Accelerating strain phenotyping with desorption electrospray ionization-imaging mass spectrometry and untargeted analysis of intact microbial colonies

Reading and writing DNA were once the rate-limiting step in synthetic biology workflows. This has been replaced by the search for the optimal target sequences to produce systems with desired properties. Directed evolution and screening mutant libraries are proven technologies for isolating strains with enhanced performance whenever specialized assays are available for rapidly detecting a phenotype of interest. Armed with technologies such as CRISPR-Cas9, these experiments are capable of generating libraries of up to 1010 genetic variants. At a rate of 102 samples per day, standard analytical methods for assessing metabolic phenotypes represent a major bottleneck to modern synthetic biology workflows. To address this issue, we have developed a desorption electrospray ionization–imaging mass spectrometry screening assay that directly samples microorganisms. This technology increases the throughput of metabolic measurements by reducing sample preparation and analyzing organisms in a multiplexed fashion. To further accelerate synthetic biology workflows, we utilized untargeted acquisitions and unsupervised analytics to assess multiple targets for future engineering strategies within a single acquisition. We demonstrate the utility of the developed method using Escherichia coli strains engineered to overproduce free fatty acids. We determined discrete metabolic phenotypes associated with each strain, which include the primary fatty acid product, secondary products, and additional metabolites outside the engineered product pathway. Furthermore, we measured changes in amino acid levels and membrane lipid composition, which affect cell viability. In sum, we present an analytical method to accelerate synthetic biology workflows through rapid, untargeted, and multiplexed metabolomic analyses.

Comparative analysis production of fruit and sunti acid using the Hayami method

Agroindustry sector has a role important to improve the economic society level of Aceh, Indonesian. The opportunities in business to develop agroindustry from Averrhoa bilimbi L. was a relative major potential included the fruit candy and sunti acid. This study aims to calculated added value of fruit candy and sunti acid using the hayami method. The result was obtained primary and secondary data. The primary data were obtained directly results of interviews with business actors using questionnaires. Furthermore, the secondary data were obtained from literature review studies. The data was collected using the observation, interviews, and documentation. The average added value obtained by fruit candy of each kilogram was found the output value in Rp.150,000 Whereas, the sunti acid was a lower value in Rp. 65,000 This means that a kilogram was used can provide added value of Rp. 11,250 from fruit candy and Rp. 15,625 from sunti acid. The added value ratio was used to quantify the advancement achieved for productivity from A. bilimbi in agroindustry to involve the additional value from local products. Then, the highest profit until 60% in sunti acid product, and 44.44% in fruit candy. The retribution for the owner of the production factor was found the value the entrepreneur profit to 9.76% (Rp. 51,250/kg) for fruit candy and 26.37% (Rp.35,625/kg) for sunti acid. The descriptive data analysis method was expected to suggest the development of agroindustry based on the local commodity product, mainly the fruit candy and sunti acid to supported the food security.

Milk quality and its suitability for technological processing in cows with metritis

It has been established that the development of metritis in cows after providing obstetric aid as a result of abortion, eversion of the uterus or retention of the placenta is accompanied by an increased microbial and fungal background of the uterus. Without obstetric aid during delivery, only from 5… 9 days after birth, 35.37% of cows had genitals contaminated with various pathogenic microflora. Already on the third day of puerperia, 14 species of bacteria were isolated from cows that were assisted in delivery, which in 74.5% of cases were contaminated with pathogenic microflora: S. aureus (in 15.5% of cases), E. coli (37%), K. pneumonia (12%), and S. pyogenes (10% of cases). The results of mycological studies revealed that A. fumigatus, C. albicans and C. crusei were isolated from cows after obstetrics. It was found out that the content of somatic cells (SC) r = 0.63, the activity of muramidase (AM) r = 0.84, lactoperoxidase (LPO) r = 0.65 and lactoferrin (LF) r = 0.66 change with a high degree of correlation. Milk from cows with metritis showed 2 times higher total bacterial contamination than milk from clinically healthy animals. Milk from sick cows has a reduced number of lactic acid organisms after the first day of storage. At the same time, acid formation occurred faster by 5.0–15.0% than that in control samples of milk prepared for production of lactic acid products. The acidity in milk fermented with Lactobacillus bulgarus was 12.0-13.3% higher than that in the control sourdough samples, and the cell viability of the symbiotic combination was an order of magnitude lower (2.5×106 versus 2.5×107) compared to the control samples of the lactic acid product.

Impact of lactic acid product on quality indices of raw meat for the smoked sausages production

Conducted research pertains to technologies of animal products processing and the development of biotechnological methods for stabilizing microbial and enzymatic biological leaven for meat.Materials and Methods. The studies were conducted in the conditions of the Research Institute of Food and Animal Processing Technologies and Laboratory of Microbiological Research Methods, Department of Microbiology of Bila Tserkva National Agrarian University. Moisture retaining capacity and bacterial contamination of beef and pork were determined.Results. The effect of processing raw meat with lactic acid product – yogurt on the moisture retaining capacity was experimentally established. With the highest titratable acidity of yoghurt of 100-110 °T, the moisture retaining capacity of pork and beef was inferior to control on 2.13 and 1.29 % and amounted to 51.88 and 62.73 %. Under a titratable acidity of lactic acid beverage of 100-110 °T with total number of lactic acid bacteria on the surface of raw meat, it was 8.1×108 and 8.5×108, which prevailed samples with acidity level of 60-70 °T on 24.5 and 24.3 %.Conclusions. The results of studies have revealed the effect of processing raw meat by yogurt with different titratable acidity on moisture retaining capacity and bacterial contamination.

Catheter-induced anaphylaxis and determination of the causative catheter in a patient undergoing neuroendovascular surgery: a case report

Background Anaphylaxis caused by a catheter itself used for endovascular surgery is rare, and a method for detection of a causative catheter has not been established. We report a case of catheter-induced anaphylaxis in which the causative catheter was successfully detected. Case presentation A 47-year-old male underwent neuroendovascular surgery. During surgery, blood pressure suddenly dropped and the level of tryptase indicated the occurrence of anaphylaxis. There were 24 candidate agents for the cause of anaphylaxis including 8 catheters. We performed the basophil activation test by directly mixing the catheter with blood. One catheter coated with a hyaluronic acid product showed a positive reaction, and we confirmed the result by a modified skin test using an elution solution of the catheter. Later, we successfully completed the neuroendovascular surgery without the catheter. Conclusions The methods used in this case can be useful for the detection of the causative agent in catheter-induced anaphylaxis.

New insights on the UV/TiO2 photocatalytic treatment of thiomersal and its 2-sulfobenzoic acid product

Background: Thiomersal (TM), a complex between 2-mercaptobenzoic acid (2-MBA) and ethylmercury (C2H5Hg+), is an antimicrobial preservative used in immunological, ophthalmic, cosmetic products, and vaccines. Objective: TM has been treated by UV/TiO2 photocatalysis in the presence or absence of oxygen at acidic pH. C2H5Hg+, 2-MBA, and 2-sulfobenzoic acid (2-SBA) were found as products. A 2-SBA photocatalytic treatment was undertaken to study sulfur evolution. Methods: Photocatalytic runs were performed using a UVA lamp (λmax = 352 nm), open to the air or under N2. A suspension of the corresponding TM or 2-SBA salt and TiO2 was prepared, and pH was adjusted. Suspensions were stirred in the dark for 30 min and then irradiated. TM, 2-MBA, 2-SBA, and C2H5Hg+ were quantified by HPLC, sulfur by TXRF, and the deposits on the photocatalyst were analyzed by chemical reactions. The mineralization degree was followed by TOC. Sulfate was determined using BaCl2 at 580 nm. Results: Photocatalytic destruction of TM and total C2H5Hg+ was complete under N2 and air, but TM degradation was much faster in air. The evolution of TM and the products followed a pseudo-first-order kinetics. Conclusion: TiO2-photocatalytic degradation is a suitable technique for the treatment of TM and its degradation products. In contrast to other organomercurial compounds, TM degradation is faster in the presence of O2, indicating that the oxidative mechanism is the preferred pathway. A significant TM mineralization (> 60%, NPOC and total S) was obtained. TM was more easily degraded than 2-SBA. Sulfate was the final product.

Sodium buffered formic acid concentration and feed pH is stable over a 3-month period

Promoting feed hygiene with organic acids is an effective method to prevent foodborne illnesses from bacterial infection. The stability and acidification of mash and pelleted feed with sodium buffered formic acid was investigated. The acid product was incorporated to reach total formate inclusion levels of 0, 6, or 12 g/kg for swine nursery feed; 0, 4, or 9 g/kg for swine finishing feed; and 0, 3, or 6 g/kg for broiler grower feed. Samples were analyzed for total formate and pH on d 4, 32, 60, or 88 post-manufacturing. The concentration of formate remained stable across an 88-d period (P < 0.01). Treatment with the formic acid product decreased feed pH with increasing inclusion levels (all P < 0.01). Within each inclusion level of acid and across time, pH tended to increase in pelleted feeds and decrease in mash feeds (all P < 0.01); however, these changes were small (0.1 units pH). These data suggest that sodium buffered formic acid can be applied to both mash and pelleted feed to provide continuous acidification over a 3-month period.