Artigos - Parasitologia - IBB

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Bacterial Microbiota from Lab-Reared and Field-Captured Anopheles darlingi Midgut and Salivary Gland
(2023-05-01) Santos, Najara Akira Costa dos; Carvalho, Vanessa Rafaela de [UNESP]; Souza-Neto, Jayme A. [UNESP]; Alonso, Diego Peres [UNESP]; Ribolla, Paulo Eduardo Martins [UNESP]; Medeiros, Jansen Fernandes; Araujo, Maisa da Silva; Fundação Universidade Federal de Rondônia/Fiocruz Rondônia; Fiocruz Rondônia; Universidade Estadual Paulista (UNESP); Fundação Universidade Federal de Rondônia; Centro de Pesquisa em Medicina Tropical
Anopheles darlingi is a major malaria vector in the Amazon region and, like other vectors, harbors a community of microorganisms with which it shares a network of interactions. Here, we describe the diversity and bacterial composition from the midguts and salivary glands of lab-reared and field-captured An. darlingi using metagenome sequencing of the 16S rRNA gene. The libraries were built using the amplification of the region V3–V4 16S rRNA gene. The bacterial community from the salivary glands was more diverse and richer than the community from the midguts. However, the salivary glands and midguts only showed dissimilarities in beta diversity between lab-reared mosquitoes. Despite that, intra-variability was observed in the samples. Acinetobacter and Pseudomonas were dominant in the tissues of lab-reared mosquitoes. Sequences of Wolbachia and Asaia were both found in the tissue of lab-reared mosquitoes; however, only Asaia was found in field-captured An. darlingi, but in low abundance. This is the first report on the characterization of microbiota composition from the salivary glands of An. darlingi from lab-reared and field-captured individuals. This study can provide invaluable insights for future investigations regarding mosquito development and interaction between mosquito microbiota and Plasmodium sp.
Zinc methionine or zinc sulphate supplementation modulate the development of the hypopharyngeal gland and expression of major royal jelly protein genes in Apis mellifera L. bees
(2023-01-01) do Prado Ribeiro, Giovanna [UNESP]; Kadri, Samir Moura [UNESP]; Justulin, Luis Antônio [UNESP]; Ribolla, Paulo Eduardo Martins [UNESP]; Orsi, Ricardo de Oliveira [UNESP]; Universidade Estadual Paulista (UNESP)
We analysed whether the source of zinc (inorganic and organic) influences the morphology of the hypopharyngeal gland (HPG) and the relative expression of major royal jelly protein (MRJP) genes in nursing bees at 6 days of age. For this, 15 colonies of Apis mellifera bees were subjected to the following treatments: Zn0 (control), zinc methionine (50 ppm) and zinc sulphate monohydrate (50 ppm) during the off-season (characterized by food reduction). MRJP1 and MRJP5 gene expression was analysed using real-time PCR and HPG sections under a microscope. Bee mortality and population development were also evaluated for 30 days. It was observed that the honeybees consumed all sugar syrup containing zinc both in its organic and inorganic forms, which did not promote mortality or affect population development. Regarding HPG morphometry, it was observed that zinc, regardless of the source, promoted the acini area significantly greater than that of the control. Zn methionine promoted a significant increase in HPG area compared with that of an inorganic source. The relative expression of MRJP1 and MRJP5 was not significantly affected compared with the control. We conclude that the mineral Zn promotes an increase in the HPG area and does not alter the pattern of expression of the MRJP1 and MRJP5 genes, and that the organic source of the mineral presents the best results.
Host Feeding Patterns of Mansonia (Diptera, Culicidae) in Rural Settlements near Porto Velho, State of Rondonia, Brazil
(2023-03-01) Alonso, Diego Peres [UNESP]; Amorim, Jandui Almeida; de Oliveira, Tatiane Marques Porangaba; de Sá, Ivy Luizi Rodrigues; Possebon, Fábio Sossai [UNESP]; de Carvalho, Dario Pires; Ribeiro, Kaio Augusto Nabas; Ribolla, Paulo Eduardo Martins [UNESP]; Sallum, Maria Anice Mureb; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); Santo Antônio Energia
Mosquito females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals since they are voraciously hematophagous and feed on the blood of a variety of vertebrates. Despite their relevance, there is a lack of investigation into the blood-feeding patterns of the Mansonia species. Knowledge of the host preference is crucial in establishing the public health importance of a mosquito species and its potential to be involved in the transmission dynamics of pathogens. Species that are primarily anthropophilic can be more effective in spreading vector-borne pathogens to humans. In this study, we used an Illumina Nextera sequencing protocol and the QIIME2 workflow to assess the diversity of DNA sequences extracted in the ingested blood of mosquito species to evaluate the overall and local host choices for three species: Ma. titillans, Ma. Amazonensis, and Ma. humeralis, in rural areas alongside the Madeira River in the vicinities of the Santo Antonio Energia (SAE) reservoir in the municipality of Porto Velho, Rondônia, Western Brazil. By performing our analysis pipeline, we have found that host diversity per collection site showed a significant heterogeneity across the sample sites. In addition, in rural areas, Ma. amazonensis present a high affinity for B. taurus, Ma. humeralis shows an overall preference for C. familiaris and B. taurus, but also H. sapiens and E. caballus in urban areas, and Ma. titillans showed more opportunistic behavior in rural areas, feeding on wild animals and G. gallus, though with an overall preference for H. sapiens.
Sporothrix pathogenic clade: Molecular analysis of animal and human clinical isolates
(2023-01-01) Ribeiro-Marques, Mariela Domiciano; Maschio-Lima, Taiza [UNESP]; Lemes, Thiago Henrique [UNESP]; Siqueira, João Paulo Zen; Brizzotti-Mazuchi, Natália Seron; Caetano, Maicon Henrique [UNESP]; Almeida, Bianca Gottardo; Mozaner, Leticia Queiroz [UNESP]; Monteiro, Ruan Campos; Camargo, Zoilo Pires; Rodrigues, Anderson Messias; Gottardo De Almeida, Margarete Teresa [UNESP]; São José Do Rio Preto School of Medicine (FAMERP); Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP)
Sporotrichosis is a subcutaneous mycosis that affects animals and humans. Varying in severity, occurrences range from local lesions to systemic involvement. It is caused by thermodimorphic and saprobic fungi from the Sporothrix pathogenic clade. This study aimed to identify the species and the sexual idiomorph distribution patterns responsible for diagnosed cases of sporotrichosis in São José do Rio Preto, Brazil. We included 188 isolates of Sporothrix sp. from feline lesions and 27 of human origin, which underwent molecular identification and genotyping for mating-type MAT1-1 and MAT1-2. The results showed that Sporothrix brasiliensis is the prevalent species in feline sporotrichosis outbreaks with the overwhelming presence of a single mating-type, MAT1-2 (P <.0001), suggesting a prevalently clonal form of spread. Morphological analyses did not discriminate among cryptic species in the genus Sporothrix, and molecular identification was essential for the correct identification of the species responsible for the observed cases of sporotrichosis. Distribution analyses of MAT1-2 isolates support the hypothesis of unidirectional migration from the current epidemics in São Paulo and Rio de Janeiro to the municipality of São José do Rio Preto.
Comparative leaf anatomy of Palicourea rigida and Palicourea coriacea (Rubiaceae) occurring in the municipality of Diorama, Goiás, Brazil
(2022-01-01) da Silva Mariano, Mayara Stefany; Mendes, Thainara Policarpo [UNESP]; Universidade Federal de Goiás (UFG); Universidade Estadual Paulista (UNESP)
This study aimed to perform a comparative anatomical study between Palicourea coriacea and Palicourea rigida (Rubiaceae), aiming to correlate possible anatomical changes to the variations of climatic factors found in the biome, and contribute to the taxonomic delimitation of the genus. The material was collected according to the usual recommendations in plant anatomy in an area of Cerrado sensu stricto, located in the municipality of Diorama – Goiás. We observed that the studied species share morphoanatomical diagnostic characteristics for the Rubiaceae family. However, some variations were found, such as styloid crystals, which were only found for species of the Psychotria genus, and which were considered a distinctive feature of the Psychotria genus. As styloid crystals were found in both P. coriacea and P. rigida, we suggest that it be used as a distinguishing feature of the tribe.
Concomitant TP53 mutation in early-stage resected EGFR-mutated non-small cell lung cancer: a narrative approach in a genetically admixed Brazilian cohort
(2023-01-01) Machado-Rugolo, J. [UNESP]; Baldavira, C. M.; Prieto, T. G.; Olivieri, E. H.R.; Fabro, A. T.; Rainho, C. A. [UNESP]; Castelli, E. C. [UNESP]; Ribolla, P. E.M. [UNESP]; Ab’saber, A. M.; Takagaki, T.; Nagai, M. A.; Capelozzi, V. L.; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); AC Camargo Cancer Center; Instituto do Câncer de São Paulo
TP53 mutations are frequent in non-small cell lung cancer (NSCLC) and have been associated with poor outcome. The prognostic and predictive relevance of EGFR/TP53 co-mutations in NSCLC is controversial. We analyzed lung tissue specimens from 70 patients with NSCLC using next-generation sequencing to determine EGFR and TP53 status and the association between these status with baseline patient and tumor characteristics, adjuvant treatments, relapse, and progression-free (PFS) and overall survival (OS) after surgical resection. We found the EGFR mutation in 32.9% of patients (20% classical mutations and 12.9% uncommon mutations). TP53 missense mutations occurred in 25.7% and TP53/EGFR co-mutations occurred in 43.5% of patients. Stage after surgical resection was significantly associated with OS (P=0.028). We identified an association between progression-free survival and poor outcome in patients with distant metastases (P=0.007). We found a marginally significant difference in OS between genders (P=0.057) and between mutant and wild type TP53 (P=0.079). In univariate analysis, distant metastases (P=0.027), pathological stage (IIIA-IIIB vs I-II; P=0.028), and TP53 status (borderline significance between wild type and mutant; P=0.079) influenced OS. In multivariable analysis, a significant model for high risk of death and poor OS (P=0.029) selected patients in stage IIIA-IIIB, with relapse and distant metastases, non-responsive to platin-based chemotherapy and erlotinib, with tumors harboring EGFR uncommon mutations, with TP53 mutant, and with EGFR/TP53 co-mutations. Our study suggested that TP53 mutation tends to confer poor survival and a potentially negative predictive effect associated with a non-response to platinum-based chemotherapy and erlotinib in early-stage resected EGFR-mutated NSCLC.
Hemogregarine Diversity Infecting Brazilian Turtles with a Description of Six New Species of Haemogregarina (Apicomplexa: Adeleorina: Haemogregarinidae)
(2023-02-01) Úngari, Letícia Pereira [UNESP]; Santos, André Luiz Quagliatto; da Silva, Reinaldo José [UNESP]; O’Dwyer, Lucia Helena [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal de Uberlândia (UFU)
(1) Background: Hemoparasites of the genus Haemogregarina (Haemogregarinidae) are commonly reported in freshwater turtles. However, in Brazil, only three species have currently been characterised using molecular methods. This study aimed to bring new insights on the diversity of species of Haemogregarina infecting Brazilian freshwater turtles from Mato Grosso and Goiás states using molecular and morphological tools. (2) Methods: In total, 2 mL of blood was collected, with between two to five blood smears prepared, fixed with absolute methanol, and stained with a 10% Giemsa solution. Blood was stored at −20 °C for molecular analysis targeting the 18S rRNA gene. Fragments of the organs (liver, spleen, heart, and kidney) were separated and stained with hematoxylin-eosin. (3) Results: A total of 40 turtles were screened and hemogregarines were observed in 34 (85%) individuals (3 Podocnemis expansa and 31 Podocnemis unifilis). Haemogregarina embaubali and six new species of Haemogregarina were observed and formally described as follows: H. unifila n. sp., H. rubra n. sp., H. goianensis n. sp., H. araguaiensis n. sp., H. tigrina n. sp., and H. brasiliana n. sp. (4) Conclusions: This study contributes to the diversity and knowledge of Brazilian fresh-water turtle blood parasites, using integrative approaches for diagnosing and characterizing hemoparasites, with the identification of six undescribed species.
Studies on Collaea species (Fabaceae) occurring in Brazil: taxonomic novelties, new interpretations about the leaf of the genus, and new leaf anatomical characters for American Fabaceae
(2022-12-15) da Silva, Marcos José; Fortuna-Perez, Ana Paula [UNESP]; Dos Santos, Igor Soares; Pinto, Rafael Barbosa; Universidade Federal de Goiás (UFG); Universidade Estadual Paulista (UNESP)
Collaea insignis, a new species from Midwestern of Brazil, is described and illustrated from plants recognized as C. speciosa or misinterpreted as C. aschersoniana and C. stenophylla. However, the association of the morphological (e.g., morphology of leaves, flowers and indument types), leaf anatomical (see below), and geographical (allopatric distribution) features led us to recognized such species, and also propose four lectotypes and a synonym within this Collaea group. The leaf anatomy of Collaea species is described here for the first time, and it is noteworthy that all species present stomata in crypts, a rare feature in plants, and not described in the American Fabaceae. Other characters such as indumented leaflets, thick cuticle, palisade parenchyma with 2 or 3 layers of cells, spongy parenchyma with 1–3 layers of loosened cells, cortex in the midrib, petiole, pulvinus and pulvinules with rhombohedral crystals, bicollateral vascular bundles in the midrib and collateral in the petiole and petiole, and common epidermal cells with different shapes and sizes between the abaxial, and adaxial surfaces are also provided for the first time for the genus. In addition to several vegetative and reproductive characters that can be used to separate the new species from morphologically similar congeners. The studied species can be anatomically differentiated by a set of characters such as contour of the midrib, pulvinus, and puvinules, presence or absence of petiole, type of the vascular bundles of the mesophyll, midrib, and pulvinules, type of sheath extension surrounding the vascular bundles in the mesophyll and its direction, as well as the number of parenchyma and collenchyma layers in the cortex of the midrib, pulvinus and pulvinules. In addition to the taxonomic, and macro and micromorphological novelties, new interpretations about the leaf of the genus are proposed.
Clinical outcome of Brazilian patients with non-small cell lung cancer in early stage harboring rare mutations in epidermal growth factor receptor
(2022-01-01) Machado-Rugolo, J. [UNESP]; Baldavira, C. M.; Prieto, T. G.; Olivieri, E. H.R.; Fabro, A. T.; Rainho, C. A. [UNESP]; Castelli, E. C. [UNESP]; Ribolla, P. E.M. [UNESP]; Ab’saber, A. M.; Takagaki, T.; Nagai, M. A.; Capelozzi, V. L.; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); AC Camargo Cancer Center; Instituto do Câncer de São Paulo
The common epidermal growth factor receptor (EGFR) mutations, such as the L858R point mutation in exon 21 and the in-frame deletional mutation in exon 19, have been definitively associated with response to EGFR-tyrosine kinase inhibitors (EGFR-TKI). However, the clinical outcome and response to treatment for many other rarer mutations are still unclear. In this study, we report the results of Brazilian patients in stage IB–IIIA non-small cell lung cancer (NSCLC) following complete resection with minimal residual disease and EGFR mutations treated with adjuvant chemotherapy and/or EGFR-TKIs. The frequency of EGFR mutations was investigated in 70 cases of early stage NSCLC. Mutations in exons 18 and 20, uncommon mutations in exons 19 and 21, as well as in exons 3, 7, 14, 16, 22, 27, and 28, and/or the presence of different mutations in a single tumor (complex mutations) are considered rare. EGFR mutations were detected in 23 tumors (32.9%). Fourteen cases carried rare mutations and were treated with platinum-based chemotherapy and two cases were treated with erlotinib. The clinical outcome is described case by case with references to the literature. Notably, we found two rare EGFR mutations and one of them with an unknown response to chemotherapy and/or EGFR-TKIs. We have provided complementary information concerning the clinical outcome and treatment of patients with early stage NSCLC for several rare EGFR mutations not previously or only rarely reported. Description of cases harboring rare mutations can support the decision-making process in this subset of patients.
Integrative taxonomy in the genus Rhabdias Stiles et Hassall, 1905 from anuran in Brazil, description of two new species and phylogenetic analyses
(2023-04-01) Alcantara, Edna Paulino [UNESP]; Müller, Maria Isabel; Úngari, Leticia Pereira [UNESP]; Ferreira-Silva, Cristiana [UNESP]; Emmerich, Enzo [UNESP]; Giese, Elane Guerreiro; Morais, Drausio Honorio; Santos, André Luiz Quagliatto; O'Dwyer, Lucia Helena [UNESP]; Silva, Reinaldo José [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); Laboratório de Histologia e Embriologia Animal – LHEA; Universidade Federal de Uberlândia (UFU)
About 20 valid species of the genus Rhabdias are known in the Neotropical region. The present study aimed to describe two new species of Rhabdias parasitizing the lungs of Leptodactylus macrosternum and Leptodactylus podicipinus from Brazil. Distinctive characteristics between these species are numerous and based on body size, size of the buccal capsule, shape and size of the oesophagus, and position of the vulva. Molecular data based on ribosomal genes 28S and ITS region and mitochondrial COI of the two species are presented. Molecular analysis and comparison of the partial mitochondrial COI sequence of Rhabdias matogrossensis n. sp. and Rhabdias guaianensis n. sp. revealed a genetic divergence between these new species and the sequences of Rhabdias spp. previously deposited in GenBank. In the phylogenetic analysis, R. matogrossensis n. sp. was grouped with R. breviensis species complex, and R. guaianensis n. sp. was grouped as a sister group of R. cf. stenochepala. This study contributes to improving the diversity of known species of Rhabdias described in Brazilian anurans.
Community structure of polypores (Basidiomycota) in a restored Brazilian Forest
(2022-10-01) Alcantara, Alex A.; Pires, Ricardo M. [UNESP]; Gomes, Eduardo P.C.; Barbosa, Luiz M.; Gugliotta, Adriana M.; Instituto de Pesquisas Ambientais; Universidade Estadual Paulista (UNESP)
Polypores (Basidiomycota) are of great importance in several forest areas since they are fundamental for wood decomposition and nutrient recycling, which is essential for the functioning of ecosystems. This study assessed the polypore community structure in a restoration area and its use as a parameter to monitor restoration. Our study was carried out in Parque Florestal São Marcelo Private Natural Heritage Reserve (RPPN), a protected area of 240 ha, formed in 2002 from reforestation with native species, 13 years after the restoration measures. The polypore community in the area was characterized according to the richness, abundance, and functionality. Results were compared with data from two natural reference areas near the study site and in the same forest type, i.e., riparian forest, but with different land use history. One hundred and eighteen specimens belonging to 31 species were collected; 6.45% were abundant (Funalia rigida and Pycnoporus sanguineus), 12.90% common, 32.26% occasional, and 48.39% rare. Four functional groups based on the species' relative frequency on each substrate were found. Higher frequency of polypores was observed in substrates of smaller diameter (dead branches). The restored area showed a similar richness to the preserved area used as reference. In addition, the polypore community showed a structure similar to well-established areas. This result indicates that the RPPN restoration project was successful and that the polypore community structure can be suitable for evaluating and monitoring regions restored over time.
Intraspecific variation in ultrastructure and secretion of the resin canals in Anacardium humile (Anacardiaceae)
(2023-05-01) de Carvalho, Shelly Favorito [UNESP]; Scudeler, Elton Luiz [UNESP]; dos Santos, Daniela Carvalho [UNESP]; Machado, Silvia Rodrigues [UNESP]; Universidade Estadual Paulista (UNESP)
The study combines a range of light and electron microscopy methods to access variation in secretion and ultrastructure in the secretory canals in the above- and belowground stems of Anacardium humile, which here serves as a model system. The aboveground stem canals show epithelial cells with ultrastructural characteristics typical of cells active in secretion, while in the belowground stems, the subcellular characteristics are typical of cells with low rates of metabolism. The secretory canals of the belowground stems show uniformity in size and shape, a large central vacuole, a cytoplasm reduced to a thin layer at the cell periphery, and a reduced population of organelles. The aboveground stem canals had voluminous nuclei with evident nucleoli, a very dense cytoplasm with free ribosomes, polyribosomes, mitochondria with developed cristae, and ellipsoid plastids with electron-opaque droplets surrounded by a periplastid reticulum. The vacuoles were of different sizes and often had membranous contents and the dictyosomes were very developed with dilated ends to the cisternae, rough endoplasmic reticulum, and numerous vesicles. The results show that particularities in above- and belowground environment have significant implications for ultrastructural morphology and functioning of secretory canals in the stems of A. humile.
Stylosanthes (Leguminosae-Papilionoideae-Dalbergieae) in Northeast Brazil
(2022-01-01) da Silva Ferreira, Jamile Jorge [UNESP]; Perez, Ana Paula Fortuna [UNESP]; Lewis, Gwilym Peter; Silva, Juliana Santos; Universidade Estadual Paulista (UNESP); Accelerated Taxonomy Department; Prog. Pós-graduação em Biodiversidade Vegetal
Stylosanthes, with approximately 50 species, is a pantropical, economically important genus of Leguminosae, with several species having forage potential and soil improvement potential. Thirty-two species have been recorded in Brazil, with the Northeast region being one of the main centers of diversity of the genus in the country. Despite this, no taxonomic work focusing on Stylosanthes has been done for the region. This study presents a taxonomic treatment of Stylosanthes species occurring in the Northeast region of Brazil. Botanical specimens were collected in the field together with visits to the main herbaria in the Northeast (ALCB, HUEFS, HUNEB, HVASF) for specimen analysis. Online consultation of databases in virtual herbaria, analysis of regional flora accounts, assessment of types and discussions with legume experts were also undertaken. Twenty-three taxa were confirmed as occurring in NE Brazil, with six new occurrences recorded for the states of Ceará, Paraíba, Piauí, and Rio Grande do Norte. Species in the studied area are mainly differentiated by their fruit and bract morphology.
Peltate trichomes in the dormant shoot apex of Metrodorea nigra, a Rutaceae species with rhythmic growth
(2023-01-01) Machado, S. R. [UNESP]; de Deus Bento, K. B. [UNESP]; Canaveze, Y.; Rodrigues, T. M. [UNESP]; Universidade Estadual Paulista (UNESP); Federal University of Rio de Janeiro
In Metrodorea nigra, a Rutaceae species with rhythmic growth, the shoot apex in the dormant stage is enclosed by modified stipules. The young organs are fully covered with peltate secretory trichomes, and these structures remain immersed in a hyaline exudate within a hood-shaped structure. Our study focused on the morpho-functional characterization of the peltate trichomes and cytological events associated with secretion. Shoot apices were collected during both dormant and active stages and processed for anatomical, cytochemical and ultrastructural studies. Trichomes initiate secretion early on, remain active throughout leaf development, but collapse as the leaves expand; at which time secretory cavities start differentiation in the mesophyll and secretion increases as the leaf reaches full expansion. The subcellular apparatus of the trichome head cells is consistent with hydrophilic and lipophilic secretion. Secretion involves two vesicle types: the smaller vesicles are PATAg-positive (periodic acid/thiocarbohydrazide/silver proteinate) for carbohydrates and the larger ones are PATAg-negative. In the first phase of secretory activity, the vesicles containing polysaccharides discharge their contents through exocytosis with the secretion accumulating beneath the cuticle, which detaches from the cell wall. Later, a massive discharge of lipophilic substances (lipids and terpenes/phenols) results in their accumulation between the wall and cuticle. Release of the secretions occurs throughout the cuticular microchannels. Continued protection of the leaves throughout shoot development is ensured by replacement of the collapsed secretory trichomes by oil-secreting cavities. Our findings provide new perspectives for understanding secretion regulation in shoot apices of woody species with rhythmic growth.
Near-infrared leaf reflectance modeling of Annona emarginata seedlings for early detection of variations in nitrogen concentration
(2023-02-01) Gomes, Rafaela Lanças [UNESP]; Sousa, Marília Caixeta [UNESP]; Campos, Felipe Girotto [UNESP]; Boaro, Carmen Sílvia Fernandes [UNESP]; de Souza Passos, José Raimundo [UNESP]; Ferreira, Gisela [UNESP]; Universidade Estadual Paulista (UNESP)
Nitrogen (N) monitoring is essential in nurseries to ensure the production of high-quality seedlings. Near-infrared spectroscopy (NIRS) is an instantaneous, nondestructive method to monitor N. Spectral data such as NIRS can also provide the basis for developing a new vegetation spectral index (VSI). Here, we evaluated whether NIRS combined with statistical modeling can accurately detect early variations in N concentration in leaves of young plants of Annona emarginata and developed a new VSI for this task. Plants were grown in a hydroponics system with 0, 2.75, 5.5 or 11 mM N for 45 days. Then we measured gas exchange, chlorophylla fluorescence, and pigments in leaves; analyzed complete leaf nutrients, and recorded spectral data for leaves at 966 to 1685 nm using NIRS. With a statistical learning approach, the dimensionality of the spectral data was reduced, then models were generated using two classes (N deficiency, N) or four classes (0, 2.75, 5.5, 11 mM N). The best combination of techniques for dimensionality reduction and classification, respectively, was stepwise regression (PROC STEPDISC) and linear discriminant function. It was possible to detect N deficiency in seedlings leaves with 100% precision, and the four N concentrations with 93.55% accuracy before photosynthetic damage to the plant occurred. Thereby, NIRS combined with statistical modeling of multidimensional data is effective for detecting N variations in seedlings leaves of A. emarginata.
Roles of Calcium Signaling in Gene Expression and Photosynthetic Acclimatization of Solanum lycopersicum Micro-Tom (MT) after Mechanical Damage
(2022-11-01) Campos, Felipe Girotto [UNESP]; Seixas, Diana Pacheco [UNESP]; Barzotto, Gustavo Ribeiro [UNESP]; Jorge, Letícia Galhardo [UNESP]; Ducatti, Karina Renostro [UNESP]; Ferreira, Gisela [UNESP]; Rodrigues, Tatiane Maria [UNESP]; Silva, Edvaldo Aparecido Amaral da [UNESP]; Boaro, Carmen Sílvia Fernandes [UNESP]; Universidade Estadual Paulista (UNESP)
A momentary increase in cytoplasmic Ca2+ generates an oscillation responsible for the activation of proteins, such as calmodulin and kinases, which interact with reactive oxygen species (ROS) for the transmission of a stress signal. This study investigated the influence of variations in calcium concentrations on plant defense signaling and photosynthetic acclimatization after mechanical damage. Solanum lycopersicum Micro-Tom was grown with 0, 2 and 4 mM Ca2+, with and without mechanical damage. The expression of stress genes was evaluated, along with levels of antioxidant enzymes, hydrogen peroxide, lipid peroxidation, histochemistry, photosynthesis and dry mass of organs. The ROS production generated by mechanical damage was further enhanced by calcium-free conditions due to the inactivation of the oxygen evolution complex, contributing to an increase in reactive species. The results indicated that ROS affected mechanical damage signaling because calcium-free plants exhibited high levels of H2O2 and enhanced expression of kinase and RBOH1 genes, necessary conditions for an efficient response to stress. We conclude that the plants without calcium supply recognized mechanical damage but did not survive. The highest expression of the RBOH1 gene and the accumulation of H2O2 in these plants signaled cell death. Plants grown in the presence of calcium showed higher expression of SlCaM2 and control of H2O2 concentration, thus overcoming the stress caused by mechanical damage, with photosynthetic acclimatization and without damage to dry mass production.
Revisiting the taxonomy of Rhabdias fuelleborni Travassos, 1928 (Nematoda, Rhabdiasidae) with approaches to delimitation of species and notes on molecular phylogeny
(2023-02-01) Müller, Maria Isabel [UNESP]; Morais, Drausio Honorio; da Costa, Lorena Freitas Souza Tavares; de Vasconcelos Melo, Francisco Tiago; Giese, Elane Guerreiro; Ávila, Robson Waldemar; da Silva, Reinaldo José [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); Universidade Federal de Uberlândia (UFU); Universidade Federal do Pará (UFPA); Universidade Federal Rural da Amazônia; Federal University of Ceará (UFC)
Rhabdias fuelleborni was described by Travassos (1926), who identified it in the lungs of Rhinella gr. marina (=Bufo marinus) from São Paulo. Later, Kloss (1971) added new information on the morphology and diagnosis of the species and new data on host and localities. However, its taxonomic status remains uncertain, due in particular to the morphological uniformity of Rhabdias spp., while the original description lacks important morphological data. In the present study, we found and reevaluated the type series of R. fuelleborni, as well as examining fresh material obtained from the type host and type locality, using light and scanning electron microscopy and molecular tools. The type series was deposited by Travassos (1926), and according to museum records the type locality is Paraty, Rio de Janeiro, Brazil. Our morphological and molecular analyses reinforce the taxonomic validity of this species, while molecular phylogeny identified a close relationship between R. fuelleborni and Rhabdias sp.4 and Rhabdias sp. from other studies, and from bufonids. Also R. fuelleborni are sister taxa with R. cf. stenocephala from Leptodactylidae from the northeast of Brazil and R. vencesi from Boophis madagascarariensis, Madagascar, according to mitochondrial COI gene.
Diversity of Haemogregarine Parasites Infecting Brazilian Anurans, with a Description of New Species of Dactylosoma (Apicomplexa: Adeleorina: Dactylosomatidae)
(2022-12-01) Úngari, Letícia Pereira [UNESP]; Netherlands, Edward Charles; Santos, André Luiz Quagliatto; de Alcantara, Edna Paulino [UNESP]; Emmerich, Enzo [UNESP]; da Silva, Reinaldo José [UNESP]; O´Dwyer, Lucia Helena [UNESP]; Universidade Estadual Paulista (UNESP); University of the Free State; Universidade Federal de Uberlândia (UFU)
Purpose: Brazilian anurans are considered the most diverse and species rich around the world. Although in recent years there has been a strong focus on research related to this group of animals, their parasites have not received the same attention. Thus, this study aimed to provide morphological and molecular data on haemogregarines biodiversity infecting Brazilian anurans. Methods: During 2020, 116 anurans were collected from four Brazilian States and their blood and fragment of organs were screened for haemogregarine parasites. Results: From the total, seven (6.03%) animals were found infected with species of Hepatozoon and Dactylosoma. Based on the morphological and molecular analysis, four anurans were found infected with Hepatozoon latrensis. The phylogenetic analysis has shown the isolates from this study grouping with the Brazilian anuran Hepatozoon clade, also with gene similarity ranging from 99.70 to 100% to H. latrensis isolates available on GenBank. Furthermore, three specimens (Trachycephalus typhonius, Leptodactylus latrans, and Rhinella diptycha) were infected with the same species of Dactylosoma (100% genetic similarity), with a genetic similarity of 98.56% to Dactylosoma piperis the only other species described in Brazil. In support of the molecular data, different morphological characters were observed in the blood smears as compared to D. piperis, suggesting that the species of Dactylosoma from the present study infecting three different species of Brazilian anurans is an undescribed species. Conclusion: Thus, this study increases the knowledge of Brazilian anuran blood parasites and demonstrates the importance of using integrative approaches for the diagnosis of haemoparasites.
First characterization of PIWI-interacting RNA clusters in a cichlid fish with a B chromosome
(2022-12-01) Oliveira, Jordana Inácio Nascimento [UNESP]; Cardoso, Adauto Lima [UNESP]; Wolf, Ivan Rodrigo [UNESP]; de Oliveira, Rogério Antônio [UNESP]; Martins, Cesar [UNESP]; Universidade Estadual Paulista (UNESP)
Background: B chromosomes are extra elements found in several eukaryote species. Usually, they do not express a phenotype in the host. However, advances in bioinformatics over the last decades have allowed us to describe several genes and molecular functions related to B chromosomes. These advances enable investigations of the relationship between the B chromosome and the host to understand how this element has been preserved in genomes. However, considering that transposable elements (TEs) are highly abundant in this supernumerary chromosome, there is a lack of knowledge concerning the dynamics of TE control in B-carrying cells. Thus, the present study characterized PIWI-interacting RNA (piRNA) clusters and pathways responsible for silencing the mobilization of TEs in gonads of the cichlid fish Astatotilapia latifasciata carrying the B chromosome. Results: Through small RNA-seq and genome assembly, we predicted and annotated piRNA clusters in the A. latifasciata genome for the first time. We observed that these clusters had biased expression related to sex and the presence of the B chromosome. Furthermore, three piRNA clusters, named curupira, were identified in the B chromosome. Two of them were expressed exclusively in gonads of samples with the B chromosome. The composition of these curupira sequences was derived from LTR, LINE, and DNA elements, representing old and recent transposition events in the A. latifasciata genome and the B chromosome. The presence of the B chromosome also affected the expression of piRNA pathway genes. The mitochondrial cardiolipin hydrolase-like (pld6) gene is present in the B chromosome, as previously reported, and an increase in its expression was detected in gonads with the B chromosome. Conclusions: Due to the high abundance of TEs in the B chromosome, it was possible to investigate the origin of piRNA from these jumping genes. We hypothesize that the B chromosome has evolved its own genomic guardians to prevent uncontrolled TE mobilization. Furthermore, we also detected an expression bias in the presence of the B chromosome over A. latifasciata piRNA clusters and pathway genes.
Ecological Features of Large Neotropical Reservoirs and Its Relation to Health of Cage Reared Fish
(Intech Europe, 2012-01-01) Carvalho, Edmir Daniel [UNESP]; Silva, Reinaldo Jose da [UNESP]; Ramos, Igor Paiva [UNESP]; Kruger Paes, Jaciara Vanessa [UNESP]; Zanatta, Augusto Seawright [UNESP]; Brandao, Heleno [UNESP]; Penha Zica, Erica de Oliveira [UNESP]; Nobile, Andre Batista [UNESP]; Acosta, Aline Angelina [UNESP]; David, Gianmarco Silva [UNESP]; Carvalho, E. D.; David, G. S.; DaSilva, R. J.; Universidade Estadual Paulista (UNESP)

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