Mixing Vessel Research Articles

Parallel single B cell transcriptomics to elucidate pig B cell repertoire

Pork is the most widely consumed meat on the planet, placing swine health as a critical factor for both the world economy and the food industry. Infectious diseases in pigs not only threaten these sectors but also raise zoonotic concerns, as pigs can act as “mixing vessels” for several animals and human viruses and can lead to the emergence of new viruses that are capable of infecting humans. Several efforts are ongoing to develop pig vaccines, albeit with limited success. This has been largely attributed to the complex nature of pig infections and incomplete understanding of the pig immune responses. Additionally, pig has been suggested to be a good experimental model to study viral infections (e.g., human influenza). Despite the significant importance of studying pig immunology for developing infection models, zoonosis, and the crucial need to develop better swine vaccines, there is still very limited information on the response of the swine adaptive immune system to several emerging pathogens. Particularly, very little is known about the pig B cell repertoire upon infection. Understanding the B cell repertoire is especially crucial towards designing better vaccines, predicting zoonosis and can provide insights into developing new diagnostic agents. Here, we developed methods for performing parallel single pig B cell (up to 10,000 B cells) global and immunoglobulin transcriptome sequencing. We then adapted a computational pipeline previously built for human/mouse sequences, to now analyze pig sequences. This allowed us to comprehensively map the B cell repertoire and get paired antibody sequences from pigs in a single parallel sequencing experiment. We believe that these approaches will have significant implications for swine diseases, particularly in the context of swine mediated zoonosis and swine and human vaccine development.

Large-Scale Serological Survey of Influenza A Virus in South Korean Wild Boar (Sus scrofa).

In this comprehensive large-scale study, conducted from 2015 to 2019, 7,209 wild boars across South Korea were sampled to assess their exposure to influenza A viruses (IAVs). Of these, 250 (3.5%) were found to be IAV-positive by ELISA, and 150 (2.1%) by the hemagglutination inhibition test. Detected subtypes included 23 cases of pandemic 2009 H1N1, six of human seasonal H3N2, three of classical swine H1N1, 13 of triple-reassortant swine H1N2, seven of triple-reassortant swine H3N2, and seven of swine-origin H3N2 variant. Notably, none of the serum samples tested positive for avian IAV subtypes H3N8, H5N3, H7N7, and H9N2 or canine IAV subtype H3N2. This serologic analysis confirmed the exposure of Korean wild boars to various subtypes of swine and human influenza viruses, with some serum samples cross-reacting between swine and human strains, indicating potential infections with multiple IAVs. The results highlight the potential of wild boar as a novel mixing vessel, facilitating the adaptation of IAVs and their spillover to other hosts, including humans. In light of these findings, we recommend regular and frequent surveillance of circulating influenza viruses in the wild boar population as a proactive measure to prevent potential human influenza pandemics and wild boar influenza epizootics.

UV–VIS imaging-based investigation of API concentration fluctuation caused by the sticking behaviour of pharmaceutical powder blends

Surface powder sticking in pharmaceutical mixing vessels poses a risk to the uniformity and quality of drug formulations. This study explores methods for evaluating the amount of pharmaceutical powder mixtures adhering to the metallic surfaces. Binary powder blends consisting of amlodipine and microcrystalline cellulose (MCC) were used to investigate the effect of the mixing order on the adherence to the vessel wall. Elevated API concentrations were measured on the wall and within the dislodged material from the surface, regardless of the mixing order of the components. UV imaging was used to determine the particle size and the distribution of the API on the metallic surface. The results were compared to chemical maps obtained by Raman chemical imaging. The combination of UV and VIS imaging enabled the rapid acquisition of chemical maps, covering a substantially large area representative of the analysed sample. UV imaging was also applied in tablet inspection to detect tablets that fail to meet the content uniformity criteria. The results present powder adherence as a possible source of poor content uniformity, highlighting the need for 100% inspection of pharmaceutical products to ensure product quality and safety.

The avian influenza A virus receptor SA-α2,3-Gal is expressed in the porcine nasal mucosa sustaining the pig as a mixing vessel for new influenza viruses

Influenza A viruses (IAVs) originate from wild birds but have on several occasions jumped host barriers and are now also circulating in humans and mammals. The IAV host receptors (glycans with galactose linked to a sialic acid (SA) in an α2,3 or α2,6 linkage) are crucial host factors restricting inter-species transmission. In general, avian-origin IAVs show a preference for SA-α2,3 (avian receptor), whereas IAVs isolated from humans and pigs prefer SA-α2,6 (human receptor). N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) are the two major SAs. Neu5Ac is expressed in all species, whereas Neu5Gc is only expressed in a limited number of domestic species such as pigs and horses, but not in humans. Despite that previous studies have shown that the IAV host receptor distribution appears to be similar in pigs and humans, none of these studies have investigated the expression of Neu5Gc-α2,6 in situ in porcine tissues. Thus, the aim of this study was to elucidate the distribution of IAV host receptors expressed in the porcine respiratory tract and relate the expression to the viral tropism of diverse host-adapted IAVs.The IAV receptor (SA-α2,3 and SA-α2,6) distribution and the presence of specifically Neu5Gc-α2,6 in the porcine nasal, tracheal, and lung tissues was investigated by lectin histochemistry. Furthermore, IAV immunohistochemistry was performed on tissues from pigs experimentally infected with IAVs, either adapted to pigs or humans, to investigate the significance of the IAV host receptors and the tropism of the diverse host-adapted IAVs. We document for the first time the expression of the avian receptor on the surface of the porcine nasal mucosa and an equal expression of Neu5Ac- α2,6 and Neu5Gc-α2,6 on the surface of the tracheal epithelium and alveoli. In all IAV-infected pigs, we found a low amount of IAV-positive cells in the trachea despite a high expression of the human receptor. Cumulatively, these findings suggest that optimal IAV replication involves a complex interplay between the viruses and their host receptors and that there might be other less clearly defined host factors that determine the site of replication.

Influenza A virus antibodies in dogs, hunting dogs, and backyard pigs in Campeche, Mexico.

This study aimed to identify exposure to human, swine, and avian influenza A virus subtypes in rural companion and hunting dogs, backyard pigs, and feral pigs. The study took place in a region of southeastern Mexico where the sampled individuals were part of backyard production systems in which different domestic and wild species coexist and interact with humans. We collected blood samples from pigs and dogs at each of the sites. We used a nucleoprotein enzyme-linked immunosorbent assay to determine the exposure of individuals to influenza A virus. Haemagglutination inhibition was performed on the positive samples to determine the subtypes to which they were exposed. For data analysis, a binomial logistic regression model was generated to determine the predictor variables for the seropositivity of the individuals in the study. We identified 11 positive individuals: three backyard pigs, four companion dogs, and four hunting dogs. The pigs tested positive for H1N1 and H1N2. The dogs were positive for H1N1, H1N2, and H3N2. The model showed that dogs in contact with backyard chickens are more likely to be seropositive for influenza A viruses. We demonstrated the essential role hunting dogs could play as intermediate hosts and potential mixing vessel hosts when exposed to human and swine-origin viral subtypes. These results are relevant because these dogs interact with domestic hosts and humans in backyard systems, which are risk scenarios in the transmission of influenza A viruses. Therefore, it is of utmost importance to implement epidemiological surveillance of influenza A viruses in backyard animals, particularly in key animals in the transmission of these viruses, such as dogs and pigs.

Influenza risks arising from mixed intensive pig and poultry farms, with a spotlight on the United Kingdom

Diseases passed to humans from animals (zoonoses) constitute 75% of emerging infectious diseases. Farmed animals are considered a high zoonotic risk, especially poultry and pigs as evidenced by recent outbreaks of avian and swine influenza. This review sought to collate recent knowledge of the disease risks from keeping pigs and chickens intensively and in close proximity to each other. Recent knowledge on influenza viruses compounds the public health concerns; no longer are concerns about “mixing vessel” hosts limited to pigs, but several other animal species too at a high level of probability—most notably chickens and humans. More generally, scientific literature establishing positive associations between intensive animal farming, human population growth, reduced biodiversity, and increased zoonoses risks is abundant. This includes the publication of relevant systematic reviews. The collected scientific evidence on this issue is clear: there is exceptionally strong evidence for a link between low animal welfare levels and high zoonotic risks, exacerbated by animal crowding, low genetic diversity, compromised hygiene, and high animal stress levels which compromise immune systems. Based on this evidence, further industrialized animal farms—especially poultry and pig farms or a mix thereof, and particularly in areas that already have a high concentration of farmed animals—should not generally be permitted to proceed. Instead, efforts should concentrate on supporting arable agriculture (or transitions toward this) and de-intensifying remaining animal farms, in alignment with One Health/One Welfare approaches within which animal health and welfare are integral parts of any farming operation. Among numerous other factors, this would involve reducing stocking densities down to 11 kg/m2 (around five chickens/m2) for meat chickens, and down to one pig/1.5 m2 for pigs (assuming a 100 kg pig).

Evolution of influenza A viruses in exhibition swine and transmission to humans, 2013-2015.

Swine are a mixing vessel for the emergence of novel reassortant influenza A viruses (IAV). Interspecies transmission of swine-origin IAV poses a public health and pandemic risk. In the United States, the majority of zoonotic IAV transmission events have occurred in association with swine exposure at agricultural fairs. Accordingly, this human-animal interface necessitates mitigation strategies informed by understanding of interspecies transmission mechanisms in exhibition swine. Likewise, the diversity of IAV in swine can be a source for novel reassortant or mutated viruses that pose a risk to both swine and human health. In an effort to better understand those risks, here we investigated the epidemiology of IAV in exhibition swine and subsequent transmission to humans by performing phylogenetic analyses using full genome sequences from 272 IAV isolates collected from exhibition swine and 23 A(H3N2)v viruses from human hosts during 2013-2015. Sixty-seven fairs (24.2%) had at least one pig test positive for IAV with an overall estimated prevalence of 8.9% (95% CI: 8.3-9.6, Clopper-Pearson). Of the 19 genotypes found in swine, 5 were also identified in humans. There was a positive correlation between the number of human cases of a genotype and its prevalence in exhibition swine. Additionally, we demonstrated that A(H3N2)v viruses clustered tightly with exhibition swine viruses that were prevalent in the same year. These data indicate that multiple genotypes of swine-lineage IAV have infected humans, and highly prevalent IAV genotypes in exhibition swine during a given year are also the strains detected most frequently in human cases of variant IAV. Continued surveillance and rapid characterization of IAVs in exhibition swine can facilitate timely phenotypic evaluation and matching of candidate vaccine strains to those viruses present at the human-animal interface which are most likely to spillover into humans.

Experimental & simulative assessment of mixing quality for dry Li-Ion cathode production in an Eirich intensive mixer

We investigated the possibility to characterize the dry-mixing in Li-Ion battery production by a simple quality parameter, the bulk density. We present a dimensionless number that allows to calculate the bulk density of the mixture in dependence on mixing parameters. Suitability of the dry powders for cell production was investigated with electrical conductivity measurements. The highest powder conductivity of 1 S/m was measured for the highest mixing tool velocity of 25 m/s after a mixing time of 4000 mixing tool revolutions in the EL0.1 mixing vessel. This indicates that high energy mixing for a short mixing time can result in a beneficial electrical conductivity in the bulk. Longer mixing times generally led to lower powder conductivities for all investigated mixing tool speeds between 5 m/s and 25 m/s in both mixing vessels. The experimental results were incorporated into a DEM simulation model and analyzed further.

Design the mechanical–chemical reactor for oily wastewater treatment

This study explains the mechanical design of a mixer with a focus on the fluid forces generated by the fluid continuum in the mixing reactor on the impellers. The research demonstrates that the forces are caused by transient asymmetries in fluid flow that act on the mixing impeller. The mixer shaft and gear reducer receive these dynamic loads from the impeller blades. A general equation for the fluid force behavior can be developed. The research also stresses on the significance of the mechanical interaction of the mixing process with the mixing vessel and impeller. Four parameters were measured; impeller speed for coagulation and flocculation, mixing time for coagulation, and flocculation. The experimental results in the final section will explain that impellers with four blades provide higher oil removal than impellers with two blades in oily wastewater treatment.

Application of HB Impeller from Test Tube to Pilot Scale Mixing Vessel

Application of HB Impeller from Test Tube to Pilot Scale Mixing Vessel

Numerical Analysis of Viscous Polymer Resin Mixing Processes in High-Speed Blade-Free Planetary Blender Using Smoothed Particle Hydrodynamics

High-speed planetary mixers can rapidly and efficiently combine rheological liquids, such as polymer resins and paste materials, because of the large centrifugal forces generated by the planetary motion of the mixing vessel. Only a few attempts have been made to computationally model and analyze the intricate mixing patterns of highly viscous substances. This paper presents meshless flow simulations of the planetary mixing of polymeric fluids. This research utilized the smoothed particle hydrodynamics (SPH) approach for numerical calculations. This method has advantages over the finite-volume method, which is a grid-based computational technique, when it comes to modeling interfacial and free surface flow problems. Newtonian rheology and interfacial surface force models were used to calculate the dissipative forces in the partial differential momentum equation of fluid motion. Simulations of the flow of an uncured polyurethane resin were carried out while it was mixed in a planetary mixer, under various operating conditions. Simulations using SPH were able to accurately reproduce the intricate flow and blending pattern, providing insight into mixing mechanics and mixing index evolution characteristics according to operating conditions for the planetary mixing of polymeric fluids. The simulation results showed that the spiral band, which promotes the mixing performance, is densely and distinctively formed under high-speed operation conditions.

Use of positron emission particle tracking to assess mixing of a graphite-based lithium-ion anode slurry in an Eirich mixer

The mixing of electrode slurries is a key processing step during the manufacturing of lithium-ion batteries, and poor mixing can have a profound effect on final cell electrochemical performance. This is complicated by the evolving non-Newtonian rheology of these slurries. In this study, Positron Emission Particle Tracking (PEPT) is used to determine the fluid dynamics and dispersion of a model graphite-based anode slurry in an Eirich EL1 mixer, equipped with a rotating pan, wall scraper and internal rotor. The main processes studied are the wetting of graphite particles and breakup of large graphite agglomerates. For experiments performed at different internal rotor tip speeds, with fixed outer pan speed, the dispersion dynamics change with the lowest rotor speed (2 m/s) leaving a large channel of poor mixing between the rotor region and the rotating pan, whilst the mixing within the vessel is mainly driven by the rotating pan. At higher mixing speeds (6 and 10 m/s) more of the vessel volume is active and there is no longer a poor mixing region observed, with 10 m/s tip speed producing the highest intensity of dispersion. An increasing mixer effectiveness (average dispersion across the mixing vessel) as well as an increasing dispersion co-efficient (rate of mixing) is also observed, which will be use to inform scale-up criteria.

Characterisation of oscillations in multi-phase unbaffled mixing vessels

Unbaffled mixing systems have several benefits over conventional baffled systems, including significantly lower power required for solids suspension. However, one drawback of unbaffled systems is the possibility of oscillations occurring within the tank. Experimental laboratory-scale experiments have revealed oscillations in settling solids and large variations in the input power, despite the impeller speed remaining constant. These oscillations have been found to be linked through having the same frequency. The frequency and amplitude of these oscillations has been determined across a range of agitator and tank configurations, including variations in impeller size and type, impeller clearance and submerged depth, and the surface boundary conditions. The effect of solids concentration has also been investigated. The oscillations were found to be inherently multi-phase and had a strong concentration dependence. Impeller size and type influenced the amplitude and frequency of the oscillations, as did the impeller clearance. Liquid surface condition did not influence the oscillations. The oscillation frequency appears to be related to the circulation time in the tank. Although these oscillations fall into the definition of previously observed macro-instabilities, several distinguishing features are noted.

Effects of shear-thinning rheological properties on the energy loss characteristics of a biomass mixing vessel

This study investigated the irreversible energy losses in the different sections of propeller blades. To the best of our knowledge, this is the first study to consider the properties of a shear-thinning fluid in evaluating irreversible energy losses based on the entropy generation theory. The numerical simulation results were consistent with the experimental results. The flow energy losses and the total mechanical energy loss gradient of an anaerobic digestion (AD) system were determined. The results indicated that the total mechanical energy loss occurred in the propeller region and was primarily influenced by the operation speed. The effects of rheology were neglected, although rheology notably affects the equivalent-volume velocity field within specific power characteristics, leading to an insufficient mixing field in the AD system. The energy losses primarily occurred around the propeller region, primarily in sections 3–5 under different flow rates. Viscous diffusion and velocity fluctuation are the primary factors contributing to the entropy of the system, accounting for more than 98%. According to the wall separation and friction loss on the suction and pressure surfaces of the propellers, sections 3–5 accounted for 90% of the energy loss. Energy dissipation in the propeller was mostly constituted by turbulence entropy and direct entropy. The rotation speed was the key factor causing viscous diffusion. Although the rheology effect on hydraulic loss is limited at low concentrations, the hydraulic loss in the blade tip region due to high-concentration fluids is significantly affected by rheology.

Heterogeneity and Recombination of Foot-and-Mouth Disease Virus during Multi-Strain Coinfection of Cattle.

Superinfection of cattle persistently infected with foot-and-mouth disease virus (FMDV), with a heterologous FMDV strain has been shown to generate novel recombinant viruses. In this study, we investigated the pathogenesis events within specific tissues associated with FMDV coinfections in cattle subjected to either simultaneous or serial exposure to two distinct strains of FMDV. Both strains of FMDV (one each of serotypes O and A) were similarly localized to the nasopharyngeal mucosa during the early stages of infection. However, while no recombinant FMDV genomes were recovered from simultaneously coinfected cattle, interserotypic recombinants were isolated from nasopharyngeal tissue samples obtained at 48 h after heterologous superinfection of a persistently infected FMDV carrier. Additionally, analysis of FMDV genomes obtained from replicate nasopharyngeal tissue samples demonstrated that adjacent segments of the mucosa were sometimes infected by distinct viruses, demonstrating a multifocal and heterogeneous distribution of FMDV infection during primary and persistent phases of infection. This work indicates that superinfection of FMDV carriers may be an important source of emergent recombinant strains of FMDV in areas where multiple strains are co-circulating. IMPORTANCE Foot-and-mouth disease (FMD) is a socioeconomically impactful livestock disease with a complex epidemiology and ecology. Although recombinant viruses have been identified in field samples, the mechanisms of emergence of those viruses have never been elucidated. This current study demonstrates how serial infection of cattle with two distinct serotypes of FMD virus (FMDV) leads to rapid generation of recombinant viruses in the upper respiratory tracts of infected animals. This finding is particularly relevant in relation to the management of persistently infected FMDV carrier cattle that can maintain subclinical FMDV infection for months to years after an initial infection. Such carrier animals may function as mixing vessels that facilitate the emergence of novel recombinant FMDV strains in areas where multiple virus strains are in circulation.

Zoonotic Animal Influenza Virus and Potential Mixing Vessel Hosts.

Influenza viruses belong to the family Orthomyxoviridae with a negative-sense, single-stranded segmented RNA genome. They infect a wide range of animals, including humans. From 1918 to 2009, there were four influenza pandemics, which caused millions of casualties. Frequent spillover of animal influenza viruses to humans with or without intermediate hosts poses a serious zoonotic and pandemic threat. The current SARS-CoV-2 pandemic overshadowed the high risk raised by animal influenza viruses, but highlighted the role of wildlife as a reservoir for pandemic viruses. In this review, we summarize the occurrence of animal influenza virus in humans and describe potential mixing vessel or intermediate hosts for zoonotic influenza viruses. While several animal influenza viruses possess a high zoonotic risk (e.g., avian and swine influenza viruses), others are of low to negligible zoonotic potential (e.g., equine, canine, bat and bovine influenza viruses). Transmission can occur directly from animals, particularly poultry and swine, to humans or through reassortant viruses in "mixing vessel" hosts. To date, there are less than 3000 confirmed human infections with avian-origin viruses and less than 7000 subclinical infections documented. Likewise, only a few hundreds of confirmed human cases caused by swine influenza viruses have been reported. Pigs are the historic mixing vessel host for the generation of zoonotic influenza viruses due to the expression of both avian-type and human-type receptors. Nevertheless, there are a number of hosts which carry both types of receptors and can act as a potential mixing vessel host. High vigilance is warranted to prevent the next pandemic caused by animal influenza viruses.

Can Acanthamoeba Harbor Monkeypox Virus?

Acanthamoeba is well known to host a variety of microorganisms such as viruses, bacteria, protozoa, and yeast. Given the recent number of cases of monkeypox infection, we speculate that amoebae may be aiding viral transmission to the susceptible hosts. Although there is no confirmatory evidence to suggest that Acanthamoeba is a host to monkeypox (a double-stranded DNA virus), the recent discovery of mimivirus (another double-stranded DNA virus) from Acanthamoeba, suggests that amoebae may shelter monkeypox virus. Furthermore, given the possible spread of monkeypox virus from animals to humans during an earlier outbreak, which came about after patients came in contact with prairie dogs, it is likely that animals may also act as mixing vessel between ubiquitously distributed Acanthamoeba and monkeypox virus, in addition to the environmental habitat that acts as an interface in complex interactions between diverse microorganisms and the host.

A recent outbreak of human H1N2 infection: correspondence.

A recent outbreak of human H1N2 infection: correspondence.

Isolation and characterization of mammalian orthoreovirus from bats in the United States.

Mammalian orthoreovirus (MRV) infects many mammalian species including humans, bats, and domestic animals. To determine the prevalence of MRV in bats in the United States, we screened more than 900 bats of different species collected during 2015 to 2019 by a real-time RT-PCR assay; 4.4% bats tested MRV-positive and 13 MRVs were isolated. Sequence and phylogenetic analysis revealed that these isolates belonged to four different strains/genotypes of viruses in serotypes 1 or 2, which contain genes similar to those of MRVs detected in humans, bats, bovine, and deer. Further characterization showed that these four MRV strains replicated efficiently on human, canine, monkey, ferret and swine cell lines. The 40/Bat/USA/2018 strain belonging to the serotype 1 demonstrated the ability to infect and transmit in pigs without prior adaptation. Taken together, this is evidence for different genotypes and serotypes of MRVs circulating in U.S. bats, which can be a mixing vessel of MRVs that may spread to other species, including humans, resulting in cross-species infections. This article is protected by copyright. All rights reserved.

A particle scale mixing measurement method using a generalized nearest neighbor mixing index

The dependence of mixing measurement on the sampling method and scale is a major concern in the characterization of the state of homogeneity of solid particle mixtures. A novel “particle neighborhood” based mixing index, called the generalized nearest neighbor (GNN) mixing index, is developed to measure solids mixing at the particle scale. GNN is a statistically robust, grid-independent index and provides reliable particle scale mixture homogeneity values. The GNN index has a further advantage that it can be readily adapted to mixtures of unequal proportions or mixtures containing more than two species. To test the GNN index, X-ray computed tomography (CT) images are obtained to noninvasively extract detailed particle distribution of binary mixtures consisting of different-density particles in the mixing vessel. CT images are acquired at different mixing stages to accurately describe mixture homogeneity evolution during the mixing process. Mixture homogeneity values are quantified using the novel GNN mixing index, and these values are compared with measurements obtained using different mixing indices, including a standard deviation-based mixing index and a grid-independent location-based mixing index. The GNN mixing index is found to be well-suited for reliable mixture homogeneity reporting at the particle scale.