SARS-CoV-2 intralineage variation and temporal patterns of COVID-19 risk factors in three cities of southeastern Brazil: Age, sex, and race.

The Santo André Regional Center from Adolfo Lutz Institute evaluated 91 537 samples by reverse transcription-polymerase chain reaction (RT-PCR) to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from March 2020 to April 2021. The age, sex, and race of patients from three cities in southeastern Brazil, namely São Bernardo do Campo (SBC), Diadema, and Mauá were assessed in association to the rate of positive results using generalized linear models. Circulating lineages were obtained from GISAID and intralineage genetic variation was investigated employing Lasergene software. A declining number of reported cases around October to November 2020 separate two epidemic waves in the three cities. Mauá differed by the highest positive RT-PCR scores in January and February. GISAID classification of 38 SARS-CoV-2 complete genomic sequences showed the circulation of lineages P.1, B.1.1.28, P.2, B.1.1.332; P.1, P.2, B.1.1.28, B.1.1.33; and P.1, P.2 in SBC, Diadema and Mauá, respectively. Intralineage variation revealed a significant amino-acid substitution in the ORF3a encoding protein (A33S) present in four out of six (67%) P.1 Mauá isolates. As ORF3a encodes a nonselective Ca2+ permeable cation channel, supposed to interfere in airway homeostasis, specific mutations could increase its pathogenic effect resulting in a higher number of symptomatic individuals explaining why the second wave was more intense in Mauá city.

Viability assessment of Mycobacterium tuberculosis complex in OMNIgene • SPUTUM reagent using the BACTEC MGIT 960 System and Xpert MTB/RIF assay.

The World Health Organization advocates that sputum specimens submitted to tuberculosis (TB) diagnostic should be processed within 48 h after collection and be stored under cooling. We aimed to assess the performance of OMNIgene • SPUTUM reagent in maintaining viable specimens of Mycobacterium tuberculosis complex (MTBC) during transportation of sputum samples without refrigeration, in comparison to the standard protocol of the National TB Control Program. Sputum samples obtained in southeastern Brazil (June 2017 to July 2018) from 100 sequential patients with positive acid-fast bacillus smear microscopy were divided into two portions. Portion 1 continued to be cooled (standard protocol, STA), but portion 2 was added to OMNIgene • SPUTUM reagent (alternative protocol, OMS) until concomitant further processing. Both portions of all samples were cultured using MGIT and tested by Xpert MTB/RIF assay. Growth of MTBC in the first 42 days was detected in 96% of the cultures under the STA and 88% under the OMS. Intervals between processing and detecting MTBC growth in the two portions significantly differed (p = 0.0001). Portions under the two protocols showed similar results in the MTBC detection by Xpert assay and culture contamination by non-MTBC. The OMNIgene reagent liquefies and decontaminates sputum leading to a decrease in processing time. Although there was a small delay in mycobacterial growth, the OMNIgene reagent can be useful in specimens transported from collection sites over a long distance to centralized testing centers, maintaining viable MTBC for at least 8 days at room temperature.

Epidemiological and genetic aspects of the largest dengue outbreak recorded in 2015 in Southeastern Brazil.

Dengue virus, the etiological agent of dengue fever (DF) occurs in four genetically distinct serotypes (DENV1-4), being transmitted by female Aedes mosquitoes. DF incidence is increasing in Brazil, following vector dispersal, proliferation and DENV serotypes introduction, co-circulation and substitution. Medium- and small-sized cities in Sao Paulo State, such as Marilia (Midwest region), have been affected by huge epidemics. To understand the evolution of DENV epidemics in medium-sized cities, in this study a historical data on DENV incidence (2000-2015) in Marilia, was evaluated. Previous studies disclosed regional and specific DF outcomes associated with 2007 outbreak in that city, when co-circulating DENV1 and DENV3 presented different hematological profiles. In this study, characteristics of 2007 DENV epidemics were compared to the epidemiological, hematological and demographic outlines of the major outbreak of DENV1 in Marilia in 2015. DENV1 genetic diversity was assessed through capsid and pre-membrane junction encoding gene (CprM) sequencing. The results revealed circulation of DENV1 serotype from 2007 to 2015, with epidemics occurring every three-years until 2013 and then, increasing yearly. There were significant differences in hematological profiles of DENV1 patients between 2015 and 2007. CprM showed DENV1 genetic variability in 2015, contrasting with the unique sequence pattern in 2007. These results reinforce the regional and temporal characteristics of DENV epidemics that need local public health research to improve care for people and to limit the spread of new serotypes/genotypes to uninfected areas.

Being a bright snake: testing aposematism and mimicry in a neotropical forest

Based on color patterns and behavioral similarities, venomous coral snake Micrurus corallinus (Elapidae) may act as a model for two polymorphic species, Erythrolamprus aesculapii (Dipsadidae) and Micrurus decoratus (Elapidae). Plasticine replicas were used to investigate the aposematism of these coloration patterns and whether these species may be part of mimetic complexes in two Atlantic Forest localities in Southeast Brazil. Coral replicas were more avoided when set upon a white background, evincing that the pattern may act aposematically in contrast with light substrates. Birds attacked all four patterns equally during the mimicry experiments. Birds of prey, known to be effective in predating snakes, are quite abundant in the study areas, which may have led to this lack of avoidance. Accordingly, they predated more adult‐sized replicas, which could be more dangerous. Interestingly, opossum avoided the Micrurus corallinus and Erythrolamprus aesculapii replicas that resembled the model. This suggests that opportunistic predators, as the opossum may be important selective agents in mimicry complexes.

Epidemiology and genetic screening of diarrheagenic Escherichia coli among symptomatic and asymptomatic children.

PURPOSE: Enteropathogens are frequently associated with diarrheal disease. Knowledge of their etiology and epidemiology is essential for the prevention and control of the sickness. This study describes the microbiological and epidemiological features of diarrheal disease in 197 symptomatic and 223 asymptomatic under-five-year-old children from southeastern Brazil, between January 2015 and September 2016. METHODS: Isolation of Escherichia coli, Salmonella, Shigella and Campylobacter was realized by culture. E. coli strains were screened by multiplex PCR, PFGE and O:H serotyping. Antimicrobial susceptibility testing was also performed. RESULTS: Most of the 127 enteropathogens isolated were diarrheagenic E. coli (96.1 %), with predominance of several serotypes of enteropathogenic E. coli (EPEC) and enteroaggregative E. coli (EAEC). Age, sex, rotavirus vaccination, recent use of antibiotics and previous contact with pets, were factors that revealed no significant effects on the probability of infection by the predominant pathogens. Even so, higher incomes could be related to a lesser chance of testing positive for EPEC. Evidence of possible EAEC clonal spread was detected, as well as genetic similarity among strains from both symptomatic and asymptomatic children. Resistance to antimicrobial agents was more pronounced among EAEC than EPEC. CONCLUSION: The occurrence of genetically similar diarrheagenic E. coli in both groups of children, likewise resistant to these agents, underscores the importance of establishing strategies for the prevention of outbreaks, especially among low-income households.

Co-circulating serotypes in a dengue fever outbreak: Differential hematological profiles and phylogenetic relationships among viruses.

BACKGROUND: Dengue virus, represented by four distinct, genetically diverse serotypes, is the etiologic agent of asymptomatic to severe hemorrhagic diseases. The spatiotemporal dynamics of dengue serotypes and its association to specific diseases vary among the different regions worldwide. By 2007, and in São Paulo State, Brazil, dengue-case concentration in urban centers had changed to increased incidence in small- and medium-sized towns, the case of Marília. OBJECTIVES: The aim of this article was to distinguish dengue serotypes circulating during the 2007 Marília outbreak and define their association to demographic and hematological patient profiles, as well as the phylogenetic relationships among the different viruses. STUDY DESIGN: PCR amplicons corresponding to the junction of capsid and dengue pre-membrane encoding genes, obtained from dengue serologically positive patients, were sequenced. Hematological and demographic data of patients with different Dengue serotypes were evaluated by univariate and bivariate statistics. Dengue PCR sequences were used in phylogenetic relationships analyzed for maximum parsimony. RESULTS: Molecular typing confirmed co-circulation of the dengue serotypes 1 (DENV1) and 3 (DENV3), which presented divergent correlation patterns with regard to hematological descriptors. The increase in atypical lymphocytes, a likely indication of virus load, could be significantly associated to a decrease in leukocyte counts in the DENV3 group and platelet in the DENV1. Phylogenetic reconstitution revealed the introduction of DENV1 from northern Brazil and local divergence of DENV3 by either microevolution or viral introduction from other geographical regions or both. CONCLUSIONS: Dengue dynamics showed regional molecular-epidemiologic specificity, which has important implications for introduction of vaccines, disease management, and transmission control.

Maintenance of demographic and hematological profiles in a long-lasting dengue fever outbreak: implications for management.

BACKGROUND: Dengue fever (DF) outbreaks present regionally specific epidemiological and clinical characteristics. In certain medium-sized cities (100 000-250 000 inhabitants) of São Paulo State, Brazil, and after reaching an incidence of 150 cases/100 000 inhabitants ("epidemiological threshold"), clinical diagnosis indicated dengue virus (DENV) infection. During this period, other seasonally infectious diseases with symptoms and physical signs mimicking DF can simultaneously occur, with the consequential overcrowding of health care facilities as the principal drawbacks. Confirmation of clinical diagnosis of DF with serological tests may help in avoiding faulty diagnosis in patients, who might later undergo dengue hemorrhagic fever (DHF) and the dengue-shock syndrome (DSS). Furthermore, demographic and hematological profiles of patients are useful in detecting specific early characteristics associated to DF, DHF and DSS. METHODS: From March to June, 2007, 456 patients from Marilia in northwest São Paulo State who had only been diagnosed for DF by clinical criteria, underwent serologic testing for non-structural 1 (NS1) DENV antigens. Individual results were used in comparative analysis according to demographic (gender, age) and hematological (leukocyte and platelet counts, percentage of atypical lymphocytes) profiles. Temporal patterns were evaluated by subdividing data according to time of initial attendance, using recorded variables as predictors of DENV infection in logistic regression models and ROC curves. RESULTS: Serologic DENV detection was positive in 70.6 % of the patients. Lower leukocyte and platelet counts were the most important factors in predicting DENV infection (respective medians DENV + = 3 715 cells/ml and DENV- = 6 760 cells/ml, and DENV + = 134 896 cells/ml and DENV- = 223 872 cells/ml). Furthermore, all demographic and hematological profiles presented a conservative temporal pattern throughout this long-lasting outbreak. CONCLUSIONS: As consistency throughout the epidemic facilitated defining the conservation pattern throughout the early stages, this was useful for improving management during the remaining period.

The world's biogeographical regions revisited: global patterns of endemism in Tipulidae (Diptera).

This paper explores the distributional data of 4,224 Tipulidae (Insecta: Diptera) species to search for endemism patterns in a worldwide scale and to test the extent to which the global patterns of endemism of the group fit into previously proposed regionalization schemes, particularly Wallace's system and recent revisions of it. Large scale areas of endemism are assessed using the grid-based method implemented in VNDM. VNDM depends on the prior definition of the grid size for analysis, but a criterion for choosing beforehand a particular grid size is not clear. The same holds for the choice of the level of similarity in species composition selected for the calculation of consensus areas. In our study, we developed a methodological approach that helped defining objective criteria for choosing suitable values for these critical variables. Large-scale areas of endemism around the globe are identified and ranked according to endemicity levels: 1--West Palaearctic, 2--Nearctic, 3--East Palaearctic-Oriental, 4--West North America, 5--Australia, 6--Neotropical, 7--Sub-Saharan Africa, 8--Palaearctic, and 9--Middle East. Our main conclusion is that there are still some limitations in applying biogeographical classifications proposed mostly on the basis of vertebrate distribution to other taxonomic groups, such as the Tipulidae. While there is a general congruence of the broad-scale areas of endemism of tipulids with previously proposed regionalization schemes, for some areas, the sharpness of boundaries between traditional regions is not so acute, due to a great level of overlap of part of its biotic elements.

Different risk factors influence peptic ulcer disease development in a Brazilian population.

AIM: To investigate age, sex, histopathology and Helicobacter pylori (H. pylori) status, as risk factors for gastroduodenal disease outcome in Brazilian dyspeptic patients. METHODS: From all 1466 consecutive dyspeptic patients submitted to upper gastroscopy at Hospital das Clinicas of Marilia, antral biopsy specimens were obtained and subjected to histopathology and H. pylori diagnosis. All patients presenting chronic gastritis (CG) and peptic ulcer (PU) disease localized in the stomach, gastric ulcer (GU) and/or duodenal ulcer (DU) were included in the study. Gastric biopsies (n = 668) positive for H. pylori by rapid urease test were investigated for vacuolating cytotoxin A (vacA) medium (m) region mosaicism by polymerase chain reaction. Logistic regression analysis was performed to verify the association of age, sex, histopathologic alterations, H. pylori diagnosis and vacA m region mosaicism with the incidence of DU, GU and CG in patients. RESULTS: Of 1466 patients submitted to endoscopy, 1060 (72.3%) presented CG [male/female = 506/554; mean age (year) ± SD = 51.2 ± 17.81], 88 (6.0%) presented DU [male/female = 54/34; mean age (year) ± SD = 51.4 ± 17.14], and 75 (5.1%) presented GU [male/female = 54/21; mean age (year) ± SD = 51.3 ± 17.12] and were included in the comparative analysis. Sex and age showed no detectable effect on CG incidence (overall χ² = 2.1, P = 0.3423). Sex [Odds ratios (OR) = 1.8631, P = 0.0058] but not age (OR = 0.9929, P = 0.2699) was associated with DU and both parameters had a highly significant effect on GU (overall χ² = 30.5, P < 0.0001). The histopathological results showed a significant contribution of ageing for both atrophy (OR = 1.0297, P < 0.0001) and intestinal metaplasia (OR = 1.0520, P < 0.0001). Presence of H. pylori was significantly associated with decreasing age (OR = 0.9827, P < 0.0001) and with the incidence of DU (OR = 3.6077, P < 0.0001). The prevalence of m1 in DU was statistically significant (OR = 2.3563, P = 0.0018) but not in CG (OR = 2.678, P = 0.0863) and GU (OR = 1.520, P= 0.2863). CONCLUSION: In our population, male gender was a risk factor for PU; ageing for GU, atrophy and metaplasia; and H. pylori of vacA m1 genotype for DU.

Acidentes provocados por Millipede com manifestações dermatológicas: relatos de dois casos / Accidents provoked by Millipede with dermatological manifestations: report of two cases

Mediante a descrição de dois acidentes provocados por artrópodas (Millipede), os autores comentam essa forma de acidente, descrevem o agente envolvido, seu mecanismo de agressão e apresentam referências bibliográficas sobre o tema, pouco estudado no Brasil