Prevalence and molecular characterization of carbapenem-resistant Enterobacterales in patients from a public referral hospital in a non-metropolitan region of Brazil during and post the SARS-CoV-2 pandemic.

Antimicrobial resistance (AMR) poses a global threat, with carbapenem-resistant Enterobacterales (CRE) representing a significant concern due to limited therapeutic options. This study investigated the prevalence of carbapenemase genes in CRE strains isolated from tracheal aspirates of patients at a Brazilian university hospital between January 2020 and August 2023. Bacterial identification was conducted using MALDI-TOF, while carbapenemase genes were detected by qPCR. Demographic and clinical data were collected, and univariate analysis was performed using the chi-square test (p < 0.05). Variables with p ≤ 0.10 were further investigated using the chi-square test for linear trend, along with stratified analysis. Out of 1,133 samples, 111 (9.79%) showed CRE growth, with 46 isolates included in the final sample, predominantly comprising Klebsiella pneumoniae (65.21%) and Serratia marcescens (19.57%). The blaKPC gene was prevalent (78.26%), while blaNDM was detected in 21.74% of cases. The identified population was predominantly male (67.39%), elderly (69.57%), white (56.52%), unmarried (63.04%), and had a low level of education (56.52%). Most patients (69.57%) were in the intensive care unit and remained hospitalized for more than 30 days (76.08%). There was a significant inverse trend between Klebsiella pneumoniae and age (p = 0.045), as well as a direct linear trend between blaNDM and the annual increase in COVID-19 cases in Brazil (p = 0.050). A high probability of finding non-Klebsiella pneumoniae bacteria was observed in patients with prolonged hospital stays, independent of COVID-19 (p = 0.006) and the type of resistance genes (p = 0.020). The persistent prevalence of CRE, especially with blaKPC, underscores the urgency of effective control measures.

Copaiba oil minimizes inflammation and promotes parenchyma re-epithelization in acute allergic asthma model induced by ovalbumin in BALB/c mice.

Introduction: Asthma is a condition of airflow limitation, common throughout the world, with high mortality rates, especially as it still faces some obstacles in its management. As it constitutes a public health challenge, this study aimed to investigate the effect of copaiba oil (e.g., Copaifera langsdorffii), as a treatment resource, at doses of 50 and 100 mg/kg on certain mediators of acute lung inflammation (IL-33, GATA3, FOXP3, STAT3, and TBET) and early mechanisms of lung remodeling (degradation of elastic fiber tissues, collagen deposition, and goblet cell hyperplasia). Methods: Using an ovalbumin-induced acute allergic asthma model in BALB/c mice, we analyzed the inflammatory mediators through immunohistochemistry and the mechanisms of lung remodeling through histopathology, employing orcein, Masson's trichrome, and periodic acid-Schiff staining. Results: Copaiba oil treatment (CO) reduced IL-33 and increased FOXP3 by stimulating the FOXP3/GATA3 and FOXP3/STAT3 pathways. Additionally, it upregulated TBET, suggesting an additional role in controlling GATA3 activity. In the respiratory epithelium, CO decreased the fragmentation of elastic fibers while increasing the deposition of collagen fibers, favoring epithelial restructuring. Simultaneously, CO reduced goblet cell hyperplasia. Discussion: Although additional research is warranted, the demonstrated anti-inflammatory and re-epithelializing action makes CO a viable option in exploring new treatments for acute allergic asthma.

Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR.

Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world's response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to collect and process the samples, and to perform reliable mass diagnosis by RT-qPCR. In particular, time-consuming and high cost of nucleic acid extraction procedures have hampered the diagnosis; moreover, several steps in the routine for the preparation of the material makes the extracted sample susceptible to contamination. Here two rapid nucleic acid extraction reagents were compared as extraction procedures for SARS-CoV-2 detection in clinical samples by singleplex and multiplex RT-qPCR analysis, using different transport media, samples with high and low viral load, and different PCR machines. As observed, rapid nucleic acid extraction procedures can be applied for reliable diagnosis using a TaqMan-based assay, over multiple platforms. Ultimately, prompt RNA extraction may reduce costs with reagents and plastics, the chances of contamination, and the overall time to diagnosis by RT-qPCR.

Adjusting RT-qPCR conditions to avoid unspecific amplification in SARS-CoV-2 diagnosis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and quickly spread around the world, forcing global health authorities to develop protocols for its diagnosis. Here we report dimer formation in the N2 primers-probe set (CDC 2019-nCoV Real-Time RT-PCR) used in the diagnostic routine, and propose alternatives to reduce dimerization events. Late unspecific amplifications were visualized in 56.4% of negative samples and 57.1% of no-template control, but not in positive samples or positive control. In silico analysis and gel electrophoresis confirmed the dimer formation. The RT-qPCR parameters were optimized and the late unspecific amplifications decreased to 11.5% in negative samples and no-template control. The adjustment of PCR parameters was essential to reduce the risk of false-positives results and to avoid inclusive results requiring repeat testing, which increases the costs and generates delays in results or even unnecessary requests for new samples.

Comparative analysis for strength serum sodium and potassium in three different methods: Flame photometry, ion-selective electrode (ISE) and colorimetric enzymatic.

BACKGROUND: Hydroelectrolytic disorders are common in clinical situations and may be harmful to the patient, especially those involving plasma sodium and potassium dosages. Among the possible methods for the dosages are flame photometry, ion-selective electrode (ISE) and colorimetric enzymatic method. METHODS: We analyzed 175 samples in the three different methods cited from patients attending the laboratory of the University Hospital of the Federal University of Juiz de Fora. The values obtained were statistically treated using SPSS 19.0 software. The present study aims to evaluate the impact of the use of these different methods in the determination of plasma sodium and potassium. RESULTS: The averages obtained for sodium and potassium measurements by flame photometry were similar (P > .05) to the means obtained for the two electrolytes by ISE. The averages obtained by the colorimetric enzymatic method presented statistical difference in relation to ISE, both for sodium and potassium. In the correlation analysis, both flame photometry and colorimetric enzymatic showed a strong correlation with the ISE method for both dosages. CONCLUSION: At the first time in the same work sodium and potassium were analyzed by three different methods and the results allowed us to conclude that the methods showed a positive and strong correlation, and can be applied in the clinical routine.

PCR Amplification of DNA Sequence of Bovine Papillomavirus Type 2 in Urinary Bladder of Cattle with Enzootic Hematuria in Espirito Santo, Brazil

The goal of this study was to investigate the presence of bovine papillomavirus type 2 (BPV-2) genomic DNA using polymerase chain reaction (PCR) assays, in the urinary bladder of cattle with bovine enzootic hematuria (BEH), in the neoplastic and non-neoplastic lesions in Espírito Santo, Brazil. Thus, 46 bladders with gross BEH lesions were collected from adult cattle at slaughterhouses. The samples were divided into two portions: one for use in histopathological analysis and the other one for PCR. Microscopic analysis showed that 56.52% of the bladders presented neoplastic lesions, of which 50% were benign, 23.08% were malignant and 26.92% presented both types of neoplasms. The remaining 43.48% of the bladders did not show any neoplasms. Presence of BPV-2 was confirmed in 43.48% of the bladders: 65% of these were found in neoplastic lesions and 35% in non-neoplastic lesions. Among the neoplastic lesions, detection of BPV-2 was more frequent in benign lesions (53.84%), followed by malignant lesions (23.08%) and by those presenting both benign and malignant lesions (23.08%). These results confirm the presence of BPV-2 in both neoplastic and non-neoplastic conditions in urinary bladders of cattle with enzootic hematuria from the studied geographic area.(AU)

PCR Amplification of DNA Sequence of Bovine Papillomavirus Type 2 in Urinary Bladder of Cattle with Enzootic Hematuria in Espirito Santo, Brazil

The goal of this study was to investigate the presence of bovine papillomavirus type 2 (BPV-2) genomic DNA using polymerase chain reaction (PCR) assays, in the urinary bladder of cattle with bovine enzootic hematuria (BEH), in the neoplastic and non-neoplastic lesions in Espírito Santo, Brazil. Thus, 46 bladders with gross BEH lesions were collected from adult cattle at slaughterhouses. The samples were divided into two portions: one for use in histopathological analysis and the other one for PCR. Microscopic analysis showed that 56.52% of the bladders presented neoplastic lesions, of which 50% were benign, 23.08% were malignant and 26.92% presented both types of neoplasms. The remaining 43.48% of the bladders did not show any neoplasms. Presence of BPV-2 was confirmed in 43.48% of the bladders: 65% of these were found in neoplastic lesions and 35% in non-neoplastic lesions. Among the neoplastic lesions, detection of BPV-2 was more frequent in benign lesions (53.84%), followed by malignant lesions (23.08%) and by those presenting both benign and malignant lesions (23.08%). These results confirm the presence of BPV-2 in both neoplastic and non-neoplastic conditions in urinary bladders of cattle with enzootic hematuria from the studied geographic area.

Fasciola hepatica em ovinos, caprinos e bubalinos em municípios do sul do Espírito Santo / Fasciola hepatica in sheep, goats and buffaloes in southern municipalities of Espírito Santo state

O objetivo deste trabalho foi registrar a presença de Fasciola hepatica em ovinos, caprinos e bubalinos em dez municípios do sul do Espírito Santo e relacionar fatores determinantes da doença na região. Um total de 34 propriedades de ovinos, caprinos e bubalinos foi visitado coletando-se amostras fecais dessas espécies para procedimento da técnica de sedimentação. Um total de 58 (13,68%) amostras de ovinos, 76 (21,78%) amostras de caprinos e 10 (23,81%) amostras de bubalinos foram positivas para ovos de F. hepatica. Todos os moluscos coletados foram identificados como pertencentes à espécie Lymnaea columella. Notou-se que os municípios da região sul do Espírito Santo apresentam altas taxas de F. hepatica em ovinos, caprinos e bubalinos, tornando essa região um grande foco de dispersão da parasitose, e que a ocorrência de casos anteriores de fasciolose nas propriedades ficou caracterizada como fator de risco epidemiológico para a doença.

The aim of this study was to record the presence of Fasciola hepatica in sheep, goats and buffaloes in ten southern municipalities of the Espírito Santo state and to relate determinant factors of the disease in this region. A total of 34 properties in sheep, goats and buffaloes was visited by collecting fecal samples of these species to the sedimentation procedure. A total of 58 (13.68%) samples of sheep, 76 (21.78%) samples of goats and 10 (23.81%) samples of buffaloes was positive for F. hepatica eggs. Molluscs collected were all identified as Lymnaea columella. We noted that the municipalities in the south of Espírito Santo have high rates of F. hepatica in sheep, goats and buffaloes, which makes this region a major focus of dispersal of the parasite, and that the occurrence of previous cases of fascioliasis in the properties was characterized as an epidemiological risk factor for the disease.