Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens

Foot-and-mouth disease (FMD), caused by the FMD virus (FMDV), is controlled by vaccine policy in many countries. For vaccine potency, the content of intact virus particles (146S antigens) is critical, and the sucrose density gradient (SDG) fractionation is the gold standard for the quantification of 146S antigens. However, this method has several drawbacks. Although size-exclusion high-performance liquid chromatography (SE-HPLC) was introduced to replace the classic method, its application is generally confined to purified samples owing to the interfering signals. Therefore, we aimed to develop optimal pretreatment methods for SE-HPLC quantification in less purified samples. Crude virus infection supernatant (CVIS) and semi-purified samples with PEG precipitation (PEG-P) were used. Chloroform pretreatment was essential to remove a high level of non-specific signals in CVIS, whereas it caused loss of 146S antigens without the distinctive removal of non-specific signals in PEG-P. Benzonase pretreatment was required to improve the resolution of the target peak in the chromatogram for both CVIS and PEG-P. Through spiking tests with pure 146S antigens, it was verified that the combined pretreatment with chloroform and benzonase was optimal for the CVIS, while the sole pretreatment of benzonase was beneficial for PEG-P.

Use of Ionic Liquid Pretreated and Fermented Sugarcane Bagasse as an Adsorbent for Congo Red Removal

A large amount of industrial wastewater containing pollutants including toxic dyes needs to be processed prior to its discharge into the environment. Biological materials such as sugarcane bagasse (SB) have been reported for their role as adsorbents to remove the dyes from water. In this study, the residue SB after fermentation was utilized for the dye removal. A combined pretreatment of NaOH and methyltrioctylammonium chloride was given to SB for lignin removal, and the pretreated SB was utilized for cellulase production from Bacillus aestuarii UE25. The strain produced 118 IU mL−1 of endoglucanse and 70 IU mL−1 of β-glucosidase. Scanning electron microscopy and FTIR spectra showed lignin and cellulose removal in fermented SB. This residue was utilized for the adsorption of an azo dye, congo red (CR). The thermodynamic, isotherm and kinetics studies for the adsorption of CR revealed distinct adsorption features of SB. Untreated SB followed Langmuir isotherm, whereas pretreated SB and fermented SB obeyed the Freundlich isotherm model. The pseudo-second-order model fitted well for the studied adsorbents. The results of thermodynamic studies revealed spontaneous adsorption with negative standard free energy values. Untreated SB showed a 90.36% removal tendency at 303.15 K temperature, whereas the adsorbents comprised of pretreated and fermented SB removed about 98.35% and 97.70%, respectively. The study provided a strategy to utilize SB for cellulase production and its use as an adsorbent for toxic dyes removal.

Alkaline electro-hydrolysis pretreatment of rice straw for enhanced biogas production under ambient temperature

Biogas production can supplement the renewable energy target of the world. For this abundantly available agricultural waste like wheat and rice straw can be used. Biogas generation using this waste will curb the stubble burning incidences, reduce greenhouse gases, enhance farmer’s income, and strengthen the energy security of many countries. The recalcitrant nature of rice straw is a barrier to its hydrolysis, which is a prerequisite of the anaerobic digestion (AD) process. Alkaline, electro-hydrolysis, and a combination of both pretreatment (alkaline electrohydrolysis) methods are undertaken in the present study. Batch reactors at ambient temperature were used for AD of rice straw at different inoculums to substrate ratios (ISR) of 0.5, 0.75, and 1.0 to observe the effect on biogas/methane yield. Among these, a higher amount of biogas was obtained for ISR of 0.5 in all sets of experiments. The combined pretreatment method yielded biogas of 315.9 mL/gVS (equivalent to methane of 167.4 mL/gVS). For pretreat-ed rice straw by electro-hydrolysis, alkaline, and combined (alkaline and electro-hydrolysis) respectively, the methane yields were 7.03, 18.13, and 49.82% higher than untreated rice straw. The biogas had approximately 53% of methane content. The use of rice straw for biogas production may prove a viable alternative for clean and sustainable energy. In the studies, a first-order kinetic model is found to fit better the experimental results.

Pretreatment with Coenzyme Q10 Combined with Aescin Protects against Sepsis-Induced Acute Lung Injury

Sepsis-associated acute lung injury (ALI) is a critical condition characterized by severe inflammatory response and mitochondrial dysfunction. Coenzyme Q10 (CoQ10) and aescin (AES) are well-known for their anti-inflammatory activities. However, their effects on lipopolysaccharide (LPS)-induced lung injury have not been explored yet. Here, we asked whether combined pretreatment with CoQ10 and AES synergistically prevents LPS-induced lung injury. Fifty male rats were randomized into 5 groups: (1) control; (2) LPS-treated, rats received a single i.p. injection of LPS (8 mg/kg); (3) CoQ10-pretreated, (4) AES-pretreated, or (5) combined-pretreated; animals received CoQ10 (100 mg/kg), AES (5 mg/kg), or both orally for 7 days before LPS injection. Combined CoQ10 and AES pretreatment significantly reduced lung injury markers; 52.42% reduction in serum C-reactive protein (CRP), 53.69% in alkaline phosphatase (ALKP) and 60.26% in lactate dehydrogenase (LDH) activities versus 44.58, 37.38, and 48.6% in CoQ10 and 33.81, 34.43, and 39.29% in AES-pretreated groups, respectively. Meanwhile, combination therapy significantly reduced interleukin (IL)-1β and tumor necrosis factor (TNF)-α expressions compared to monotherapy (<i>p</i> &#x3c; 0.05). Additionally, combination therapy prevented LPS-induced histological and mitochondrial abnormalities greater than separate drugs. Western blotting indicated that combination therapy significantly suppressed nucleotide-binding oligomerization domain (NOD)-like receptors-3 (NLRP-3) inflammasome compared to separate drugs (<i>p</i> &#x3c; 0.05). Further, combination therapy significantly decreased the expression of signaling cascades, p38 mitogen-activated protein kinases (p38 MAPK), nuclear factor kappa B (NF-κB)-p65, and extracellular-regulated kinases 1/2 (ERK1/2) versus monotherapy (<i>p</i> &#x3c; 0.05). Interestingly, combined pretreatment significantly downregulated high mobility group box-1 (HMGB1) by 72.93%, and toll-like receptor 4 (TLR4) by −0.93-fold versus 61.92%, −0.83-fold in CoQ10 and 38.67%, −0.70-fold in AES pretreatment, respectively. Our results showed for the first time that the enhanced anti-inflammatory effect of combined CoQ10 and AES pretreatment prevented LPS-induced ALI via suppression of NLRP-3 inflammasome through regulation of HMGB1/TLR4 signaling pathway and mitochondrial stabilization.

Xylanase pretreatment of mechanically destructured chips of Eucalyptus tereticornis and its effect on kraft pulping

Mechanical pulping of raw wood material is a highly energy intensive and pollution generating step in the papermaking process. This study focused on combined mechanical and xylanase treatment prior to the kraft pulping of E. tereticornis. A screened pulp yield of 49.1% (on oven-dry wood basis) with a Kappa number of 24.9 was obtained at the optimum cooking temperature of 160 °C without any pretreatment of the wood chips. After mechanical treatment (destructuring), a slightly higher screened pulp yield (49.4%) was obtained with a Kappa number of 24.2 at the cooking temperature of 145 °C with the same active alkali charge (15%). The optimum cooking temperature was further reduced to 140 °C for the destructured xylanase-treated wood chips. The xylanase treatment resulted in a 2% reduction in screened pulp yield due to hydrolysis of xylan. However, the Kappa number was reduced to 18.2 after xylanase pretreatment of the mechanically destructured wood chips. The combined pretreatment (destructured and xylanase treatment) of wood chips resulted in a reduction in cooking temperature by 20°C compared to untreated wood chips. Such a reduction in cooking temperature can effectively reduce steam consumption. The combined pretreatment improved the pulp brightness by 2.0 (ISO points) and physical strength properties, which included the tensile index, tear index, and burst index by 11.06%, 21.72%, and 21.79%, respectively, compared to the control.

Evaluation Of Mechanical, Irradiation And Chemical Pretreatment Lignocelulosic Substrate For Enhanced Methane Production

Lignocellulosic substrate is a resource that contains a locked energy reserve that is normally lost during anaerobic digestion. Lignocellulosic substrate is one of the most abundant sources of organic matter available and yet its energy recovery has much room for improvement. Lignocellulosic substrate has cellular properties that are deemed extremely difficult to degrade due to complexity which is why this energy reserve is never unlocked during anaerobic digestion. There are several successful pretreatment methods that are used to degrade this lignocellulosic substrate and unlock this energy reserve. This paper will focus on the methods that include mechanical, irradiation, chemical and combined pretreatment processes. Analysis is conducted on all the studies that are obtained to compare the successes of the different types of pretreatment processes used. Each of the different listed pretreatment processes have different energy requirements, treatment times, and solvent requirement and are acting to enhancing methane production. The improvement in methane production varies from process to process and study to study creating a need to compile all of this valuable data into this research report. This will help future researchers in navigating the available studies of pretreatment of lignocellulosic substrate for improving methane production.