The turning point of COVID-19 severity is associated with a unique circulating neutrophil gene signature.

COVID-19 has a broad spectrum of clinical manifestations associated with the host immune response heterogeneity. Despite the advances in COVID-19 research, it is still crucial to seek a panel of molecular markers that enable accurate stratification of COVID-19 patients. Here, we performed a study that combined analysis of blood transcriptome, demographic data, clinical aspects and laboratory findings from 66 participants classified into different degrees of COVID-19 severity and healthy subjects. We identified a perturbation in blood-leukocyte transcriptional profile associated with COVID-19 aggravation, which was mainly related to processes that disfavoured lymphocyte activation and favoured neutrophil activation. This transcriptional profile stratified patients according to COVID-19 severity. Hence, it enabled identification of a turning point in transcriptional dynamics that distinguished disease outcomes and non-hospitalized from hospitalized moderate patients. Central genes of this unique neutrophil signature were S100A9, ANXA3, CEACAM6, VNN1, OLFM4, IL1R2, TCN1 and CD177. Our study indicates the molecular changes that are linked with the differing clinical aspects presented by humans when suffering from COVID-19, which involve neutrophil activation.

Acetylcholine, Fatty Acids, and Lipid Mediators Are Linked to COVID-19 Severity.

Lipid and cholinergic mediators are inflammatory regulators, but their role in the immunopathology of COVID-19 is still unclear. Here, we used human blood and tracheal aspirate (TA) to investigate whether acetylcholine (Ach), fatty acids (FAs), and their derived lipid mediators (LMs) are associated with COVID-19 severity. First, we analyzed the perturbation profile induced by SARS-CoV-2 infection in the transcriptional profile of genes related to the ACh and FA/LM pathways. Blood and TA were used for metabolomic and lipidomic analyses and for quantification of leukocytes, cytokines, and ACh. Differential expression and coexpression gene network data revealed a unique transcriptional profile associated with ACh and FA/LM production, release, and cellular signaling. Transcriptomic data were corroborated by laboratory findings: SARS-CoV-2 infection increased plasma and TA levels of arachidonic acid, 5-hydroxy-6E,8Z,11Z,14Z-eicosatetraenoic acid, 11-hydroxy-5Z,8Z,12E,14Z-eicosatetraenoic acid, and ACh. TA samples also exhibited high levels of PGE2, thromboxane B2, 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid, and 6-trans-leukotriene B4 Bioinformatics and experimental approaches demonstrated robust correlation between transcriptional profile in Ach and FA/LM pathways and parameters of severe COVID-19. As expected, the increased neutrophil-to-lymphocyte ratio, neutrophil counts, and cytokine levels (IL-6, IL-10, IL-1ß, and IL-8) correlated with worse clinical scores. Glucocorticoids protected severe and critical patients and correlated with reduced Ach levels in plasma and TA samples. We demonstrated that pulmonary and systemic hyperinflammation in severe COVID-19 are associated with high levels of Ach and FA/LM. Glucocorticoids favored the survival of patients with severe/critical disease, and this effect was associated with a reduction in ACh levels.

Cottonseed meal with enzyme complex for finishing pigs.

This work evaluated the digestibility of the experimental diets, the performance and the economic viability of pigs in the finishing phase fed with diets containing increasing levels of cottonseed meal (CM) with enzymatic complex. In the digestibility test, 35 barrows with an average weight of 54.54 ± 2.49 kg were used and for the performance and economic viability test, 30 pigs with an average weight of 55.47 ± 5.3 kg were used. In both trials, the animals were distributed in five diets: control diet (0), control diet plus the enzyme complex (0E), diet with 5% CM plus the enzyme complex (5E), diet with 10% CM plus the enzyme complex (10E), and a diet with 15% CM plus the enzyme complex (15E). The digestibility coefficients of dry matter (DM), organic matter (OM) and crude protein (CP) reduced with the inclusion of CM with enzyme complex in the diet of pigs in the finishing phase, in contrast to the ether extract (EE), which improved as the inclusion of CM increased. No statistical difference (P > 0.05) was found in the performance when all diets were compared; however, when comparing only the diets that contained the enzyme complex separately, a negative linear effect (P = 0.015) was observed for the feed conversion (FC) in period 2. The diets have no effects on periods 1 and 3. Diets 0, 5E, 10E, and 15E, presented better net income (P = 0.019) in March to May 2018 due to the different levels of inclusion of CM with enzyme complex; in the other evaluated items, there were no statistical differences (P > 0.05). The inclusion of CM and enzyme complex in the diets reduced the digestibility of the dietary components; however, it did not reflect on the performance and was not financially viable in the studied scenario.

IRMS as a tool to obtain the carbon turnover (δ13 C) in organs of weaned piglets fed glutamic acid and nucleotides.

Early weaning of piglets causes stress characterized by a decrease in feed intake followed by a decline in growth rates; thus, a fast recovery represents an essential step for proper growth of these animals. Considering that IRMS is a potential tool for non-destructive sampling and the fact that it provides time-integrated estimate of assimilated and not just ingested nutrients turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ13 C) in organs of weanling piglets. At day 0, three piglets were slaughtered (prior to diet switch), the remaining eighty-four piglets weaned at 21-day-old were randomly assigned in a complete block design with a 2 × 2 factorial arrangement of treatments (two Nu levels: 0 and 0.1% and two Glu levels: 0 and 1%), being three piglets per treatment slaughtered on trial days 3, 6, 9, 14, 21, 35 and 49. The samples were analysed by IRMS and adjusted to first-order equation by a non-linear regression analysis using NLIN of SAS, in order to establish exponential graphics. After that, the turnover data were submitted to analysis of variance using GLM of SAS. The turnover value (t95% ) verified for spleen was faster (p < 0.05) when glutamate was supplemented in diets. For pancreas and liver, the turnover rates were faster (p < 0.05) for the mixture of additives. However, for renal tissue, the turnover rate (t95% ) was greater (p < 0.05) for the free additive diet. The results obtained suggest that the mixture of additives was more efficient to develop the digestive tract at post-weaning phase, taking into account the functional importance of pancreas and liver for nutrients' digestion and processing.

Timing carbon turnover (δ13C) in weaned piglet's brain by IRMS.

Isotope-ratio mass spectrometry (IRMS) is a potential tool that provides time-integrated estimate of assimilated and not just ingested nutrients. This feature turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ13C) in the brain of weanling piglets. Eighty-seven piglets weaned at 21-day-old were used, being three piglets slaughtered at day-0 (prior to diet-switch) and, the remaining 84 randomly assigned in a complete block design with a 2 x 2 factorial arrangement of treatments (two nucleotide levels: 0% and 0.1% and, two glutamate levels: 0% and 1%). On trial days 3, 6, 9, 14, 21, 35 and 49, three piglets per treatment were also slaughtered. The samples were analyzed by IRMS and adjusted to the first-order equation by a nonlinear regression analysis using NLIN procedure of SAS, in order to obtain the exponential graphics. The carbon turnover (t95%) verified for cerebral tissue was faster (P<0.05) for diet containing glutamate in comparison to other diets, supporting the fact that glutamate contributed to develop the piglets' brain, due to the fastest incorporation of 13C-atoms in this tissue at post-weaning phase, despite the energy deficit experienced by them.

Evaluation of growth performance and gastro-intestinal parameters on the response of weaned piglets to dietary organic acids

ABSTRACT Two experiments (E) were carried out to evaluate the effects of fumaric acid and an acidifier blend [composed by calcium formate, calcium lactate and medium-chain fatty acids (capric and caprylic)] in piglet diets containing colistin (40 ppm) or halquinol (120 ppm) on performance, diarrhea incidence (E1), organs relative weight, pH values, intestinal morphometry and microbiota (E2). In E1, 192 and E2, 24 piglets weaned at 21-day-old were randomly assigned to blocks with 2x2 factorial arrangement of treatments [absence or presence of fumaric acid x absence or presence of acidifier blend], six replicates of eight (E1) and one piglet per pen (E2). For E1, the treatments were control (CD): no acidifier product + 40 ppm of colistin, FA: fumaric acid in absence of acidifier blend, AB: acidifier blend in absence of fumaric acid and, AF+AB: presence of fumaric acid and acidifier blend. For E2, the pre-starter I diet were used and the same treatments as E1 evaluated. No treatment effects (P>0.05) were observed on performance, diarrhea incidence (E1), gut pH values and duodenum morphometry of piglets (E2). However, the addition of AB increased (P<0.05) large intestine relative weight and, FA addition decreased (P<0.05) pancreas relative weight, jejunum villi height and, total coliform and E. coli counts in cecum. The inclusion of FA and AB in diets containing colistin or halquinol did not improve performance, although FA exerted an inhibitory effect on cecum microbiota.

Evaluation of growth performance and gastro-intestinal parameters on the response of weaned piglets to dietary organic acids.

Two experiments (E) were carried out to evaluate the effects of fumaric acid and an acidifier blend [composed by calcium formate, calcium lactate and medium-chain fatty acids (capric and caprylic)] in piglet diets containing colistin (40 ppm) or halquinol (120 ppm) on performance, diarrhea incidence (E1), organs relative weight, pH values, intestinal morphometry and microbiota (E2). In E1, 192 and E2, 24 piglets weaned at 21-day-old were randomly assigned to blocks with 2x2 factorial arrangement of treatments [absence or presence of fumaric acid x absence or presence of acidifier blend], six replicates of eight (E1) and one piglet per pen (E2). For E1, the treatments were control (CD): no acidifier product + 40 ppm of colistin, FA: fumaric acid in absence of acidifier blend, AB: acidifier blend in absence of fumaric acid and, AF+AB: presence of fumaric acid and acidifier blend. For E2, the pre-starter I diet were used and the same treatments as E1 evaluated. No treatment effects (P>0.05) were observed on performance, diarrhea incidence (E1), gut pH values and duodenum morphometry of piglets (E2). However, the addition of AB increased (P<0.05) large intestine relative weight and, FA addition decreased (P<0.05) pancreas relative weight, jejunum villi height and, total coliform and E. coli counts in cecum. The inclusion of FA and AB in diets containing colistin or halquinol did not improve performance, although FA exerted an inhibitory effect on cecum microbiota.

Dietary glutamine, glutamic acid and nucleotide supplementation accelerate carbon turnover (δ13C) on stomach of weaned piglets.

The use of stable isotope analysis as a tool for characterization of carbon turnover (δ13C) in piglet's tissues by tracing its feeding system has drawn attention. Thus, this study aimed at evaluating the influence of dietary glutamine, glutamic acid and nucleotides supplementation on carbon turnover in fundic-stomach region of weaned piglets at an average age of 21 days. The diets consisted of additive-free diet - control (C); 1% glutamine (G); 1% glutamic acid (GA) and 1% nucleotides (Nu). At weaning day (day 0: baseline), 3 piglets were slaughtered to quantify the δ13C of stomach. The remaining 120 piglets were blocked by weight and sex, randomly assigned to pens with 3 piglets slaughtered per treatment at days 1, 2, 4, 5, 7, 9, 13, 20, 27 and 49 after weaning in order to verify the fundic-stomach isotopic composition by treatments. Samples were analyzed in terms of 13C/12C ratio by mass spectrometry and converted to relative isotopic enrichment values (δ13C ‰) used to plot the first order exponential curves over time using OriginPro 8.0 software. The inclusion of glutamine, glutamate and nucleotides in piglet's diets has accelerated the carbon turnover in stomach during the post-weaning period, demonstrating also that glutamate has guaranteed fastest 13C incorporation rate on fundic-stomach region and pH-lowering. Besides that, stable isotopes technique (δ13C) has proved to be an important methodology to determine the time-scales at which piglets shift among diets with different isotopic values, characterizing the trophic effects of additives and the phenotypic flexibility of stomach.

Effect of Glutamine, Glutamic Acid and Nucleotides on the Turnover of Carbon (δ(13)C) in Organs of Weaned Piglets.

Morphological and physiological alterations occur in the digestive system of weanling piglets, compromising the performance in subsequent phases. This experiment aimed at verifying the influence of glutamine, glutamate and nucleotides on the carbon turnover in the pancreas and liver of piglets weaned at 21 days of age. Four diets were evaluated: glutamine, glutamic acid or nucleotides-free diet (CD); containing 1% glutamine (GD); containing 1% glutamic acid (GAD) and containing 1% nucleotides (ND). One hundred and twenty-three piglets were utilized with three pigs slaughtered at day zero (weaning day) and three at each one of the experimental days (1, 2, 4, 5, 7, 9, 13, 20, 27, and 49 post-weaning), in order to collect organ samples, which were analyzed for the δ(13)C isotopic composition and compared by means of time. No differences were found (p>0.05) among treatments for the turnover of the (13)C in the pancreas (T50% = 13.91, 14.37, 11.07, and 9.34 days; T95% = 46.22, 47.73, 36.79, and 31.04 days for CD, GD, GAD, and ND, respectively). In the liver, the ND presented accelerated values of carbon turnover (T50% = 7.36 and T95% = 24.47 days) in relation to the values obtained for the GD (T50% = 10.15 and T95% = 33.74 days). However, the values obtained for the CD (T50% = 9.12 and T95% = 30.31 days) and GAD (T50% = 7.83 and T95% = 26.03 days) had no differences (p>0.05) among other diets. The technique of (13)C isotopic dilution demonstrated trophic action of nucleotides in the liver.

Nutritional evaluation of integral cassava root silages for growing pigs.

The experiment aimed at determining the nutritional value of integral cassava root silages with yogurt as inoculant or wastewater (manipueira) by liquid addition ensiling process. Eighteen crossbred piglets (Large White × Landrace), castrated males with an initial average weight of 50 kg were allocated in metabolism cages throughout 11-day trial duration (6 days for animal's adaptation to the cages and to the experimental diets, and 5 days of urine and faeces collection). The experimental design was a randomized block arrangement with 3 treatments and 6 replicates: basal diet (100%); mixture composed of 75% basal diet and 25% integral cassava root silage with yogurt as inoculant; a mixture composed of 75% of basal diet and 25% integral cassava root silage with wastewater. The silages with wastewater and yogurt presented the following values of apparent digestibility of dry matter 89.96% and 90.01%, apparent digestibility of crude protein of 60.67% and 66.43%, apparent digestibility of gross energy of 90.43% and 91.48%, gross energy metabolizability coefficients of 87.88% and 88.93%, digestible energy values of 3,705 and 3,783 kcal/kg on a dry matter (DM) basis, and metabolizable energy values of 3,600 and 3,676 kcal/kg DM, respectively. The results have demonstrated that integral cassava root silages with wastewater or yogurt have a high nutritional value and can be used as an alternative energy source in growing pig's diets.

Effects of ß-(1→3,1→6)-D-glucan and density of diets on the blood profiles of immunologically challenged weaned piglets.

An experiment was conducted to evaluate the effects of two levels of the ß-(1→3,1→6)-D-glucan (0 and 500 ppm) from yeast Saccharomyces cerevisiae and two levels of energy (3300 and 3450 kcal ME kg(-1)) on the hematological, immunological and, biochemical profiles of thirty-six 21-days-old weaned piglets, challenged with 150 µg kg(-1) of BW lipopolysaccharide (LPS) from Escherichia coli serotype 055:B5. The experimental design was a randomized complete block design in a 2×2 factorial arrangement with nine replicates per treatment and, one animal per experimental unit. The data were analyzed in accordance with the multivariate analysis procedure of SAS and, the treatment means of parametric and non-parametric data were compared by Bonferroni's test (P<0.05) and, by Dunn's test (P<0.05), respectively. The data of the blood profiles of alanine aminotransferase, alkaline phosphatase and, creatinine showed that LPS did not cause kidney or liver damage in the animals. The addition of beta-glucan in the diets did not prove the robustness of its effect and biological relevance when provided with low nutrient-density. However, its addition combined with the high-nutrient-density diets showed less marked hypoglobulinemia in piglets, which may have contributed to the decreasing of the synthesis of inflammatory mediators.

Inorganic and organic trace mineral supplementation in weanling pig diets.

A study was conducted to evaluate the effects of dietary inorganic and organic trace minerals in two levels of supplementation regarding performance, diarrhea occurrence, hematological parameters, fecal mineral excretion and mineral retention in metacarpals and liver of weanling pigs. Seventy piglets weaned at 21 days of age with an average initial body weight of 6.70 ± 0.38 kg were allotted in five treatments: control diet (no added trace mineral premix); 50% ITMP (control diet with inorganic trace mineral premix supplying only 50% of trace mineral requirements); 50% OTMP (control diet with organic trace mineral premix supplying only 50% of trace mineral requirements); 100% ITMP (control diet with inorganic trace mineral premix supplying 100% of trace mineral requirements); and 100% OTMP (control diet with organic trace mineral premix supplying 100% of trace mineral requirements). Feed intake and daily weight gain were not affected by treatments, however, piglets supplemented by trace minerals presented better gain:feed ratio. No differences were observed at calcium, phosphorus, potassium, magnesium, sodium and sulfur excreted in feces per kilogram of feed intake. Treatments did not affect calcium, phosphorus, magnesium, sulfur and iron content in metacarpals. Trace mineral supplementation, regardless of level and source, improved the performance of piglets.

Aerococcus viridans como agente etiológico de endocardite em valva nativa / Aerococcus viridans as etiological agent in native valve endocarditis

Aerococcus viridans é um patógeno humano incomum e endocardite por este agente na literatura é rara. Relata-se o caso de paciente feminina, 56 anos,hospitalizada devido a quadro de picos febris que, após investigação, foi diagnosticada através dos critérios clínicos de Duke, com endocardite em valva nativa. Foi evidenciado através das hemoculturas o crescimento de A. viridans, sendo o primeiro casoreportado no Brasil. Há relatos desse agente também em infecções urinárias, bacteremia, meningite e artrite séptica. No presente caso, a paciente teve melhorresposta clínica após o início da ampicilina, confirmando alguns artigos que orientam o tratamento com penicilinas.

Aerococcus viridans is an uncommon human pathogen and reports of endocarditis caused by this agent arerare in the literature. This clinical case study describes a 56 year-old female patient admitted to hospital forspiking fevers. After investigation, this was diagnosed as native valve endocarditis, based on the Duke clinicalcriteria and confirmed through the growth of A. viridans in blood cultures, making this the first reported case in Brazil. There are also reports of thisagent in urinary tract infections, bacteremia, meningitis and septic arthritis. In this case, the patient had a better clinical response after starting on ampicillin, confirmed by some articles that recommend treatment withpenicillins.