Immersion In Bath Research Articles

Factors Affecting Post-Challenge Survival of Flavobacterium psychrophilum in Susceptible Rainbow Trout from the Literature.

Infectious bacterial pathogens are a concern for aquaculture as estimates suggest that billions of US dollars are lost annually in aquaculture due to disease. One of the most prevalent salmonid pathogens is the bacterium Flavobacterium psychrophilum that causes bacterial coldwater disease. We reviewed the published F. psychrophilum literature and conducted a Bayesian analysis to examine large-scale patterns in rainbow trout (Oncorhynchus mykiss) mortality associated with laboratory challenge. We incorporated factors that were common across a majority of the laboratory exposure studies and these included bacterial dose, culture time, exposure method, bacterial isolate, experimental duration, and fish weight. The comparison showed that injection as the exposure method produced higher mortality than bath immersion, bacterial isolates differed in their effect on mortality, and bacterial dose has an interactive effect with fish weight and exposure method. Our comparison allows for inference on factors affecting rainbow trout mortality due to exposure to F. psychrophilum and suggests avenues to further optimize research protocols to better reach study goals.

Raspberry pulp pasteurization: Computational fluid dynamics modeling and experimental validation of color and bioactive compound retention

AbstractThis work evaluated the heat transfer, fluid flow, and different reactions that occur during the pasteurization of raspberry pulp. The pulp was packed in glass jars and thermal processing was conducted by immersion in a water bath. The process was evaluated by multiphysics modeling, with the aim of determining the temperature distribution, flow pattern, the slowest heating zone (SHZ) location, the microbial inactivation, and the quality changes taking place during processing. In order to simulate the process accurately, a model was developed using experimentally measured product and heat transfer properties (density, thermal expansion, specific heat, thermal conductivity, apparent viscosity, and convective heat transfer coefficient) and kinetics of thermal degradation of quality parameters (color, anthocyanin, and ascorbic acid). The model was successfully validated against experimental measurements of both temperature and quality losses. After validation, the model was used to design and evaluate equivalent thermal processing conditions. Improved processes were identified that minimized the changes of color and the bioactive compounds, while maintaining product safety and stability.Practical ApplicationsThermal processing of raspberries may induce several physicochemical changes that impair their sensorial properties, and may reduce the content of nutrients and bioactive compounds. Hence, the design and optimization of heat treatments is a key tool to guarantee safety while, at the same time, minimizing detrimental side effects which involve a loss of quality. In this investigation, we assessed the actual thermo‐physical properties of raspberry pulp and the system heat transfer coefficients. Then, we developed a CFD model to predict the temperature and velocity transient profiles, the location of the SHZ, and the quality changes occurring in raspberry pulp during pasteurization. The findings of this study could be useful to food processors for optimizing pasteurization conditions in order to obtain safe and high‐quality raspberry pulp preserves, as well as for reducing processing energy requirements.

Graded concentrations of lidocaine hydrochloride in the modulation of behavioral, cardiac, and muscular responses of the Amazon freshwater fish tambaqui (Colossoma macropomum)

The present study was designed to characterize the behavior and evaluate the electromyographic and electrocardiographic responses of tambaqui (C. macropomum) exposed to different concentrations of lidocaine hydrochloride in immersion baths and after transfer to anesthetic-free water. For this purpose, fish juveniles (25.38 ± 6.5 g) were acclimated to aquariums for 15 days under controlled environmental and water quality conditions. Four concentrations of lidocaine hydrochloride (LA) (150, 175, 200, and 225 mg/L LA) were used in immersion baths to evaluate behavioral, electromyographic, and electrocardiographic changes during short-term anesthetic induction (5-min exposure) and in a washout. Upon exposure to LA, and irrespective of concentration, fish showed loss of the righting reflex, along with significant myorelaxation, which was partially achieved at concentrations of 150 and 175 mg/L LA and fully attained at 200 and 225 mg/L LA. Notably, a decreased heart rate and alteration of the T wave tracing occurred at higher concentrations. Notwithstanding, although protocols using LA at 225 mg/L or above could require greater care in monitoring, the prompt reversibility of such responses in recovering fish indicates that LA can be safely used in immersion baths for tambaqui C. macropomum as an alternative to other anesthetics, especially those in which important side effects have been documented.

Development of formalin killed vaccine candidate against streptococcosis caused by Enterococcus sp. in Nile tilapia

Multiple bacteria under Streptococcus, Enterococcus and Lactococcus genera are considered to be associated with Streptococcosis. This study was conducted to develop and evaluate the effectiveness of whole cell formalin killed vaccine prepared from E. faecalis, E. hirae and E. faecium against the streptococcosis in Nile tilapia. To develop the candidate vaccine, efficacy of whole cell killed E. faecalis (strains BFF1B1 & BFTS22), E. hirae (strain BFTS29) and E. faecium (strain BFTS31) were evaluated. The vaccine prepared by mixing 3 % formalin and left for 24 h to kill the bacterial cells. Physiological saline (0.85 %) was mixed to the harvested formalin free killed cells to make final suspension for vaccine. This vaccine was administered to the fingerling tilapia (average weight 30 ± 5 g) by interperitoneal injection (IP) and bath immersion (BI) methods. The vaccinated group was monitored for 28 days to assess the developed immunity and vaccine safety into the fish. Post vaccination in vivo challenge test were performed against the pathogens. The hematological parameters (RBC, WBC, platelets and hemoglobin) and specific IgM were found significantly higher (p < 0.001) in the vaccinated fish compared to those in the non-vaccinated fish. The mean mortality in IP and BI method ranged from 10.0 ± 0.00 % to 23.3 ± 5.77 % and 18.33 ± 5.30 % to 40.0 ± 0.10 %, respectively. The relative percentage survival (RPS) in IP and BI varied from 68.78 % to 88.57 % and 59.52 % to 69.05%, respectively. Mortality was significantly lower in the immunized fish compared to those in non-immunized fish. The RPS were not different (p > 0.05) between IP and BI vaccination method. Formalin killed vaccine showed an excellent efficacy against Enterococcus sp. infection in Nile tilapia by IP method although bath immersion could also be an alternative.

Analysis of Platinum Distribution within a Nafion 212 Membrane during Electroless Deposition

Ionic polymer metal composite (IPMC) is an electrically-responsive material made of perfluorinated sulfonic acid (PFSA) ionomers electroless plated with noble metal, such as platinum. Recently, interest in thin IPMC has been rising because of its potential applications in micro-electromechanical systems. However, little effort has been made so far on studying metal electroless deposition within thin PFSA membranes (thinner than 100 um). In this paper we study the platinum distribution within a thin Nafion membrane (Nafion 212, 50 µm-thick) using standard manufacturing recipe. We compare the obtained distribution with the distribution reported in literature based on thick Nafion membrane (Nafion 117, 180 µm-thick). We show that in Nafion 212 the maximum Pt concentration is situated in the bulk of the membrane, while in Nafion 117 the maximum peak is situated at the liquid-membrane boundary, suggesting a different rate determining deposition mechanism. Electroless deposition of platinum in Nafion 212 Ionic polymer metal composite (IPMC) is rising interest in the microelectromechanical system (MEMS) since smaller and thinner IPMC substrates would expand the applications. IPMCs are mostly made of a perfluorinated sulfonic acid ionomers (PFSA) membrane, such as commercially-available Nafion, coated with platinum. The prevalent way of manufacturing IPMC relies on the chemical reduction of platinum on the surface of the PFSA membrane, i.e. electroless deposition (1). Most of the reports on IPMC manufacturing rely on Nafion 117 (thickness of 0.180 mm, equivalent weight 1100 g/mol) while Nafion 212 has been poorly studied (thickness 0.050 mm, equivalent weight 1100 g/mol). This contribution aims at filling this gap.Materials and methodsA PFSA membrane (Nafion 212 membrane) has been coated with platinum using the standard recipe already describe elsewhere (1). An immersion-reduction step was performed in an immersion bath composed of tetraammineplatinum chloride (Pt(NH3)4Cl2, 0.2%), and reduction baths made of sodium borohydride (NaBH4, 5%). After the first immersion (30 min and 2 hours, respectively), the samples were immerged in the reduction bath until the end of the precipitation. Samples were then frozen in liquid nitrogen and cut, and analyzed with electrons dispersive x-ray spectroscopy (EDS) and processing of scanning electron microscopy (SEM) images (Fig 1a).Figure 1. a) EDS detection of platinum (yellow) overlapped with SEM picture of the cross-section of IPMC. Platinum distribution and Rayleigh fit for single reduction step (NaBH4) of platinum in a Nafion 212 membrane, after 30 minutes (b) and 2 hours (c) of immersion.Results and discussionThe distribution of platinum within Nafion 117 has already been studied by Millet et al. (2). The Pt distributions thereby reported strongly differs from the profiles we obtained (Fig 2). In the aforementioned profiles, the maximum Pt concentration is located at the liquid-substrate boundary of the Nafion membrane (2), whereas the Pt distribution in Nafion 212 reaches the maximum inside the Nafion membrane, between 0.5 µm and 1 µm below the surface (Fig 1b 1c).As reported by Millet et al., two limiting types of kinetics may be encountered in a dynamic exchange process, such that the deposition rate-determining mechanism is either diffusion in the liquid-film boundary (F-mechanism), or diffusion in the membrane (M-mechanism).The position of the peaks in our experimental data suggests that the mechanism determining the Pt precipitation rate inside Nafion 212 is diffusion inside the membrane while using Nafion 117 with the same reducing agent concentration leads to a precipitation rate determined by diffusion at the liquid/film boundary. It has been proven experimentally that ionic conductivity and water uptake of Nafion 212 is higher than for Nafion 117 (3). Thus we can hypothesize that the different behavior of thinner Nafion could explain the different empirical Pt distribution obtained using same reducing agent concentration. We could obtain a good fit of the Pt distribution (R2=0.919 and R2=0.945) within Nafion 212 with the Rayleigh distribution for immersion time of 30 minutes and 2 hours respectively (Eq 1.):f(x,σ 2) = (x/σ 2) exp(-x/2σ 2) [1]The scale parameter σ was found to be 4.19 and 4.58 for the sample immerged for 30 minutes and 2 hours, respectively. Further investigation of the precipitation mechanism inside Nafion 212 membrane and estimations of the phenomenological equations of mass transport correlated to Rayleigh distribution is reserved for future work.References K. Kim, M. Shahinpoor, Smart Mater. Struct., 12, 1, p. 65 (2003).P. Millet, R. Durand, E. Dartyge, G. Tourillon, and A. Fontaine J. Electrochem. Soc., 140, 5, pp. 1373-1380 (1993).A. Kusoglu, A.Z. Weber Chem. Rev. 117, 3 pp. 987-1104 (2017). Figure 1

Large scale immersion bath for isothermal testing of lithium-ion cells

Testing of lithium-ion batteries depends greatly on accurate temperature control in order to generate reliable experimental data. Reliable data is essential to parameterise and validate battery models, which are essential to speed up and reduce the cost of battery pack design for multiple applications. There are many methods to control the temperature of cells during testing, such as forced air convection, liquid cooling or conduction cooling using cooling plates. Depending on the size and number of cells, conduction cooling can be a complex and costly option. Although easier to implement, forced air cooling is not very effective and can introduce significant errors if used for battery model parametrisation. Existing commercially available immersion baths are not cost effective (∼£3320) and are usually too small to hold even one large pouch cell. Here, we describe an affordable but effective cooling method using immersion cooling. This bath is designed to house eight large lithium-ion pouch cells (300 mm × 350 mm), each immersed in a base oil cooling fluid (150L total volume). The total cost of this setup is only £1670. The rig is constructed using a heater, chilling unit, and a series of pumps. This immersion bath can maintain a temperature within 0.5 °C of the desired set point, it is operational within the temperature range 5–55 °C and has been validated at a temperature range of 25–45 °C.

Thermal-shock experiments for separate-effects validation of UO[formula omitted] fuel fracture models

Because of the important role that fracture plays in the behavior of ceramic UO2 fuel in a nuclear reactor environment, fracture models are a major component of fuel performance codes. As with any aspect of fuel performance, it is crucial to validate these fracture models against experimental data; however, obtaining well-controlled data for conditions representative of a reactor environment is difficult. Quenching is proposed here as a relatively simple approach for using in a laboratory environment to achieve conditions that approximate those of a reactor environment. In this paper, an experimental apparatus containing a single instrumented fuel pellet in a sealed section of copper tubing is developed and applied to a series of seven experiments in which the apparatus is first heated to a high temperature (580–680 °C) by immersion in a molten salt bath, then quenched in a cold bath (-10–4 °C). Development of these experiments was guided by numerical simulations, and post-test simulations were performed to aid in understanding the experimental behavior and assessing the accuracy of the predictions of fracture initiation and propagation. In addition, similar experiments were performed on solid copper rods to provide temperature-dependent heat transfer coefficients for use in simulations of these experiments. This study demonstrates that quenching is a viable approach for generating thermal gradients representative of those in prototypical light-water reactor conditions at powers of about 5–10 kW/m. Fracture is expected to begin at these power levels, and moderate amounts of radial and axial cracking was observed in these quenching tests.

A Cationic Contrast Agent in X-ray Imaging of Articular Cartilage: Pre-Clinical Evaluation of Diffusion and Attenuation Properties.

The aim of this study was the preliminary assessment of a new cationic contrast agent, the CA4+, via the analysis of spatial distribution in cartilage of ex vivo bovine samples, at micrometer and millimeter scale. Osteochondral plugs (n = 18) extracted from bovine stifle joints (n = 2) were immersed in CA4+ solution up to 26 h. Planar images were acquired at different time points, using a microCT apparatus. The CA4+ distribution in cartilage and saturation time were evaluated. Tibial plates from bovine stifle joints (n = 3) were imaged with CT, before and after 24 h-CA4+ bath immersion, at different concentrations. Afterward, potential CA4+ washout from cartilage was investigated. From microCT acquisitions, the CA4+ distribution differentiated into three distinct layers inside the cartilage, reflecting the spatial distribution of proteoglycans. After 24 h of diffusion, the iodine concentration reached in cartilage was approximately seven times that of the CA4+ bath. The resulting saturation time was 1.9 ± 0.9 h and 2.6 ± 2.9 h for femoral and tibial samples, respectively. Analysis of clinical CT acquisitions confirmed overall contrast enhancement of cartilage after 24 h immersion, observed for each CA4+ concentration. Distinct contrast enhancement was reached in different cartilage regions, depending on tissue’s local features. Incomplete but remarkable washout of cartilage was observed. CA4+ significantly improved cartilage visualization and its qualitative analysis.

Facile Microwave Assisted Synthesis of Silver Nanostars for Ultrasensitive Detection of Biological Analytes by SERS.

We report a very simple, rapid and reproducible method for the fabrication of anisotropic silver nanostars (AgNS) that can be successfully used as highly efficient SERS substrates for different bioanalytes, even in the case of a near-infra-red (NIR) excitation laser. The nanostars have been synthesized using the chemical reduction of Ag+ ions by trisodium citrate. This is the first research reporting the synthesis of AgNS using only trisodium citrate as a reducing and stabilizing agent. The key elements of this original synthesis procedure are rapid hydrothermal synthesis of silver nanostars followed by a cooling down procedure by immersion in a water bath. The synthesis was performed in a sealed bottom flask homogenously heated and brought to a boil in a microwave oven. After 60 s, the colloidal solution was cooled down to room temperature by immersion in a water bath at 35 °C. The as-synthesized AgNS were washed by centrifugation and used for SERS analysis of test molecules (methylene blue) as well as biological analytes: pharmaceutical compounds with various Raman cross sections (doxorubicin, atenolol & metoprolol), cell lysates and amino acids (methionine & cysteine). UV-Vis absorption spectroscopy, (Scanning) Transmission Electron Microscopy ((S)TEM) and Atomic Force Microscopy (AFM) have been employed for investigating nanostars’ physical properties.

Mechanisms of chronic pain in inflammatory rheumatism: the role of descending modulation.

Persistent pain despite satisfactory disease treatment is frequent in rheumatoid arthritis (RA) and spondyloarthritis (Spa) and may result from specific changes in central pain processing. We assessed these mechanisms further by systematically comparing thermal pain thresholds and conditioned pain modulation (CPM) between patients with active RA or Spa and healthy controls. We included 50 patients with RA and 50 patients with Spa and 100 age-matched and sex-matched controls. Heat and cold pain thresholds (HPT-CPT) were measured on the dominant forearm, and CPM was assessed by applying conditioning stimuli (immersion in a cold-water bath) to one foot and the nondominant hand in 2 successive randomized sequences. Descending pain modulation was assessed as the difference in HPTs (in °C) before and after conditioning. Larger HPT differences (ie, a larger CPM effect) reflected more efficient descending inhibition. Potential associations between changes in CPM and clinical data, including disease activity, pain intensity, and psychological and functional variables, were systematically assessed. Heat pain threshold and cold pain threshold were similar in patients and controls. The mean CPM effect was significantly weaker in patients than that in controls for conditioning applied to either the foot (0.25°C ±2.57 vs 2.79°C ±2.31; P < 0.001) or the nondominant hand (0.57°C ±2.74 vs 2.68°C ±2.12; P < 0.001). The smaller CPM effect in patients was correlated with average pain intensity, but not with disease activity or other clinical characteristics, suggesting a significant pathophysiological role for changes in endogenous pain modulation in the mechanisms of chronic pain associated with inflammatory rheumatism.

Antigenic epitope screening and functional modification of mannose enhance the efficacy of largemouth bass virus subunit vaccines.

Major capsid protein (MCP) can be used as a subunit vaccine against largemouth bass virus (LMBV). However, subunit vaccines usually have low immunogenicity. Here, to identify the major immunogenicity determinant region of the MCP gene, we truncated the MCP of the LMBV gene into four parts (MCP-1, MCP-2, MCP-3 and MCP-4). Enzyme-linked immunosorbent assay (ELISA) was used to identify the antigenicity of these four truncated MCP proteins. Then, the highly antigenic truncated protein was modified with mannose and connected with functionalized single-walled carbon nanotubes (SWCNTs) as carriers. Largemouth basses were immunized by bath immersion, challenged with LMBV on the 28th day after immunization and evaluated for related immune indicators. The results indicated that the MCP-2 protein could induce a higher antibody titre than the other truncated MCP proteins. We found that the levels of immune-related genes (TNF-α, CD40, IgM, IFNγ and IL-10) in the spleen and kidney were significantly increased in the MCP-2 and MCP-2-Man groups. ELISA results showed that the antibody content in the serum increased significantly in the MCP-2 group 7 days post-vaccination and increased with days in all the vaccinated groups, with the highest observed on the 21st day. Notably, the MCP-2-Man vaccine (10mg L-1 ) showed durability of immunoprotection efficacy that could protect largemouth basses from LMBV challenge, and the immune protection rate reached 78.94%. These results suggest that MCP-2 might be the major immunogenicity determinant region of LMBV and that the mannose-modified MCP-2 vaccine can induce stronger adaptive immune responses.

Evaluating the Gassing and Swelling Behavior of Lithium-Ion Pouch Cells Using in-Situ Pressure Analysis

The goal to reduce cost-intensive and rare elements in Lithium-ion batteries promotes the use of high-energy, nickel-rich cathodes. These materials are characterized by a high specific capacity but show an increased gas evolution during initial cycling. The main sources for the gas generation are the decomposition of electrolyte solvents at both electrodes when forming the SEI and CEI, as well as the structural release of oxygen at the cathode. A pronounced gas evolution imposes a high hazard potential during operation due to high pressures within the cell. Hence, analyzing the gas volume and the gassing period is crucial to determine optimal formation protocols and degassing steps. Existing optical or mechanical gas detection methods lack the in-situ applicability in a high-throughput production and are error-prone due to the overlaying effects of gassing and swelling. This work presents a novel method to evaluate both gassing and swelling independently to determine the gassing behavior of pouch cells with volume resolutions below 100 µl. In-situ pressure analysis is performed using an expansion cell bracket that provides a constant force on the cell, thus providing realistic and reproducible formation conditions. The method is validated using the established immersion bath measurement and three different cell chemistries are quantified: NMC/Graphite, LNMO/Graphite, and NCA/Si-Graphite. Pouch cells containing LNMO and NCA exhibited a 30 and 20 times higher gas evolution volume than conventional NMC/graphite cells, respectively. As a consequence, multi-cycle formation protocols are required as gas evolution continues within the first five charging and discharging cycles. Conclusively, a procedure to adapt the formation protocol to the gassing behavior is proposed, thus reducing processing time and ensuring the ideal degassing point.

Color of extra virgin olive oils enriched with carotenoids from microalgae: influence of ultraviolet exposure and heating

A carotenoid-rich extract containing 2.5 mg/mL of lutein and 3.3 mg/mL of β-carotene from the microalga Scenedesmus almeriensis was added to ten extra virgin olive oils from four Spanish cultivars with differing degrees of ripeness, obtaining carotenoid enriched oils with lutein and β-carotene concentrations of 0.082 and 0.11 mg/mL, respectively. Extra virgin olive oils enriched with carotenoids from microalgae were studied by analyzing the effect on color of three different treatments: ultraviolet exposure, microwave heating and immersion bath heating. The methodology was designed to simulate, in controlled laboratory conditions, the effects of household treatments. Spectrophotometric color measurements were then performed to monitor color changes in the enriched and non-enriched extra virgin olive oil samples. Enriched oils are much more chromatic, darker and redder than natural oils. After 55 days UV irradiation, 40 min microwave heating, and 72 hours thermostatic heating, the average color differences for natural/enriched extra virgin olive oils were 98/117, 15/9 and 57/28 CIELAB units, respectively. In general, increasing temperature and ultraviolet exposure produced higher CIELAB color differences in the non-enriched samples. The addition of microalga extracts to extra virgin olive oils was found to induce some color stability and may constitute a future way of increasing the daily intake of beneficial bioactive compounds such as carotenoids.

Protective Effect of Hedyotis diffusa Willd. Ethanol Extract on Isoniazid-Induced Liver Injury in the Zebrafish Model

ObjectiveThis study aims to investigate the hepatoprotective effect and molecular mechanism of Hedyotis diffusa Willd. ethanol extract (HDWE) against isoniazid (INH)-induced liver injury in the zebrafish model.MethodsINH-induced liver injury model was established by adding an immersion bath of INH in 3 days post-fertilisation (dpf) healthy transgenic zebrafish with liver-specific fluorescence (L-FABP: EGFP). HDWE and INH were given to the zebrafish to observe liver morphology and pathology, fluorescence intensity, and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD), as well as the content of glutathione (GSH). The chemical composition of HDWE was analysed using high-performance liquid chromatography coupled with a quadrupole-time-of flight hybrid mass spectrometer (HPLC-Q-TOF-MS). The bioactive compounds, molecular targets and signalling pathways of HDWE were predicted using network pharmacology. Subsequently, molecular docking was adopted to analyze the affinities between the bioactive components and targets by Autodock. Finally, in vitro experiments were conducted to further verify the findings.ResultsOur findings showed that HDWE had a remarkable protective effect on INH-induced liver injury in zebrafish. Twenty compounds in HDWE were identified. Nineteen hub targets were identified as possible targets of HDWE, and a compound-target-pathway network was constructed. Nine bioactive compounds, ten molecular targets, and seven key signalling pathways were found to play a pivotal role in the hepatoprotective effect of HDWE against INH-induced liver injury. In vitro studies revealed that the important bioactive compound quercetin-3-O-sambubioside (QSA) could significantly reverse INH-induced cell viability decreases and had a significant effect on the associated targets predicted by network pharmacology and molecular docking.ConclusionIn this study, through the research of hepatoprotective effect of HDWE and bioinformatics analysis, the bioactive compounds, important pathways and key molecular targets were discovered. These findings could provide scientific evidence for the use of HDW in liver injury and prove to help explore its efficacy and the mechanism of action.

Virulence difference between red sea bream iridovirus mixed subtype I/II and subtype II and the expression of viral and apoptosis-related genes in infected rock bream (Oplegnathus fasciatus)

In this study, the virulence of the red sea bream iridovirus (RSIV) subtype II (17RbGs isolate) and a novel RSIV mixed subtype I/II (17SbTy isolate), which was genetically characterized in a previous study, were compared. The infectivity to rock bream (Oplegnathus fasciatus) determined by infectious dose (ID50) revealed that 17RbGs isolate was significantly more infective than 17SbTy isolate using both intraperitoneal injection and bath immersion. In a cohabitation challenge test that mimicked natural conditions, the cumulative mortality of the donor (RSIV-injected rock bream) and the recipient (cohabited naïve rock bream) was significantly higher in the 17RbGs group than in the 17SbTy group, regardless of RSIV injected doses, supporting the correlation between genetic mutation and pathogenicity. In addition, the maximum viral shedding ratio identified from RSIV-infected rock bream suggested that viral transmission through infection with the 17SbTy isolate could have a lower relative risk than that of infection with the 17RbGs isolate. In particular, the odds ratio based on the spleen index after 17RbGs infection was 55.00, which was inconsistent with that of 17SbTy infection (19.38), hence supporting the virulence difference between RSIVs. Furthermore, the expression of viral genes, including DNA membrane and myristoylated protein genes with insertion and deletion mutations, and that of caspase-8, which is related to caspase-dependent apoptosis induced by RSIV infection, were significantly upregulated at 11 days post 17RbGs-infection compared to that following 17SbTy infection. Notably, although viral genes were highly expressed in the early infection stage and caspase-8 was upregulated, the low caspase-3 expression may have inhibited apoptosis, reflecting the difference in virulence between different RSIV isolates. Several virulence factors, including pathogenicity, viral shedding ratio, odds ratio, and gene expression, support that RSIV mixed subtype I/II may be a less pathogenic RSIV isolate compared with general RSIV subtype II in a natural environment.

Effects of caffeine ingestion and thermotherapy on blood orexin circulation in humans.

Caffeine and orexin can affect awakening, neuroendocrine, and sympathetic nerve function. Our previous study has reported that caffeine intake can increase human body temperature. However, little is known about the combined effects of thermotherapy and caffeine intake on human serum orexin concentrations. Forty-two healthy male subjects with age of 26.72 ± 5.05 years, height of 174.10 ± 7.09 cm, and body weight of 74.68 ± 8.91 kg participated in this study. They were randomly assigned to a control (CON) group (n = 21) and a caffeine (CAFF) group (n = 21). After thermotherapy (half-body immersion in a hot water bath at 42 ± 0.5 °C, circulating orexin level increased more (p < 0.05) in the CAFF group than in the CON group. Positive relationships between mean body temperature and orexin were observed before and after heat stimulation (p < 0.001). Caffeine intake boosted the upregulation of serum orexin concentrations in subjects undergoing thermotherapy.

A Preliminary Study of SARS-CoV-2's Permanence and Potential Infective Capacity in Mineromedicinal Waters of Copahue, Neuquén, Argentina.

Copahue Thermal Center is characterized by the presence of mineromedicinal acidic waters with high temperatures, therapeutic peloids, and relevant consortia of extremophiles species, distributed in small natural pools which cannot be disinfected. The objective of this research was to investigate the survival of SARS-CoV-2 in Copahue’s waters and its remaining infective capacity. In a first assay, a decrease of more than 50% of the initially viral load compared to the initially inoculated positive sample was detected for all the water samples analyzed. After that, two of the Copahue springs, which are used as an immersion bath in closed environments without going through any disinfection treatment, was selected to determine the viral viability. VERO cell infections were performed, with no cytopathic effect detected, but a strikingly high resistance of the virus, detecting its genome by real time PCR, during the seven days of study under laboratory conditions. SARS-CoV-2 survival in acid media was reaffirmed, which is a peculiarity for a covered virus. A decrease in the detectable viral load of the positive sample was found as the infection time passed, becoming completely negative in the subsequent blind passages. More research is needed to further study the feasibility of SARS-CoV-2 in mineromedicinal waters, especially natural acidic waters that cannot disinfected, in order to expand information about the risk to populations that are exposed to them.

Comparison of two formulations of alfaxalone for immersion anaesthesia in laboratory zebrafish (Daniorerio)

ObjectiveTo compare two commercial formulations of alfaxalone for immersion anaesthesia in laboratory zebrafish. Study designProspective, blinded, randomized study. AnimalsA total of 20 adult Danio rerio (Tuebingen strain). MethodsZebrafish were divided into two groups of 10 (five female, five male) and placed in individual immersion baths containing 10 mg L–1 of unpreserved alfaxalone (group 1) or preserved alfaxalone (group 2). Anaesthetists blinded to treatment used a composite score scale (CSS) (range 0–12) to assess fish every 30 seconds until induction of anaesthesia. Anaesthetic induction occurred when equilibrium and response to stimulus were lost. Fish were then placed in a clean water bath and scored every 60 seconds. Recovery from anaesthesia was defined as a CSS of ≤ 1. Time variables recorded were anaesthetic induction time (AIT), anaesthetic recovery time (ART) and total procedure time (TPT). Fish were observed for evidence of roupgross external pathology during the procedure. Following anaesthesia, four fish from each group were randomly chosen and euthanized for gill histopathology analysis immediately after recovery criteria were met. Data are presented as mean ± standard deviation. An independent t test was used to compare the difference in average anaesthetic time variables between groups (α = 0.05). ResultsThere were no statistical differences between groups in reported variables. TPT, AIT and ART were 10.2 ± 1.2, 1.9 ± 0.9 and 8.3 ± 1.2 minutes for group 1 and 10.8 ± 2.9, 2.4 ± 1.2 and 8.4 ± 2.7 minutes for group 2. No gross external pathology was evident, and no fish died during the experimental period. Histopathology showed normal gill pathology and no difference between the groups. Conclusions and clinical relevanceImmersion anaesthesia using 10 mg L–1 of either formulation of alfaxalone resulted in anaesthesia of similar quality and duration.

Development of a hyper-adhesive and attenuated Edwardsiella ictaluri strain as a novel immersion vaccine candidate in yellow catfish (Pelteobagrus fulvidraco)

Edwardsiella ictaluri, a Gram-negative intracellular pathogen, is the causative agent of enteric septicemia in channel catfish, and catfish aquaculture in China suffers heavy economic losses due to E. ictaluri infection. Vaccination is an effective control measure for this disease. In this study, an attenuated E. ictaluri strain was acquired through deletion mutation of the T3SS protein eseJei, and the ΔeseJei strain fails to replicate in the epithelioma papillosum of carp cells. The type 1 fimbria plays a pivotal role in the adhesion of E. ictaluri, and it was found in this study that deletion of −245 to −50 nt upstream of fimA increases its adhesion to around five times that of the WT strain. A hyper-adhesive and highly attenuated double mutant (ΔeseJeiΔfimA-245--50 strain) was constructed, and it was used as a vaccine candidate in yellow catfish via bath immersion at a dosage of 1 × 105 CFU/mL. It was found that this vaccine candidate can stimulate protection when challenged with E. ictaluri HSN-1 at 5 × 107 CFU/mL (∼20 × LD50). The survival rate was 83.61% for the vaccinated group and 33.33% for the sham-vaccinated group. The RPS (relative percent of survival) of the vaccination trial reached 75.41%. In conclusion, the ΔeseJeiΔfimA-245--50 strain developed in this study can be used as a vaccine candidate. It excels in terms of ease of delivery (via bath immersion) and is highly efficient in stimulating protection against E. ictaluri infection.

The effect of dietary supplementation of Lactobacillus rhamnosus GCC-3 fermentation product on gut and liver health, and resistance against bacterial infection of the genetically improved farmed tilapia (GIFT, Oreochromis niloticus)

High-fat diet can promote the growth of fish and save protein and its application is becoming more and more common in aquaculture. However, high-fat feed causes negative effects on fish health. The purpose of this study was to evaluate the effects of Lactobacillus rhamnosus GCC-3 fermentation product on gut and liver health and resistance against bacterial infection in genetically improved farmed tilapia (GIFT, Oreochromis niloticus) fed high fat diet. A total of 120 tilapia were randomly divided into three groups: control group, 1% GCC-3 group and 2% GCC-3 group (basal diet supplemented with 0, 10 and 20 g GCC-3/kg feed respectively). After 7 weeks of feeding, the growth performance and gut and liver health parameters of tilapia were evaluated. The gut microbiota and virome were detected by sequencing. Fish were also challenged with a combination of Aeromonas veronii and Aeromonas hydrophila by bath immersion. GCC-3 had no effect on growth performance. For gut health, the expression of HIF-1α was significantly increased in 1% GCC-3 group (P < 0.05). The expression of intestinal IL-1β, IL-6 and TNFα was significantly down-regulated in 1% and 2% GCC-3 groups (P < 0.05), and the intestinal expression of TGF-β was significantly increased (P < 0.05). GCC-3 also increased the intestinal expression of tight junction genes Tjp2a and Claudin. For liver health, GCC-3 significantly decreased liver TAG (P < 0.05). Consistently, the hepatic expression of lipid synthesis genes SREBP1, FAS and ATGL was significantly decreased in 2% GCC-3 group, while the expression of genes related to fatty acid oxidation PPARα and CPT1a increased significantly in 1% and 2% GCC-3 groups, respectively. GCC-3 increased the abundance of Cetobacterium in the gut microbiota, and led to alteration in the virome composition. Finally, the 2% GCC-3 group significantly improved the resistance of tilapia against A. veronii and A. hydrophila infection. To sum up, our results show that the dietary addition of GCC-3 fermentation product can improve the gut and liver health as well as resistance against pathogenic bacteria of tilapia.