Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The current work was carried out to investigate serine enantiomers in bacterial cells. The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive, rod shape, large, irregular, off-white-coloured, and synthesized endospores. A1C1 showed positive results (within 14±2h of incubation) for indole production, lactose fermentation, and protease (0.9 mm, clear zone). The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively within 12h of incubation. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells. Given results demonstrate the industrial significance and implication of the isolated strain for the synthesis of commercially valuable products.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97 ± 0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919 ± 0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85–90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150 ± 0.5 RPM, and 7 ± 0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Probiotic Bacteria and Their Cell Walls Induce Th1-Type Immunity Against Salmonella Typhimurium Challenge

Probiotics have been associated with a variety of health benefits. They can act as adjuvant to enhance specific immune response. Bacterial cell wall (CW) molecules are key structures that interact with host receptors promoting probiotic effects. The adjuvant capacity underlying this sub-cellular fraction purified from Lactobacillus casei CRL431 and L. paracasei CNCMI-1518 remains to be characterized. We interrogated the molecular and cellular events after oral feeding with probiotic-derived CW in addition to heat-inactivated Salmonella Typhimurium and their subsequent protective capacity against S. Typhimurium challenge. Intact probiotic bacteria were orally administered for comparison. We find that previous oral feeding with probiotics or their sub-cellular fraction reduce bacterial burden in spleen and liver after Salmonella challenge. Antibody responses after pathogen challenge were negligible, characterized by not major changes in the antibody-mediated phagocytic activity, and in the levels of total and Salmonella-specific intestinal sIgA and serum IgG, respectively. Conversely, the beneficial effect of probiotic-derived CW after S. Typhimurium challenge were ascribed to a Th1-type cell-mediated immunity which was characterized by augmentation of the delayed-type hypersensitivity response. The cell-mediated immunity associated with the oral feeding with probiotic-derived CW was accompanied with a Th1-cell polarizing cytokines, distinguished by increase IFN-γ/IL-4 ratio. Similar results were observed with the intact probiotics. Our study identified molecular events associated with the oral administration of sub-cellular structures derived from probiotics and their adjuvant capacity to exert immune modulatory function.

Viable circulating endothelial cells and their progenitors are increased in Covid-19 patients

During the course of Covid-19, the disease caused by the new Coronavirus SARS-CoV-2, thrombotic phenomena and/or diffuse vascular damage are frequent, and viral elements have been observed within endothelial cells. Circulating endothelial cells (CECs) and their progenitors (CEPs) are increased in cardiovascular, thrombotic, infectious and cancer diseases. Using a validated flow cytometry procedure, we found that viable CEPs/mL were significantly increased in Covid-19 patients compared to healthy controls. This increase was observed in patients with mild symptoms and not further augmented in patients with severe symptoms. In patients who recovered, CEPs decreased, but were in a range still significantly higher than normal controls. Regarding mature CECs, in Covid-19 patients their absolute number was similar to those observed in healthy controls, but the viable/apoptotic CEC ratio was significantly different. Both mild and severe Covid-19 patients had significantly more viable CECs compared to healthy controls. Patients who recovered had significantly less CECs/mL when compared to controls as well as to mild and severe Covid-19 patients. A positive correlation was found between the copies of SARS-CoV-2 RNA in the cellular fraction and apoptotic CEPs/mL in severe Covid-19 patients. These findings suggest that CECs and CEPs might be investigated as candidate biomarkers of endothelial damage in Covid-19 patients.

Cellular fraction analysis of pericardial effusion helps the diagnosis of eosinophilic myocarditis

Eosinophilic myocarditis is rare in children, and consequently, it is difficult to diagnose eosinophilic myocarditis rapidly. We report the clinical course of acute eosinophilic myocarditis with pericarditis in two adolescent boys and their associated electrocardiograms. The two patients, 13- and 14-year-old boys, developed cardiomegaly and chest pain with vomiting. On examination by two-dimensional echocardiography, thickening of the ventricular septum and a pericardial effusion were detected. The eosinophil count had increased by the pericardial effusion. Acute eosinophilic myocarditis often complicates a moderate to severe pericardial effusion owing to acute pericarditis. A cellular fraction analysis of the pericardial effusion is easy and useful for the diagnosis of eosinophilic myocarditis. Some serial changes in the electrocardiogram occur during each stage of acute eosinophilic myocarditis. They are induced by eosinophilic granules, which are capable of inducing tissue damage and dysfunction, and those changes in the electrocardiogram resemble the changes after an acute myocardial infarction. It is important to know the characteristics of eosinophilic myocarditis in order to prevent lethal complications.