite infection. Despite a reduction in tachyzoite infection caused by both treatments, the atovaquone treatment reduced the bradyzoite infection while the pyrimethamine one increased it. Ultrastructural analysis showed that after treatment with both drugs, parasites displayed altered mitochondria. Fluorescence microscopy of cells labeled with MitoTracker CMXRos showed that the cysts present inside the cells lost their mitochondrial membrane potential. Our results indicate that this experimental model is adequate to simultaneously analyze new active compounds against tachyzoite and bradyzoite forms.
Subject(s)
Parasitology/methods , Toxoplasma/growth & development , Toxoplasma/genetics , Toxoplasmosis, Congenital/parasitology , Antiprotozoal Agents/pharmacology , Atovaquone/pharmacology , Brazil , Cell Line , Genetic Markers , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Humans , Life Cycle Stages , Pyrimethamine/pharmacology , Toxoplasma/drug effects , Toxoplasma/metabolism , Toxoplasmosis, Congenital/diagnosisABSTRACT
Titanium (Ti) and its alloys are widely used in dental implants and hip-prostheses due to their excellent biocompatibility. Growing evidence support that surface degradation due to corrosion and wear processes, contribute to implant failure, since the release of metallic ions and wear particles generate local tissue reactions (peri-implant inflammatory reactions). The generated ions and wear debris (particles at the micron and nanoscale) stay, in a first moment, at the interface implant-bone. However, depending on their size, they can enter blood circulation possibly contributing to systemic reactions and toxicities. Most of the nanotoxicological studies with titanium dioxide nanoparticles (TiO2 NPs) use conventional two-dimensional cell culture monolayers to explore macrophage and monocyte activation, where limited information regarding bone cells is available. Recently three-dimensional models have been gaining prominence since they present a greater anatomical and physiological relevance. Taking this into consideration, in this work we developed a human osteoblast-like spheroid model, which closely mimics bone cell-cell interactions, providing a more realistic scenario for nanotoxicological studies. The treatment of spheroids with different concentrations of TiO2 NPs during 72 h did not change their viability significantly. Though, higher concentrations of TiO2 NPs influenced osteoblast cell cycle without interfering in their ability to differentiate and mineralize. For higher concentration of TiO2 NPs, collagen deposition and pro-inflammatory cytokine, chemokine and growth factor secretion (involved in osteolysis and bone homeostasis) increased. These results raise the possible use of this model in nanotoxicological studies of osseointegrated devices and demonstrate a possible therapeutic potential of this TiO2 NPs to prevent or reverse bone resorption.
Subject(s)
Nanoparticles/toxicity , Osteoblasts/cytology , Spheroids, Cellular/cytology , Spheroids, Cellular/drug effects , Titanium/pharmacology , Titanium/toxicity , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Line , Cell Survival/drug effects , Cytokines/metabolism , Dose-Response Relationship, Drug , Homeostasis/drug effects , Humans , Minerals/metabolism , Spheroids, Cellular/metabolism , Titanium/chemistryABSTRACT
Gastrointestinal nematodes are important ecological assets for the maintenance of the biodiversity in the Atlantic Forest in Brazil. They parasitize a number of animals of the local fauna, in which some species can promote serious injuries in the stomach wall of their hosts, which may lead to death. Among these nematodes, parasites of the genus Physaloptera are known to parasitize mammals (particularly carnivores and small rodents), birds and reptiles, being important for the local biodiversity. In this work, three hundred and sixty-two nematodes were recovered from the stomach of twenty-one Metachirus nudicaudatus (Didelphimorphia: Didelphidae) collected in Duas Bocas Biological Reserve, State of Espírito Santo, one of the largest Atlantic Forest remnants and important wildlife refuge of the Atlantic Forest in Brazil. Analysis using fluorescence and scanning electron microscopy as well as phylogenetic assessment using the mitochondrial cytochrome c oxidase subunit I gene showed that the parasites belong to the Physaloptera. Our results show details of the nematode morphology including the cloacal papillae distribution, cuticular topography details, 2D and 3D measurements of the structures with taxonomic importance. Molecular data confirmed the validity of P. mirandai and the phylogeny supported the monophyly of the assemblage formed by Physaloptera and Turgida. The use of a combination of quantitative and multidimensional microscopy tools, such as 3D reconstruction and modeling, allied to phylogenetic analysis may provide grounds for a new approach on helminth taxonomy and structural characterization.
Subject(s)
Anatomy, Veterinary/methods , Imaging, Three-Dimensional/methods , Microscopy, Electron, Scanning/methods , Phylogeny , Spiruroidea/classification , Spiruroidea/genetics , Animals , BrazilABSTRACT
A pesquisa teve como objetivo avaliar a qualidade da carne de tambaqui com diferentes pesos de abate. Foram utilizados vinte e quatro tambaquis, entre 1,0 e 3,5 kg. O peso ao abate não afetou o pH da carne (6,42), a cor para luminosidade (L*), intensidade de vermelho (a*) e amarelo (b*), respectivamente, com médias de 62,00; 2,86 e 15,57; e os atributos sensoriais aparência (7,41), sabor (7,25), textura (7,40) e aceitação global (7,24). A perda de peso por gotejamento e descongelamento e a capacidade de retenção de água da carne de tambaqui foram afetados (P<0,01) pelo peso ao abate. Embora o peso de abate do tambaqui entre 1,0 e 1,5 kg apresente maiores perdas de água, as demais características qualitativas da carne não foram comprometidas.
The experiment was conducted to evaluate the meat quality of tambaqui slaughtered at different weights. Twenty-four animals were slaughtered at body weights ranging from 1.0 to 3.5 kg. The slaughter weight did not affect meat pH (6.42), luminosity (L*), red color intensity (a*) or yellow color intensity (b*) (means of 62.0, 2.86 and 15.57, respectively), nor the sensory attributes appearance (7.41), flavor (7.25), texture (7.40), and overall acceptance (7.24). Weight loss by dripping and defrosting and the water holding capacity of tambaqui meat were affected (P<0.01) by slaughter weight. Although a slaughter weight of tambaqui between 1.0 and 1.5 kg was associated with further water loss, the other meat quality traits were not compromised.
Subject(s)
Animals , Meat/analysis , Body Weights and Measures , Fish Products/analysis , Food Quality , Animal Culling , Characidae , ColorABSTRACT
A pesquisa teve como objetivo avaliar a qualidade da carne de tambaqui com diferentes pesos de abate. Foram utilizados vinte e quatro tambaquis, entre 1,0 e 3,5 kg. O peso ao abate não afetou o pH da carne (6,42), a cor para luminosidade (L*), intensidade de vermelho (a*) e amarelo (b*), respectivamente, com médias de 62,00; 2,86 e 15,57; e os atributos sensoriais aparência (7,41), sabor (7,25), textura (7,40) e aceitação global (7,24). A perda de peso por gotejamento e descongelamento e a capacidade de retenção de água da carne de tambaqui foram afetados (P<0,01) pelo peso ao abate. Embora o peso de abate do tambaqui entre 1,0 e 1,5 kg apresente maiores perdas de água, as demais características qualitativas da carne não foram comprometidas.(AU)
The experiment was conducted to evaluate the meat quality of tambaqui slaughtered at different weights. Twenty-four animals were slaughtered at body weights ranging from 1.0 to 3.5 kg. The slaughter weight did not affect meat pH (6.42), luminosity (L*), red color intensity (a*) or yellow color intensity (b*) (means of 62.0, 2.86 and 15.57, respectively), nor the sensory attributes appearance (7.41), flavor (7.25), texture (7.40), and overall acceptance (7.24). Weight loss by dripping and defrosting and the water holding capacity of tambaqui meat were affected (P<0.01) by slaughter weight. Although a slaughter weight of tambaqui between 1.0 and 1.5 kg was associated with further water loss, the other meat quality traits were not compromised.(AU)
Subject(s)
Animals , Food Quality , Fish Products/analysis , Meat/analysis , Body Weights and Measures , Characidae , Color , Animal CullingSubject(s)
Hematology/education , Biomedical Research , Brazil , Capacity Building , Humans , Internship and Residency , PovertyABSTRACT
Toxoplasma gondii is an intracellular protozoan parasite that causes toxoplasmosis, a prevalent infection related to abortion, ocular diseases and encephalitis in immuno-compromised individuals. In the untreatable (and life-long) chronic stage of toxoplasmosis, parasitophorous vacuoles (PVs, containing T. gondii tachyzoites) transform into tissue cysts, containing slow-dividing bradyzoite forms. While acute-stage infection with tachyzoites involves global rearrangement of the host cell cytoplasm, focused on favouring tachyzoite replication, the cytoplasmic architecture of cells infected with cysts had not been described. Here, we characterized (by fluorescence and electron microscopy) the redistribution of host cell structures around T. gondii cysts, using a T. gondii strain (EGS) with high rates of spontaneous cystogenesis in vitro. Microtubules and intermediate filaments (but not actin microfilaments) formed a 'cage' around the cyst, and treatment with taxol (to inhibit microtubule dynamics) favoured cystogenesis. Mitochondria, which appeared adhered to the PV membrane, were less closely associated with the cyst wall. Endoplasmic reticulum (ER) profiles were intimately associated with folds in the cyst wall membrane. However, the Golgi complex was not preferentially localized relative to the cyst, and treatment with tunicamycin or brefeldin A (to disrupt Golgi or ER function, respectively) had no significant effect on cystogenesis. Lysosomes accumulated around cysts, while early and late endosomes were more evenly distributed in the cytoplasm. The endocytosis tracer HRP (but not BSA or transferrin) reached bradyzoites after uptake by infected host cells. These results suggest that T. gondii cysts reorganize the host cell cytoplasm, which may fulfil specific requirements of the chronic stage of infection.