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This methodological study outlines effective and standardized procedures for obtaining high-quality chromosomal preparations in birds, which display complex karyotypes with a large number of microchromosomes. We describe every stage from sample obtaining and culture setup to the successful acquisition of metaphase chromosomes appropriate for in-depth analysis by combining three main methodologies (fibroblast, lymphocyte, and bone marrow cultures) into a coherent and methodical workflow. Furthermore, we offer thorough troubleshooting instructions to tackle typical problems, such as low mitotic indices, poor chromosome spreading, contamination, and suboptimal chromosomal morphology. We also focused on repeatability and adaptation across several bird taxa, taking into consideration species-specific differences in their body sizes, cell development, and chromosomal behavior, extending beyond standard procedure improvement. The incorporation of diverse methodologies into a unified framework offers avian cytogenetics researchers a readily available and practical workflow. Ultimately, our approach enhances the integration of cytogenetic data with novel genomic resources, facilitates the re-evaluation of chromosomal counts, and enables more precise karyotype characterization. This integration is crucial for advancing our understanding of chromosomal evolution, avian genome architecture, and the broader mechanisms underlying avian speciation and diversification.
The sand fly genus Trichophoromyia Barretto, 1962 is one of the most diverse inthe subfamily Phlebotominae. The taxonomy and systematics of this group is complex due toboth a high similarity among species and unclear relationships among other sand fly groupswithin the subtribe Psychodopygina Galati, 1995. Despite their great relevance as vectors of Leishmania spp., few studies have explored the usefulness of molecular markers in studyingthe diversity of this group. Here, we evaluated the use of barcode sequences of the cytochrome coxidasesubunit I gene (COI) for identifying several Trichophoromyia spp., by inferring intra- andinterspecific genetic distances, in addition to performing a set of several single-locus speciesdelimitation approaches using discovery methods. Moreover, we employed a multilocusdataset of four independent molecular markers (COI , ITS2 , 28S and PARA) to infer thephylogenetic species tree, estimate divergence times and delimit species under a validationmodel. The phylogenetic inferences confirmed the paraphyly of Trichophoromyia and Nyssomy ia Barretto, 1962. Thus, two new genera, named Reburrus gen. nov. and Shawmyia gen. nov., were proposed to accommodate sand flyspecies that did not fit in the aforementioned groups. Additionally, a new subgenus wasproposed: Trichophoromyia (Dilermandomyia) subg. nov., containing most speciesof Trichophoromyia . A recent speciation history was also estimated, with most of the speciesstudied diversifying during the Pleistocene. However, our dataset was insufficient to fullyresolve relationships within Trichophoromyia (Dilermandomyia) subg. nov. Many speciesshowed paraphyletic patterns in the gene trees, and some could not be reliably identified anddelimited using both COI barcodes and multilocus tools. The sand fly genus Trichophoromyia exhibits a complex diversification history.Our phylogenetic inference and morphological observations of Nyssomyia and Trichophoromy ia, allowed us to propose new groups for the Psychodopygina subtribe. However, theprevalence of species-level paraphyletic patterns for Trichophoromyia (Dilermandomyia) subg.nov., showed that further assessment of this group requires a broader locus sampling combined with detailed morphological analysis.
The epidemiology of human T-lymphotropic virus (HTLV) among sexual minority cisgender women (SMCW) remains poorly understood globally. We conducted a cross-sectional study of 251 SMCW in Belém, Brazilian Amazon, between March 2023 and April 2025. Participants underwent serological screening (ELISA) with molecular confirmation (qPCR). The prevalence of HTLV infection was 0.4% (1/251), with HTLV-2 identified in a 46-year-old asymptomatic bisexual woman with previous male partners and a current female partner. Serological testing showed strong ELISA reactivity (OD ratio 3.8), and qPCR confirmed HTLV-2 with a Ct value of 28.4 for the pol gene, while HTLV-1 remained negative. This represents the first documented HTLV-2 case in SMCW in the Amazon region, demonstrating viral circulation in an understudied population. Despite the low prevalence, comparable to the general population, these findings highlight the need for inclusive epidemiological surveillance and culturally sensitive prevention strategies for LGBTQIA+ communities in endemic regions.
Purpose: This systematic review aimed to compile studies discussing the influence of spiritual interventions on the recovery and quality of life of patients with CKD, compared to those receiving only conventional treatment. Methods: The review was conducted through searches in the SciELO, PubMed, Cochrane, LILACS and BVS databases, covering articles published between 2015 and 2025. Risk of bias was assessed using Rob 2.0 for randomized clinical trials, ROBINS-I for non-randomized trials, and JBI for observational studies. The quality of evidence was rated according to GRADE system criteria, based on the classification of the outcomes. Results: The analysis of the included studies revealed a positive relationship between spiritual practices and quality of life in CKD patients. However, there were variations in the instruments used and the population studied. Most studies presented a high risk of bias and low methodological precision. Conclusion: There is a growing need for more rigorous and standardized research on the subject in order to support the therapeutic use of spiritual practices that positively impact the quality of life of patients with CKD.
This study analyzed temporal trends and sociodemographic and territorial inequalities in police records of rape and rape of vulnerable persons in Brazil from 2021 to 2025. An ecological, descriptive, longitudinal study was conducted using data from the Brazilian Public Security Yearbooks published between 2022 and 2026. Absolute numbers, rates per 100,000 inhabitants, annual changes, sex, macroregion, federative unit, race/skin color, age group, relationship to the perpetrator, and place of occurrence were examined. Data from 2020 were used exclusively as the baseline for the 2021 variation. Records increased from 68,885 in 2021 to 84,388 in 2025, a 22.5% rise, after peaking at 88,843 in 2024. Female victims predominated, and more than three quarters of the records involved people aged 0–17 years. Black victims were the majority among cases with valid race/skin color information. The home remained the main setting, and family members or other acquaintances accounted for most reported perpetrators. The North, South, and Central-West showed higher rates, although with different trajectories. The findings demonstrate persistence, concentration among children and adolescents, and sex, racial, and territorial inequalities, requiring intersectoral prevention, improved records, protection within close relationships, and stronger care and accountability networks.
