Ceramic absorbed with calcium bicarbonate mesoscopic crystals partially inactivate scrapie prions.

Prion diseases are fatal neurodegenerative disorders affecting both humans and animals. The causative agent, prion, is extremely resistant to common disinfection procedures. Thus, effective prion inactivation strategies using relatively safe and less corrosive disinfectants are required. A solution containing CAC-717, mesoscopic crystals of calcium bicarbonate, exerts both antimicrobial and virucidal activities without apparent harmful effects. This study demonstrated that combined treatment with CAC-717 absorbed on ceramic (CAC-717 ceramic) and sodium dodecyl sulfate (SDS) substantially reduced the protein misfolding cyclic amplification (PMCA) seeding activity of Chandler strain scrapie mouse-brain homogenates (ScBH). Additionally, bioassays demonstrated that ScBH-inoculated mice treated with CAC-717 ceramic in combination with sodium dodecyl sulfate (SDS) did not develop disease. Furthermore, this combination effectively inactivated PMCA seeding activity on ScBH-coated stainless-steel wires below the detection limit. Overall, the findings suggest that combined treatment with CAC-717 ceramic and SDS represents a promising and less damaging approach for prion inactivation.

Enhanced sulfide removal by gas stripping in a novel reactor for anaerobic wastewater treatment.

Removal of sulfide by gas stripping using biogas produced in an internal phase-separated reactor (IPSR) was evaluated during anaerobic treatment. The IPSR consisted of upper and lower segments with a gas-liquid partitioning (GLP) valve between the sections. Wastewater was fed to the upper segment in the first stage and then to the lower segment in the second stage. The GLP valve separated the liquid phase from the gaseous phase and supplied biogas from the lower segment to the upper segment. The IPSR and a control reactor were fed with synthetic wastewater and operated in parallel under an organic loading rate of 12 kg COD/(m3 day) at 35 °C. The sulfide concentration increased to 400-600 mg S/L, which is above the previously reported 50% inhibition level for methanogenic activity. The IPSR showed higher H2S removal performance than the control reactor and removed approximately twice the H2S as the control reactor at 400 mg S/L, indicating that it can be used for the stable treatment of wastewater containing high concentrations of sulfide.

Recent Advances in Prion Inactivation by Plasma Sterilizer.

Prions, which cause transmissible spongiform encephalopathies (TSEs), are a notorious group of infectious agents with possibly the highest resistance to complete inactivation. Although various gas plasma instruments have been developed, studies on prion inactivation using gas plasma instruments are limited. Among them, the hydrogen peroxide gas plasma instrument, STERRAD® (Advanced Sterilization Products; ASP, Johnson & Johnson, Irvine, CA, USA), is recommended for prion inactivation of heat-sensitive medical devices. However, STERRAD® is not a plasma sterilizer but a hydrogen peroxide gas sterilizer. In STERRAD®, plasma generated by radio frequency (RF) discharge removes excess hydrogen peroxide gas and does not contribute to sterilization. This is also supported by evidence that the instrument was not affected by the presence or absence of RF gas plasma. However, recent studies have shown that other gas plasma instruments derived from air, nitrogen, oxygen, Ar, and a mixture of gases using corona, dielectric barrier, microwave, and pulse discharges can inactivate scrapie prions. As inactivation studies on prions other than scrapie are limited, further accumulation of evidence on the effectiveness of gas plasma using human-derived prion samples is warranted for practical purposes.

Differences in the isotopic signature of activated sludge in four types of advanced treatment processes at a municipal wastewater treatment plant.

The natural abundance of stable isotopes is a powerful tool for evaluating biological reactions and process conditions. However, there are few stable isotope studies on the wastewater treatment process. This study carried out the first investigation on variations in natural abundance of carbon and nitrogen stable isotope ratios (δ13C and δ15N) of activated sludge in four types of advanced treatment process (extended aeration activated sludge (EAAS), aerobic-anoxic-aerobic (A2O), recycled nitrification-denitrification (RND), and modified Bardenpho (MB)) at a municipal wastewater treatment plant. The δ13C and δ15N values of influent suspended solids settled in the primary sedimentation tank (i.e., primary sludge) ranged from -25.4‰ to -24.6‰ and 0.5‰-2.9‰, respectively, during monitoring periods. The δ13C values of the activated sludge were -24.6‰ to -23.6‰ (EAAS), -25.4‰ to -24.3‰ (A2O), -25.7‰ to -24.9‰ (RND), and -25.7‰ to -24.3‰ (MB). The δ13C values of the activated sludge were similar to those of influent suspended solids. However, the δ13C values of activated sludge in EAAS was significantly higher than in A2O, RND, and MB. Meanwhile, the δ15N values of activated sludge were obviously higher than influent suspended solids; 5.8‰-7.5‰ (EAAS), 6.6‰-8.1‰ (A2O), 5.5‰-7.5‰ (RND), and 5.3‰-7.6‰ (MB). Changes in δ13C and δ15N values of the activated sludge within the treatment system were also found. These findings indicate that changes in δ13C and δ15N values of the activated sludge rely on important function for biological wastewater treatment such as nitrification, denitrification, and methane oxidation through wastewater treatment over time.

Introduction to Current Progress in Advanced Research on Prions.

Prion diseases or transmissible spongiform encephalopathies (TSEs) are fatal neurological diseases that include Creutzfeldt-Jakob disease (CJD) in humans, scrapie in sheep and goats, bovine spongiform encephalopathy (BSE) in cattle, camel spongiform encephalopathy (CSE) in camels and chronic wasting disease (CWD) in cervids. A key event in prion diseases is the conversion of the cellular, host-encoded prion protein (PrPC) to its abnormal isoform (PrPSc) predominantly in the central nervous system of the infected host (Aguzzi et al., 2004). These diseases are transmissible under some circumstances, but unlike other transmissible disorders, prion diseases can also be caused by mutations in the host gene. The mechanism of prion spread among sheep and goats that develop natural scrapie is unknown. CWD, transmissible mink encephalopathy (TME), BSE, feline spongiform encephalopathy (FSE), and exotic ungulate encephalopathy (EUE) are all thought to occur after the consumption of prion-infected material. Most cases of human prion disease occur from unknown reasons, and greater than 20 mutations in the prion protein (PrP) gene may lead to inherited prion disease. In other instances, prion diseases are contracted by exposure to prion infectivity. These considerations raise the question of how a mere protein aggregate can bypass mucosal barriers, circumvent innate and adoptive immunity, and traverse the blood-brain barrier to give rise to brain disease. Here, we will briefly introduce a few topics in current prion studies.

Function of Prion Protein and the Family Member, Shadoo.

Lowering cellular prion protein (PrPC) levels in the brain is predicted to be a powerful therapeutic strategy for the prion disease. PrPC may act as an antiapoptotic agent by blocking some of the internal environmental factors that initiate apoptosis. Prion protein (PrP)-knockout methods provide powerful indications on the neuroprotective function of PrPC. Using PrPC-knockout cell lines, the inhibition of apoptosis through stress inducible protein1 (STI1) is mediated by PrPC-dependent superoxide dismutase (SOD) activation. Besides, PrP-knockout exhibited wide spread alterations of oscillatory activity in the olfactory bulb as well as altered paired-pulse plasticity at the dendrodendric synapse. Both the behavioural and electro-physiological phenotypes could be rescued by neuronal PrPC expression. Neuprotein Shadoo (Sho), similarly to PrPC, can prevent neuronal cell death induced by the expression of PrP△HD mutants, an artificial PrP mutant devoid of internal hydrophobic domain. Sho can efficiently protect cells against exito-toxin-induced cell death by glutamates. Sho and PrP seem to be dependent on similar domains, in particular N-terminal (N), and their internal hydrophobic domain. Sho△N and Sho△HD displayed a reduced stress-protective activity but are complex glycosylated and attached to the outer leaflet of the plasma membrane via glycosylphosphatidylinositol (GPI) anchor indicating that impaired activity is not due to incorrect cellular trafficking. In Sho, over-expressed mice showed large amyloid plaques not seen in wild-type mice. However, Shadoo is not a major modulator of abnormal prion protein (PrPSc) accumulation. Sho and PrP share a stress-protective activity. The ability to adopt a toxic conformation of PrPSc seems to be specific for PrP.

Effect of Microglial Inflammation in Prion Disease.

Prion diseases are a group of transmissible fatal neurodegenerative disorders. Neuropatho- logical features of prion diseases include neuroinflammation featuring the infiltration of activated microglia in affected brain areas as well as the accumulation of an abnormal isoform of the cellular prion protein and neuronal loss. Recent studies have elucidated that inflammation in the brain induced by microglia plays an important role in the pathogenesis of neurodegenerative disorders including prion disease. Thus, the regulation of neuroin- flammation is key in terms of therapeutic and preventative approaches. The functions of neuroinflammation and microglia in this disease are discussed in this article.

The diabetes pandemic and associated infections: suggestions for clinical microbiology.

There are 425 million people with diabetes mellitus in the world. By 2045, this figure will grow to over 600 million. Diabetes mellitus is classified among noncommunicable diseases. Evidence points to a key role of microbes in diabetes mellitus, both as infectious agents associated with the diabetic status and as possible causative factors of diabetes mellitus. This review takes into account the different forms of diabetes mellitus, the genetic determinants that predispose to type 1 and type 2 diabetes mellitus (especially those with possible immunologic impact), the immune dysfunctions that have been documented in diabetes mellitus. Common infections occurring more frequently in diabetic vs. nondiabetic individuals are reviewed. Infectious agents that are suspected of playing an etiologic/triggering role in diabetes mellitus are presented, with emphasis on enteroviruses, the hygiene hypothesis, and the environment. Among biological agents possibly linked to diabetes mellitus, the gut microbiome, hepatitis C virus, and prion-like protein aggregates are discussed. Finally, preventive vaccines recommended in the management of diabetic patients are considered, including the bacillus calmette-Guerin vaccine that is being tested for type 1 diabetes mellitus. Evidence supports the notion that attenuation of immune defenses (both congenital and secondary to metabolic disturbances as well as to microangiopathy and neuropathy) makes diabetic people more prone to certain infections. Attentive microbiologic monitoring of diabetic patients is thus recommendable. As genetic predisposition cannot be changed, research needs to identify the biological agents that may have an etiologic role in diabetes mellitus, and to envisage curative and preventive ways to limit the diabetes pandemic.

Disinfection and Sterilization Using Plasma Technology: Fundamentals and Future Perspectives for Biological Applications.

Recent studies have shown that plasma can efficiently inactivate microbial pathogens such as bacteria, fungi, and viruses in addition to degrading toxins. Moreover, this technology is effective at inactivating pathogens on the surface of medical and dental devices, as well as agricultural products. The current practical applications of plasma technology range from sterilizing therapeutic medical devices to improving crop yields, as well as the area of food preservation. This review introduces recent advances and future perspectives in plasma technology, especially in applications related to disinfection and sterilization. We also introduce the latest studies, mainly focusing on the potential applications of plasma technology for the inactivation of microorganisms and the degradation of toxins.

PrP Knockout Cells Expressing Transmembrane PrP Resist Prion Infection.

Glycosylphosphatidylinositol (GPI) anchoring of the prion protein (PrPC) influences PrPC misfolding into the disease-associated isoform, PrPres, as well as prion propagation and infectivity. GPI proteins are found in cholesterol- and sphingolipid-rich membrane regions called rafts. Exchanging the GPI anchor for a nonraft transmembrane sequence redirects PrPC away from rafts. Previous studies showed that nonraft transmembrane PrPC variants resist conversion to PrPres when transfected into scrapie-infected N2a neuroblastoma cells, likely due to segregation of transmembrane PrPC and GPI-anchored PrPres in distinct membrane environments. Thus, it remained unclear whether transmembrane PrPC might convert to PrPres if seeded by an exogenous source of PrPres not associated with host cell rafts and without the potential influence of endogenous expression of GPI-anchored PrPC To further explore these questions, constructs containing either a C-terminal wild-type GPI anchor signal sequence or a nonraft transmembrane sequence containing a flexible linker were expressed in a cell line derived from PrP knockout hippocampal neurons, NpL2. NpL2 cells have physiological similarities to primary neurons, representing a novel and advantageous model for studying transmissible spongiform encephalopathy (TSE) infection. Cells were infected with inocula from multiple prion strains and in different biochemical states (i.e., membrane bound as in brain microsomes from wild-type mice or purified GPI-anchorless amyloid fibrils). Only GPI-anchored PrPC supported persistent PrPres propagation. Our data provide strong evidence that in cell culture GPI anchor-directed membrane association of PrPC is required for persistent PrPres propagation, implicating raft microdomains as a location for conversion. IMPORTANCE: Mechanisms of prion propagation, and what makes them transmissible, are poorly understood. Glycosylphosphatidylinositol (GPI) membrane anchoring of the prion protein (PrPC) directs it to specific regions of cell membranes called rafts. In order to test the importance of the raft environment on prion propagation, we developed a novel model for prion infection where cells expressing either GPI-anchored PrPC or transmembrane-anchored PrPC, which partitions it to a different location, were treated with infectious, misfolded forms of the prion protein, PrPres We show that only GPI-anchored PrPC was able to convert to PrPres and able to serially propagate. The results strongly suggest that GPI anchoring and the localization of PrPC to rafts are crucial to the ability of PrPC to propagate as a prion.

Dual role of cellular prion protein in normal host and Alzheimer's disease.

Using PrPC-knockout cell lines, it has been shown that the inhibition of apoptosis through STI1 is mediated by PrPC-dependent SOD activation. Antioxidant PrPC may contribute to suppression of inflammasome activation. PrPC is functionally involved in copper metabolism, signal transduction, neuroprotection, and cell maturation. Recently several reports have shown that PrPC participates in trans-membrane signaling processes associated with hematopoietic stem cell replication and neuronal differentiation. In another role, PrPC also tends to function as a neurotoxic protein. Aß oligomer, which is associated with neurodegeneration in Alzheimer's disease (AD), has also been reported to act as a ligand of PrPC. However, the physiological role of PrPC as an Aß42-binding protein is not clear. Actually, PrPC is critical in Aß42-mediated autophagy in neurons. PrPC shows a beneficial role in lipid rafts to promote autophagy. Further search for PrPC-interaction molecules using Prnp-/- mice and various types of Prnp-/- cell lines under various conditions may elucidate other important PrPC important functions.

Evaluation of the process performance of a down-flow hanging sponge reactor for direct treatment of domestic wastewater in Bangkok, Thailand.

This study assesses the performance of an aerobic trickling filter, down-flow hanging sponge (DHS) reactor, as a decentralized domestic wastewater treatment technology. Also, the characteristic eukaryotic community structure in DHS reactor was investigated. Long-term operation of a DHS reactor for direct treatment of domestic wastewater (COD = 150-170 mg/L and BOD = 60-90 mg/L) was performed under the average ambient temperature ranged from 28°C to 31°C in Bangkok, Thailand. Throughout the evaluation period of 550 days, the DHS reactor at a hydraulic retention time of 3 h showed better performance than the existing oxidation ditch process in the removal of organic carbon (COD removal rate = 80-83% and BOD removal rate = 91%), nitrogen compounds (total nitrogen removal rate = 45-51% and NH4+-N removal rate = 95-98%), and low excess sludge production (0.04 gTS/gCOD removed). The clone library based on the 18S ribosomal ribonucleic acid gene sequence revealed that phylogenetic diversity of 18S rRNA gene in the DHS reactor was higher than that of the present oxidation ditch process. Furthermore, the DHS reactor also demonstrated sufficient COD and NH4+-N removal efficiency under flow rate fluctuation conditions that simulates a small-scale treatment facility. The results show that a DHS reactor could be applied as a decentralized domestic wastewater treatment technology in tropical regions such as Bangkok, Thailand.

Evaluation of the resilience of a full-scale down-flow hanging sponge reactor to long-term outages at a sewage treatment plant in India.

Resilience to process outages is an essential requirement for sustainable wastewater treatment systems in developing countries. In this study, we evaluated the ability of a full-scale down-flow hanging sponge (DHS) reactor to recover after a 10-day outage. The DHS tested in this study uses polyurethane sponge as packing material. This full-scale DHS reactor has been tested over a period of about 4 years in India with a flow rate of 500 m(3)/day. Water was not supplied to the DHS reactor that was subjected to the 10-day outage; however, the biomass did not dry out because the sponge was able to retain enough water. Soon after the reactor was restarted, a small quantity of biomass, amounting to only 0.1% of the total retained biomass, was eluted. The DHS effluent achieved satisfactory removal of suspended solids, chemical oxygen demand, and ammonium nitrogen within 90, 45, and 90 min, respectively. Conversely, fecal coliforms in the DHS effluent did not reach satisfactory levels within 540 min; instead, the normal levels of fecal coliforms were achieved within 3 days. Overall, the tests demonstrated that the DHS reactor was sufficiently robust to withstand long-term outages and achieved steady state soon after restart. This reinforces the suitability of this technology for developing countries.

Protection of biomass from snail overgrazing in a trickling filter using sponge media as a biomass carrier: down-flow hanging sponge system.

This study investigated down-flow hanging sponge (DHS) technology as a promising trickling filter (TF) using sponge media as a biomass carrier with an emphasis on protection of the biomass against macrofauna overgrazing. A pilot-scale DHS reactor fed with low-strength municipal sewage was operated under ambient temperature conditions for 1 year at a sewage treatment plant in Bangkok, Thailand. The results showed that snails (macrofauna) were present on the surface of the sponge media, but could not enter into it, because the sponge media with smaller pores physically protected the biomass from the snails. As a result, the sponge media maintained a dense biomass, with an average value of 22.3 gVSS/L sponge (58.1 gTSS/L sponge) on day 370. The snails could graze biomass on the surface of the sponge media. The DHS reactor process performance was also successful. The DHS reactor requires neither chemical treatments nor specific operations such as flooding for snail control. Overall, the results of this study indicate that the DHS reactor is able to protect biomass from snail overgrazing.

Review of studies that have used knockout mice to assess normal function of prion protein under immunological or pathophysiological stress.

Deletion of cellular isoform of prion protein (PrP(C)) increases neuronal predisposition to damage by modulating apoptosis and the negative consequences of oxidative stress. In vivo studies have demonstrated that PrP(C)-deficient mice are more prone to seizure, depression, and induction of epilepsy and experience extensive cerebral damage following ischemic challenge or viral infection. In addition, adenovirus-mediated overexpression of PrP(C) reduces brain damage in rat models of cerebral ischemia. In experimental autoimmune encephalomyelitis, PrP(C)-deficient mice reportedly have a more aggressive disease onset and less clinical improvement during the chronic phase than wild-type mice mice. In mice given oral dextran sulfate, PrP(C) has a potential protective role against inflammatory bowel disease. PrP(C)-deficient mice demonstrate significantly greater increases in blood glucose concentrations after intraperitoneal injection of glucose than wild-type mice. Further in vivo challenges to PrP gene-deficient models and conditional knockout models with siRNA and in vivo administration of PrP-ligating agents may assist in refining knowledge of the lymphoid function of PrP(C) and predicting the effects of anti-PrP treatment on the immune system. Together, these findings indicate that PrP(C) may have multiple neuroprotective and anti-inflammatory roles, which explains why this protein is so widely expressed.

Pilot-scale experiment of down-flow hanging sponge for direct treatment of low-strength municipal wastewater in Bangkok, Thailand.

A pilot-scale experiment of a down-flow hanging sponge (DHS) reactor for treatment of low-strength municipal wastewater was conducted over 1 year in Bangkok, Thailand, to establish an appropriate method for treatment under tropical climate conditions. Municipal wastewater with an average BOD of 19 mg/L was fed directly into the DHS reactor. Superior effluent quality (5.1 ± 3.4 mg/L TSS, 21.1 ± 9.0 mg/L COD, 2.8 ± 1.4 mg/L BOD, and 4.1 ± 1.0 mg/L TN) was achieved at a hydraulic retention time (HRT) of 1 h under an average temperature of 30 °C. The DHS reactor reached an actual HRT of 19.0 min, indicating good contact efficiency between wastewater and retained sludge. The DHS reactor retained dense sludge at 15.3-26.4 g VSS/L based on the sponge media volume. The sludge activity in terms of specific oxygen uptake rate was good. Excess sludge was produced as 0.051 g TSS/g COD removed (0.11 g TSS/g BOD removed), and a good SVI of 28 mL/g was observed. The sufficient performance was attributed to dense sludge with high activity, regardless of the low-strength wastewater. Overall, the DHS was advantageous owing to its simple operation, lack of operational problems, and low power consumption.

Development of a down-flow hanging sponge reactor for the treatment of low strength sewage.

The process performance of a down-flow hanging sponge (DHS) reactor for treating low strength sewage (biochemical oxygen demand (BOD) 20-50 mg/L) was investigated in Bangkok, Thailand. The hydraulic retention time (HRT) was set at 4 h during the start-up period and was reduced to 1.5 h in a stepwise manner. Throughout the 300-day operational period, the DHS reactor shows high performance with respect to the removal of total suspended solid (>90% total suspended solid removal efficiency). No clogging of sponge media was observed in response to the self-digestion phenomena of the biofilm. At a HRT of 1.5 h, the BOD removal efficiency was sufficiently high (about 85%). The pathogen Escherichia coli and other coliform bacteria were removed almost completely as well (removal was 99.4% and 98.1%, respectively). Regarding the retained sludge activity measurement, the nitrite oxidation rate was higher than the ammonium oxidation rate (0.031 and 0.022 gram of nitrogen per gram of volatile suspended solids per day, respectively). In the 300 days of operation, the amount of excess sludge production was negligible. Thus, no sludge treatment system is required. Introduction of the DHS system in developing countries is recommended because this system requires a relatively small area, and has low electricity consumption and operation costs.

Development of appropriate technology for treatment of molasses-based wastewater.

In this study, the performance of a proposed treatment system consisting of an anaerobic process (acidification, methane fermentation) and an aerobic process (trickling filter) was evaluated for treating high concentrations of molasses-based wastewater (43-120 gCOD/L) by a continuous flow experiment. An anaerobic up-flow staged sludge bed (USSB) reactor, equipped with multiple gas solid separators, was used as the main treatment/methane recovery process. The USSB showed good efficiency of both COD removal (80-87%) and methane recovery (70-80%) at an organic loading rate of 11-43 kgCOD/m(3) day. As the influent COD concentration was increased, the organic loading rate for stable operation of the USSB was reduced due to cation inhibition. However, the COD removal efficiency of the whole treatment system (including the aerobic post-treatment process) was 96% even at an influent COD concentration of 120 gCOD/L. Use of the treated wastewater as a fertilizer and/or irrigation-water for sugarcane was evaluated by a field cultivation test. Both growth of sugarcane and emission of greenhouse gases from the field soil were measured. A relatively high methane flux (352 µgCH4/m(2) h) was observed when the treated wastewater from day 0 was used. By day 3, however, this value was reduced to the same level as the control. In addition, growth of sugarcane was satisfactory when the treated wastewater was used. The treated wastewater was found to be useful for cultivation of sugarcane in terms of both a low risk of greenhouse gas emission from the field soil and effectiveness for growth of sugarcane.

New Advances in Viral and Microorganism Disinfectants.

Nanomaterials are used to develop simpler, cheaper, and faster methods for disease diagnosis [...].

Antiviral agents and disinfectants for foot­and­mouth disease (Review).

Fluorouracil, 5-azacytidine, 6-azauridine, ribavirin, favipiravir (T-705) and its derivative (T-1105) exhibit anti-foot-and-mouth disease virus (FMDV) effects. In particular, T-1105 exhibits promising results when administered to guinea pigs orally, and pigs in their feed. FMDV is excreted in the early stages of infection in aerosols and oral or nasal droplets from animals. T-1105 along with the FMDV vaccine can be used to combat foot-and-mouth disease (FMD) epidemics. Several studies have shown that sodium hypochlorous solutions are widely used to inactivate viruses, including FMDV. However, these solutions must be stored under cool and dark conditions to maintain their virucidal effects. Interestingly, a study indicated that the virucidal activity of a calcium bicarbonate solution with a mesoscopic structure (CAC-717) did not decrease after storage at room temperature for at least four years outside direct sunlight. Numerous lessons acquired from the 2010 FMD outbreak in Japan are relevant for the control of COVID-19. However, the widespread use of chlorite can cause environmental issues. Chlorite can be combined with nitrogen to produce chloramine or N-nitrosodimethylamine, which plays a role in carcinogenesis. Therefore, risk assessments should be conducted in aquatic environments. Moreover, there is a need to develop nonchlorine disinfectants that can be used during epidemics, including FMD. The approach of 'One Health' should be shared between the public health and veterinary fields to improve the management of viral outbreaks, including those due to FMD.