We address the targeted destruction of Karenia brevis using the algaecide calcium peroxide, in tandem with the flocculation and sinking of the species. The specific aspect of the approach is the incorporation of the algaecide within the floc to rapidly kill K. brevis, thus minimizing escape of cells from the floc and reentry to the water column. CaO₂ gradually produces H₂O₂, which diffuses through cell membranes and induces oxidative stress, leading to cell death via excessive reactive oxygen species (ROS) formation. The effect of varying doses of calcium peroxide on K. brevis cells was measured with pulse amplitude modulated fluorometry and indicated that doses as low as 30 mg/L when integrated into flocs are effective in suppressing photosynthesis. Cell viability assays also indicate that such low levels are sufficient to cause cell death in a 3-6 hour time period. Thus, the proposed technology involving the incorporation of calcium peroxide in a cationic flocculating agent (polyaluminum chloride, PAC) leads to an inexpensive and scalable technology to mitigate harmful algal blooms of K. brevis.

This study aimed to investigate the immunogenicity of the hepatitis B virus (HBV) vaccine by applying a normal and high-dose hepatitis B virus vaccination program in the mice modeling of non-alcoholic fatty liver disease (NAFLD). NAFLD was induced in mouse livers via diet. At the 10-week mark, both groups were divided into 3 subgroups. While the standard dose vaccination program was applied on days 0, 7, and 21, two high-dose programs were applied: one was applied on days 0 and 7, and the other was applied on days 0, 7, and 21. All mice were euthanized. Blood samples from anti-HB titers; T follicular helper, T follicular regulatory, CD27+, and CD38+ cells; and the liver, spleen, and thymus were taken for histopathologic evaluation. NAFLD subgroups receiving high doses showed higher hepatocyte ballooning scores than normal-dose subgroup. There were differences in CD27+ and CD27+CD38+ cells in animals fed on different diets, without any differences or interactions in terms of vaccine protocols. In the NAFLD group, a negative correlation was observed between anti-HB titers and T helper and CD27+ cells, while a positive correlation was observed with CD38+ cells. NAFLD induced changes in immune parameters in mice, but there was no difference in vaccine efficacy among the applied vaccine protocols. Based on this study\'s results, high-dose vaccination protocols are not recommended in cases of NAFLD, as they do not enhance efficacy and may lead to increased liver damage.

We compared the immunogenicity of recombinant S. pneumoniae pneumolysin (rPly) when administered with and without Al(OH)3 adjuvant, and evaluated the protective properties of recombinant protein in the active defense experiment. It was shown that double immunization with rPly+Al(OH)3 increases the levels of IgG antibodies in comparison with the control (p<0.01), while triple immunization results in a more significant increase in IgG antibody levels (p<0.001). Double immunization with rPly without Al(OH)3 does not induce a significant increase in antibody levels in comparison with the control, while triple immunization results in a slight but significant increase in antibody levels (p<0.05). The active defense test proved the protective activity of rPly against S. pneumoniae serotype 3 at intranasal infection.

Aluminum hydroxide has long been employed as a vaccine adjuvant for its safety profile, although its precise mechanism of action remains elusive. In this study, we investigated the transcriptomic responses in sheep spleen following repetitive vaccination with aluminum adjuvanted vaccines and aluminum hydroxide alone. Notably, this work represents the first exploration of the sheep spleen transcriptome in such conditions. Animals were splitted in 3 treatment groups: vaccine group, adjuvant alone group and control group. A total of 18 high-depth RNA-seq libraries were sequenced, resulting in a rich dataset which also allowed isoform-level analysis. The comparisons between vaccine-treated and control groups (V vs C) as well as between vaccine-treated and adjuvant-alone groups (V vs A) revealed significant alterations in gene expression profiles, including protein coding genes and long non-coding RNAs. Among the differentially expressed genes, many were associated with processes such as endoplasmic reticulum (ER) stress, immune response and cell cycle. The analysis of co-expression modules further indicated a correlation between vaccine treatment and genes related to ER stress and unfolded protein response. Surprisingly, adjuvant-alone treatment had little impact on the spleen transcriptome. Additionally, the role of alternative splicing in the immune response was explored. We identified isoform switches in genes associated with immune regulation and inflammation, potentially influencing protein function. In conclusion, this study provides valuable insights into the transcriptomic changes in sheep spleen following vaccination with aluminum adjuvanted vaccines and aluminum hydroxide alone. These findings shed light on the molecular mechanisms underlying vaccine-induced immune responses and emphasize the significance of antigenic components in aluminum adjuvant mechanism of action. Furthermore, the analysis of alternative splicing revealed an additional layer of complexity in the immune response to vaccination in a livestock species.

In this study, a static mixer was used as an alternative to the existing flash mixing method for ballasted flocculation to assess the turbidity removal and ballasted floc formation characteristics. Synthetic magnetite exhibits excellent properties, such as high specific gravity, hydrophobicity, and wear resistance, making it a suitable ballast agent (BA). The experimental design was optimized using the response surface methodology. To evaluate turbidity removal, a model based on polyaluminum chloride dosage, BA surface charge, and pH was developed. To assess the ballasted floc characteristics, the BA dosage, BA size, and G value of the static mixer were used. During ballasted flocculation, the impact of the zeta potential of the BA was minimal. Consequently, bonding primarily resulted from the viscosity of the floc caused by physical collisions rather than electrostatic forces stemming from the BA charge. The findings of this study demonstrated promising outcomes, including potential energy savings and process streamlining, by identifying crucial design elements for implementing a static mixer in the ballasted flocculation process.

Protein-based subunit vaccines like RBD-Fc are promising tools to fight COVID-19. RBD-Fc fuses the receptor-binding domain (RBD) of the SARS-CoV-2 virus spike protein with the Fc region of human IgG1, making it more immunogenic than RBD alone. Earlier work showed that combining RBD-Fc with iNKT cell agonists as adjuvants improved neutralizing antibodies but did not sufficiently enhance T cell responses, a limitation RBD-Fc vaccines share with common adjuvants. Here we demonstrate that aluminum hydroxide combined with α-C-GC, a C-glycoside iNKT cell agonist, significantly improved the RBD-Fc vaccine\'s induction of RBD-specific T-cell responses. Additionally, aluminum hydroxide with α-GC-CPOEt, a phosphonate diester derivative, synergistically elicited more robust neutralizing antibodies. Remarkably, modifying αGC with phosphate (OPO3H2) or phosphonate (CPO3H2) to potentially enhance aluminum hydroxide interaction did not improve efficacy over unmodified αGC with aluminum hydroxide. These findings underscore the straightforward yet potent potential of this approach in advancing COVID-19 vaccine development and provide insights for iNKT cell-based immunotherapy.

To recycle aluminum (Al) from waterworks sludge resulting from polyaluminum chloride (PAC) used as coagulants, this study proposed an innovative strong acidic cation (SAC) exchange resin treatment strategy for Al separation from coexisting fulvic acid (FA) and heavy metals (HMs) in the H2SO4 leachate of PAC sludge. Fluorescence titration confirmed the breakdown of the Al-FA complex at pH 2.0, which facilitated Al separation from FA in the acidic leachate. The species distribution of the dissociated Al (i.e. Ala, Alb, and Alc) significantly influenced the adsorption of Al onto the cation exchange resin. The continuous release of H+ during the cation exchange reaction greatly promoted the transformation of dissociated Alc and Alb into Ala, thereby improving the adsorption of total Al. Moreover, the SAC resin column successfully separated the codissolved HMs from the Al in the leachate even at an influent pH of 2.8, which was attributed to the greater selectivity of the sulfonate groups on the cation exchange resin for free Al3+. The Al eluted from the exhausted resin with 1.1 M H2SO4 was collected as the recycled coagulant after proper pH adjustment. The Al adsorption capacity of the SAC resin decreased by approximately 5 % with each operation cycle and was regained by complete regeneration with 1.8 M H2SO4 after 5 cycles. Overall, the integrated efficiency of Al recovery from PAC sludge by H2SO4 acidification and SAC resin separation/purification reached 70.10 %. The recycled Al from sludge has a water treatment performance comparable to that of fresh PAC coagulant.

Clay-algae flocculation is a promising method to remove harmful algal blooms (HABs) in aquatic ecosystems. Many HAB-generating species, such as Microcystis aeruginosa (M. aeruginosa), a common species in lakes, produce toxins and harm the environment, human health, and the economy. Natural clays, such as bentonite and kaolinite, and modification of these clays have been applied to mitigate HABs by forming large aggregates and settling down. In this study, we aim to examine the impact of laponite, a commercially available smectite clay that is synthetic, transparent, compatible with human tissues, and degradable, on removing HABs. We compare the cell removal efficiencies (RE) of laponite, two natural clays, and their polyaluminum chloride (PAC)-modified versions through clay-algae flocculation experiments. Our results show that the optimum concentrations of laponite, bentonite, kaolinite, PAC-modified bentonite, and PAC-modified kaolinite to remove 80 % of the M. aeruginosa cells from the water column are 0.05 g/L, 2 g/L, 4 g/L, 2 g/L and 0.3 g/L respectively. Therefore, to achieve the same cell removal efficiency, the amount of laponite needed is 40 to 80 times less than bentonite and kaolinite, and 6 times less than PAC-modified kaolinite. We demonstrate that the superior performance of laponite clay is because of its smaller particle size, which increases the encounter rate between cells and clay particles. Furthermore, experiments using water samples from Powderhorn Lake confirmed laponite\'s effectiveness in mitigating HABs. Our price analysis also suggests that this commercially-available clay, laponite, can be used in the field at a relatively low cost.

Conjugation to carrier proteins is necessary for peptides to be able to induce antibody formation when injected into animals together with a suitable adjuvant. This is usually performed by conjugation in solution followed by mixing with the adjuvant. Alternatively, the carrier may be adsorbed onto a solid support followed by activation and conjugation with the peptide by solid-phase chemistry. Different reagents can be used for conjugation through peptide functional groups (-SH, -NH2, -COOH), and various carrier proteins may be used depending on the peptides and the intended use of the antibodies. The solid phase may be an ion exchange matrix, from which the conjugate can subsequently be eluted and mixed with adjuvant. Alternatively, the adjuvant aluminum hydroxide may be used as the solid-phase matrix, whereupon the carrier is immobilized and conjugated with peptide. The resulting adjuvant-carrier-peptide complexes may then be used directly for immunization.

Membrane fouling is a bottleneck issue that hindered the further application of ultrafiltration technology. To alleviate membrane fouling, coagulation-ultrafiltration (C-UF) process using polyaluminum chloride (PACl) and PACl-Al13 with high proportion of Al13O4(OH)247+ as coagulants, respectively, were investigated at various pH conditions. Results indicated that an increase in solution pH contributed to larger floc size and looser floc structure for both PACl and PACl-Al13. It was conducive to the formation of more porous cake, as evidenced by mean pore area and pore area distribution of cake, leading to lower reversible fouling. Furthermore, humic acid (HA) removal presented a trend of first increasing and then decreasing with the increase of pH. The optimal HA removal was achieved at pH 6 regardless of coagulant type, suggesting that the slightest irreversible fouling should be occurred at this point. Interestingly, the irreversible fouling with PACl coagulant achieved a minimum value at pH 9, while the minimal irreversible fouling with PACl-Al13 was observed at pH 6. We speculated that the cake formed by PACl could further intercept HA prior to UF process at alkaline pH. Furthermore, compared with PACl, PACl-Al13 had a stronger charge neutralization ability, thus contributing to more compact floc structure and higher HA removal at various pH conditions. By UF fractionation measurement, higher HA removal for PACl-Al13 was due to higher removal of HA with molecular weight less than 50 kDa.