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The reservoirs for NDM-producing Enterobacterales are increasing, not only in hospitals, but also in the environment and in the community, challenging the therapeutic efficacy of carbapenems. We aimed to characterize an isolate of Escherichia coli harboring the bla gene recovered from the bloodstream of a penguin (Spheniscus magellanicus) in Southern Brazil. A total of 74 bacterial isolates recovered from arterial blood samples from dead birds were submitted to species identification and antibiotic susceptibility evaluation. One isolate presented resistance to carbapenems (E. coli 89PenNDM) and proved to harbor the bla gene by multiplex high-resolution melting real-time PCR (PCR-HRM). Conjugation experiments indicated that the bla was transmissible to E. coli J53. Whole genome sequencing (WGS) confirmed the presence of the bla gene in a conjugative plasmid (IncA/C plasmid) in both the E. coli 89PenNDM and its transconjugants. The isolate was classified as ST 156 and many other resistance genes (e.g., sul1, sul,2, strA, floR, tet(A)) were identified, all carried in the same IncA/C2 plasmid. This is the first report of bla -producing E. coli isolated from a penguin in the Brazilian seacoast. The presence of a carbapenemase gene in wildlife animals is of concern as they may become reservoirs of multidrug-resistant bacteria and disseminate them to the environment.

The reservoirs for NDM-producing Enterobacterales are increasing, not only in hospitals, but also in the environment and in the community, challenging the therapeutic efficacy of carbapenems. We aimed to characterize an isolate of Escherichia coli harboring the bla NDM-1 gene recovered from the bloodstream of a penguin ( Spheniscus magellanicus ) in Southern Brazil. A total of 74 bacterial isolates recovered from arterial blood samples from dead birds were submitted to species identification and antibiotic susceptibility evaluation. One isolate presented resistance to carbapenems ( E. coli 89PenNDM) and proved to harbor the bla NDM-1 gene by multiplex high-resolution melting real-time PCR (PCR-HRM). Conjugation experiments indicated that the bla NDM-1 was transmissible to E. coli J53. Whole genome sequencing (WGS) confirmed the presence of the bla NDM-1 gene in a conjugative plasmid (IncA/C 2 plasmid) in both the E. coli 89PenNDM and its transconjugants. The isolate was classified as ST 156 and many other resistance genes (e.g., sul1, sul,2, strA, floR, tet(A)) were identified, all carried in the same IncA/C2 plasmid. This is the first report of bla NDM-1 -producing E. coli isolated from a penguin in the Brazilian seacoast. The presence of a carbapenemase gene in wildlife animals is of concern as they may become reservoirs of multidrug-resistant bacteria and disseminate them to the environment.

The red-tailed Amazon parrot (Amazona brasiliensis) is a threatened species of psittacine bird that inhabit coastal regions of Brazil. In view of the threat of this species, the aim of this study was to perform a health evaluation in wild nestlings in Rasa Island, determining the prevalence of enterobacteria and infectious agents according to type of nest. Blood samples were collected from 64 birds and evaluated for antibodies of Chlamydia psittaci by commercial dot-blot ELISA. Cloacal and oropharyngeal swabs samples were collected from 23 birds from artificial wooden nests, 15 birds from PVC nests and 2 birds from natural nests for microbiological analysis. Swab samples were collected from 58 parrots for C. psittaci detection by PCR and from 50 nestlings for Avian Influenza, Newcastle Disease and West Nile viruses’ detection analysis by real-time RT-PCR. Ten bacterial genera and 17 species were identified, and the most prevalent were Escherichia coli and Klebsiella oxytoca. There was no influence of the type of nest in the nestlings’ microbiota. All samples tested by ELISA and PCR were negative. There is currently insufficient information available about the health of A. brasiliensis and data of this study provide a reference point for future evaluations and aid in conservation plans.

Research background. Extracts from grape pomace, including the wine, show many biological effects such as antioxidant and anti-inflammatory activities. Unfortunately, winemakers discard the bagasse, so the waste is not exploited, although it contains bioactive compounds with antioxidant and anti-inflammatory properties. The work aims to analyze the hydroethanolic extract of peels from Vitis labrusca agro-industrial waste and to evaluate its antinociceptive and anti-inflammatory properties. This study is relevant for reusing a residue and adding value to the grape economic chain. Experimental approach. A representative sample of pomace was obtained and the peels were used to produce the extract. The phenolic compounds were determined by mass spectrometry in multiple reaction monitoring mode and Folin-Ciocalteu colorimetric method, using gallic acid as standard. The biological analyses were carried out using mice orally treated with crude extract at doses of 30, 100 and 300 mg/kg. We evaluated mechanical hyperalgesia by the von Frey method, thermal heat hyperalgesia using a hot plate at 55 °C, paw edema using a Vernier caliper, and neutrophil recruitment by measurement of myeloperoxidase activity. The nephrotoxicity and hepatotoxicity were evaluated by biochemical analyses using blood samples that were collected after the Vitis labrusca administration. Results and conclusions. In all wet winemaking residues peel mass fraction was 75 %, and in dry residues 59 %. We identified nine anthocyanins (3-O-glucosides: peonidin, delphinidin, petunidin and malvidin; 3-p-coumaroyl-glucosides: cyanidin, peonidin, petunidin and malvidin, and malvidin-3,5-diglucoside), five flavonoids (apigenin-7-glucoside, luteolin-7-glucoside, quercetin-3-galactoside, isorhamnetin-3-glucoside and myricetin-3-rutinoside), and mass fraction of phenolic compounds, expressed as gallic acid equivalents, was 26.62 mg/g. In vivo assays showed that Vitis labrusca extract at mass fractions 100 and 300 mg/kg reduced carrageenan-induced mechanical and thermal hyperalgesia, 50 % of the paw edema, and neutrophil recruitment. In addition, there were no indications of nephrotoxicity and hepatotoxicity. Our extract obtained from winemaking residue has analgesic and anti-inflammatory properties, related at least in part to the presence of phenolic compounds, and it is not toxic to renal and hepatic tissues. Novelty and scientific contribution. This bio-product can be used as an alternative to synthetic anti-inflammatory agents with the same pharmacological potential and fewer side effects. We demonstrated that Vitis labrusca winemaking waste can be used for the production of antinociceptive and anti-inflammatory products (nutraceutical, pharmaceutical and cosmetics) without toxicity, contributing to the environmental economy.

This study optimized the extraction process of yellow passion fruit seed waste, using dynamic maceration (DME) and ultrasound-assisted extraction (UAE) methods. Hydroethanolic solvent, time and temperature were studied. The extracts were evaluated for antioxidant activity by DPPH method and Fe (II) ion chelating ability. In terms of solvent optimization, experiments performed by DME, and ethanol: water with the ratio of 80:20 (v/v) indicated the best antioxidant responses. A central composite rotatable design was used to optimize the DME parameters (time and temperature). The experimental condition that provided the extract with the highest antioxidant potential was 41 min and /45 °C. The best condition for the UAE was 15 min and 45 °C. The results of antioxidant activity and yield obtained by UAE were superior to those of DME. The methods used for antioxidant activity estimation were ABTS + radical scavenging, Fe (II) Ion Chelating Ability, DPPH free radical scavenging, phenolic and flavonoid compounds. Mass spectrometry was used to characterize the extracts. Fatty acids, were identified by the full scan method and phenolic acids, flavonoids and stilbenes by the multiple reaction monitoring method. The photoprotective activity of the optimized extracts was analyzed by determining the sun protection factor (SPF). The extracts obtained by DME and UAE had the SPF of 18.22 and 11.68, respectively, suggesting their incorporation in dermo-cosmetic formulations. The extract obtained by UAE demonstrated an antiproliferative effect on the SIHA cell line and cell division stimulating effect on the HuH7.5 cell line. The results showed that the extract from passion fruit seeds which are considered waste has high antioxidant potential and compounds of biological interest, which are necessary for further studies aiming at its pharmacological application. Graphical Abstract

The aim of this study was to determine the apparent digestibility coefficients and digestible values of crude protein, ethereal extract, gross energy and dry matter of mulberry leaf meal (MLM) (Morus alba L.) as Nile tilapia (Oreochromis niloticus) feed. A total of 135 Nile tilapia juveniles were used, and the indirect methodology (Cr2O3) was applied for digestibility determinations. Mulberry leave meal presented good apparent digestible coefficients of protein, ethereal extract and energy with respective values of 0.94, 0.58 and 0.39. The mulberry leave meal thus comprises adequate digestible protein and digestible energy values, similar or better than other leafy foods, presenting potential for inclusion in Nile tilapia diets.

Background Spirometry and conventional chest x-ray have limitations in investigating early emphysema, while computed tomography, the reference imaging method in this context, is not part of routine patient care due to its higher radiation dose. In this work, we investigated a novel low-dose imaging modality, dark-field chest x-ray, for the evaluation of emphysema in patients with alpha1-antitrypsin deficiency. Methods By exploiting wave properties of x-rays for contrast formation, dark-field chest x-ray visualises the structural integrity of the alveoli, represented by a high signal over the lungs in the dark-field image. We investigated four patients with alpha1-antitrypsin deficiency with a novel dark-field x-ray prototype and simultaneous conventional chest x-ray. The extent of pulmonary function impairment was assessed by pulmonary function measurement and regional emphysema distribution was compared with CT in one patient. Results We show that dark-field chest x-ray visualises the extent of pulmonary emphysema displaying severity and regional differences. Areas with low dark-field signal correlate with emphysematous changes detected by computed tomography using a threshold of -950 Hounsfield units. The airway parameters obtained by whole-body plethysmography and single breath diffusing capacity of the lungs for carbon monoxide demonstrated typical changes of advanced emphysema. Conclusions Dark-field chest x-ray directly visualised the severity and regional distribution of pulmonary emphysema compared to conventional chest x-ray in patients with alpha1-antitrypsin deficiency. Due to the ultra-low radiation dose in comparison to computed tomography, dark-field chest x-ray could be beneficial for long-term follow-up in these patients.

Microalgae are major antioxidant producers and feed containing these substances is known to be beneficial. Microalgae cultivation is an alternative way to produce biodiesel and, after oil extraction, residual algal biomass (RAB) is obtained. The RAB was tested as an ingredient in fish feed production and its safety evaluation is important to prevent risks to fish health. This study aim was to evaluate, through biochemical and genetic biomarkers, the safety of RAB in catfish, Rhamdia quelen, feed. Acutodesmus obliquus microalgae RAB, cultivated in Chu medium, was used in feed formulation. A standard feed without RAB (0%) was produced, and three other feeds were enriched with RAB in 1, 2, and 3% proportion. Each feed kind was given to a 15 R. quelen fingerling group for 60 days. The evaluated biochemical biomarkers were superoxide dismutase (SOD) and catalase (CAT) activities, lipid peroxidation (LPO) in the liver, and acetilcolinesterase (AChE) activity in the brain and muscles. The genetic biomarkers analyzed were halo assay in erythrocytes, comet assay in erythrocytes, liver and brain, and piscine micronucleus. The SOD activity was increased in the 3% group; CAT activity and LPO levels were not different among the groups. In the comet assay, a significant decrease in DNA damage in erythrocytes (2 and 3%) and liver tissue (3%) was observed. In the brain, DNA damage was not observed. These results corroborate that as the RAB amount increased, the organisms showed a potential antioxidant effect, as the 3% RAB feed had the best results.

Context. —Gastrin-releasing peptide receptors (GRPRs) activate mitogen-activated protein kinase signaling pathway primarily through epidermal growth factor receptor activation and are under investigation as a molecular target because they are overexpressed in several solid tumors. Objective. —To determine GRPR expression in both non–small cell lung carcinoma and small cell lung carcinoma, comparing results with clinical stages and demographic data. Design. —We analyzed the immunohistochemical expression of GRPR in 200 non–small cell lung carcinoma and 38 small cell lung carcinoma archival cases from 2004 to 2008. Results. —Non–small cell lung carcinoma cases tended to be higher GRPR expressers at a rate of 62.5% (weak, moderate, and strong expression in 41.5%, 13.5%, and 7.5%, respectively), compared with 52.62% in small cell lung carcinoma cases (weak, moderate, and strong expression in 34.21%, 15.78%, and 2.63%, respectively; P = .30). In non–small cell lung carcinoma there was a trend for higher percentages of strong expression in adenocarcinoma cases (10%; P = .67), and in patients with advanced stages (III and IV; 9.43% and 6.9%; P = .01). Conclusions. —To the best of our knowledge, this is the first study to demonstrate GRPR tissue expression in a large population of patients with lung cancer. Although GRPR expression was similar in small cell and non–small cell carcinoma, the expression was more pronounced in an advanced-stage lung cancer, particularly in adenocarcinoma cases, and may represent a potential target for the development of new treatment approaches in this population.

Non-diphtheriae Corynebacterium species are usually considered as contaminants of clinical specimens due to their widely environmental distribution and colonization of the human skin and mucous membranes. However, these bacteria have been increasingly recognized as agents of life-threatening infections mainly in individuals in immunosuppressive conditions. These organisms have vast variation in morphology and biochemical reaction, characteristics that make the correct identification of Corynebacterium at the species level extremely difficult using conventional phenotypic methods. The precise identification of C. amycolatum requires approaches rarely available in conventional clinical microbiology laboratories, such as API Coryne system, 16s rRNA and rpoB gene sequencing. In this setting, MALDI-TOF, a quick, accurate, and relatively unexpansive molecular technique, arises as a cost-effective alternative for characterizing these agents. Here, a rare and lethal case of endocarditis caused by C. amycolatum is presented. This is the first case of infective endocarditis due to C. amycolatum reported in Brazil.