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To describe BC collection practices at a tertiary care university hospital in a prospective observational study and examine the association between blood volume and culture yield. Analysis of all blood cultures collected during a 6-week period in 2023 at St Olav's University Hospital, Norway. Additionally, we analysed impact of antimicrobial treatment prior to sampling, sampling department, bottle type, sampling time and method, microorganisms isolated as well as patient age and sex. Only 19.7% of BC bottles were filled with the recommended 8-10 mL of blood. The overall pathogen positivity rate of all BC bottles was 6.8%, while 13.3% of the patients had at least one positive BC bottle. Patients >60 years were twice as likely to have a positive blood culture as those aged 18-60 years (16.2% vs 8.1%). Recommended filling volumes were positively associated with recovery of microbes, whereas particularly overfilling >14 mL was associated with reduced microbial yield. Overall, bottles with no growth contained a significantly higher mean blood volume than bottles with growth. This study highlights the gap between established recommendations for BC collection and current practice at our hospital. Preanalytical optimisation of BC collection is needed to improve diagnostic yield in this seriously ill patient population. Overfilling of BC bottles was negatively associated with microbial yield and should be avoided.

The emergence and widespread distribution of insecticide resistance in Anopheles gambiae s.l. raise serious concerns regarding the long-term effectiveness of vector control tools. Assessing resistance intensity provides more operationally relevant information than diagnostic-dose bioassays alone. The phenotypic intensity of resistance to pyrethroids (permethrin and deltamethrin), bendiocarb, chlorfenapyr and clothianidin in Anopheles gambiae s.l. was evaluated along a south-north transect in Benin using WHO tube tests and CDC bottle bioassays. Mortality was recorded at diagnostic doses and at higher concentrations, in accordance with WHO guidelines. At diagnostic doses, mortality rates observed with permethrin and deltamethrin were low, ranging approximately from 5-30% to 10-45%, respectively, depending on the study sites. Even at 5 × and 10 × concentrations, pyrethroid-induced mortality often remained below 90%, indicating high-intensity resistance. Resistance levels were consistently higher for permethrin than for deltamethrin, in both WHO tube tests and CDC bottle bioassays. In contrast, bendiocarb induced high mortality rates, generally exceeding 90-98% in most localities, with low to moderate resistance detected in only a few sites. Complete susceptibility of Anopheles gambiae s.l. was observed to chlorfenapyr and clothianidin, with mortality rates ≥ 98% across all communes. The intense and widespread pyrethroid resistance observed in Benin represents a major threat to interventions relying on this insecticide class. WHO tube tests and CDC bottle bioassays showed similar qualitative trends, although differences in mortality levels were observed between the two methods. The relatively preserved susceptibility to bendiocarb, as well as to chlorfenapyr and clothianidin, supports their potential use in insecticide rotation strategies, including bi-treated insecticidal nets (e.g., Interceptor® G2, PermaNet® Dual, Yorkool® G5, and Mont Inari) as well as indoor residual spraying campaigns using bendiocarb or clothianidin.

Despite known confounds of social isolation on rodent behavior, many investigators rely on singly housing mice to assess alcohol preference in preclinical models of alcohol use disorder (AUD). This protocol describes a social alcohol drinking task that allows for high-throughput, automated assessment of alcohol drinking behaviors in mice without the confounding impacts of stress induced by social isolation. The IntelliCage testing system enables investigators to simultaneously assess operant alcohol drinking behavior in up to 16 same-sex mice, each uniquely identified with a subcutaneously implanted radiofrequency identification (RFID) transponder. Access to sipper bottles is software-controlled and can be adjusted individually for each mouse. In this chronic, intermittent alcohol drinking paradigm, mice voluntarily engage in operant nosepokes to access sipper bottles containing 20% alcohol for six consecutive weeks. Alcohol bottles are only accessible every other day for 24 h at a time and can be optionally adulterated with quinine to assess punishment-insensitive drinking. Visits to each operant corner, nosepokes, and lick data are automatically recorded for each mouse. Detailed experimental methods and the open-source code to analyze the recorded data will allow investigators to assess alcohol drinking behaviors relevant to AUD in an increasingly efficient and ethologically relevant manner.

The goal of this study was to determine the effect of administering arachidonic acid (ARA) with or without the yeast Saccharomyces cerevisiae, on voluntary ethanol consumption by mice. Ethanol consumption was measured using a continuous access two-bottle choice experiment with water and 15% ethanol (vol/vol). Ethanol naïve C57BL/6J mice were orally inoculated with ARA (or acidic PBS as the control), S. cerevisiae, or both each evening for three evenings. Ethanol consumption and preference were measured on the following days. In additional experiments, male and female mice underwent chronic intermittent exposure to ethanol vapor. After a 2-week exposure protocol, these mice were given access to 15% ethanol (vol/vol) via a two-bottle choice and inoculated nightly with ARA and S. cerevisiae (or acidic PBS and mock as the control). Administration of ARA initially reduced ethanol preference and consumption by ethanol naïve female mice but the effects waned. In contrast, administration of ARA together with a fungus had a sustained effect that reduced preference and consumption. The effects of ARA and fungi were statistically significantly different from ARA alone on Day 3 of ethanol access. Chronically exposed mice that showed low initial consumption of ethanol in the two-bottle choice maintained low consumption when administered ARA and S. cerevisiae. These findings support the future development of a therapeutic regimen utilizing ARA and yeast, or targeting eicosanoids, that could be developed as a supplement to current approaches to reduce ethanol consumption.

Xylazine is a pervasive adulterant in clandestine opioid supplies. This trend is troubling, as xylazine carries its own acute side effects and its long-term cognitive and motivational effects are not known. Thus, we developed and validated a model of oral xylazine self-administration that is conducive to polysubstance studies and can easily be implemented in animal research labs. Mice underwent 4-hour drinking sessions where their only source of drinking water was adulterated with xylazine (10-1000µg/mL). Oral bioavailability and brain penetrance were validated using mass spectrometry on brain and plasma samples collected after a drinking session. Male and female mice decreased fluid intake at high concentrations of xylazine, with male mice consuming less than female mice at intermediate concentrations (100 and 300µg/mL). Female mice had decreased locomotion following drinking sessions at 100, 300, and 1000µg/mL. After a 4-hour drinking session, all mice received brain xylazine concentrations that are above the Ki (affinity) and EC50 (potency) of several known binding targets of xylazine. We then performed a three-bottle choice experiment where mice had the option of drinking from water with xylazine, water with fentanyl, or plain water. In three-bottle choice, mice consumed less xylazine-containing water than fentanyl-containing or plain water. Our results indicate that xylazine is orally bioavailable in mice, that mice will readily drink xylazine to pharmacologically and behaviorally relevant doses, and that mice prefer consuming fentanyl and water over xylazine.

Bisphenol analogues (BPs) have raised increasing concerns regarding their occurrence and risks in drinking water systems. This study systematically investigated the occurrence, spatial distribution, ecological risks, and human exposure of 14 BPs in surface water, tap water, and commercially available bottled water in Sichuan Province, China. Thirteen to fourteen BPs were detected across the samples, with sum BP concentrations (ΣBPs) ranging from 2.29 to 98.3 ng L-1 in surface water, 10.5 to 670 ng L-1 in tap water, and 2.73 to 38.2 ng L-1 in bottled water. Relatively higher accumulation of ΣBPs was found in surface water from the Northeast Sichuan Economic Zone and the Chengdu Plain Economic Zone (median: 13.0 and 11.1 ng L-1, respectively), compared to the Panxi Economic Zone (median: 4.27 ng L-1). Ecological risk assessment indicated that BPA, BPBP, and BPG may pose low to moderate risks to ecosystems, while the estrogenic risk estimated using the estrogen equivalent was negligible. Furthermore, the estimated daily intake (EDI) of BPA from tap water exceeded the tolerable daily intake recommended by the European Food Safety Authority, indicating a potential health risk from dietary exposure.

Rapid and selective detection of Gram-negative bacteria is important for clinical diagnosis and for monitoring environmental and food-related samples. In this study, we developed a label-free ISFET sensor based on a p-type silicon/SiO2 platform modified with ZnO and colistin for the detection of Gram-negative bacteria. The sensor was evaluated against several Gram-negative species, including Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Salmonella enterica serovar Enteritidis, Klebsiella oxytoca, and Klebsiella pneumoniae, with the Gram-positive species Staphylococcus aureus and Enterococcus faecium used as negative controls. The device showed measurable changes in drain-source current upon bacterial binding, and the standard curves indicated a low detection limit and a linear response within the tested concentration range. The calculated limits of detection were approximately 137 CFU/mL for E. coli, 169 CFU/mL for A. baumannii, 188 CFU/mL for S. enterica, 218 CFU/mL for K. oxytoca, 235 CFU/mL for K. pneumoniae, and 289 CFU/mL for P. aeruginosa. The sensor stability was also assessed over 14 days in the E. coli assay. In addition, real-sample experiments in commercial bottled drinking water and human urine spiked with E. coli showed good agreement with PBS measurements, with a recovery of 101.13% ± 3.08% and an RSD of 3.05% in bottled water. These results indicate that the proposed platform has potential for selective detection of Gram-negative bacteria under controlled laboratory conditions and in tested real matrices.

It is becoming increasingly clear that chronic exposure to lower levels of ethanol impact learning and behavior. To determine the impact of chronic low-dose ethanol exposure on sensitivity to changes in stimulus value, a conditioned taste aversion procedure was used. Adult male and female mice underwent a sucrose two bottle-choice drinking paradigm. Each day, mice received an injection of either low-dose ethanol (0.5g/kg) or saline two hours after sucrose access for 20 days. This was followed by a lithium chloride (LiCl)-induced conditioned taste aversion (CTA) paradigm in which 0.15M LiCl or vehicle injection was administered immediately after sucrose consumption for three days. On the fourth day, changes in sucrose consumption were analyzed. Chronic exposure to low-dose ethanol did not affect sucrose consumption in either female of male mice during two-bottle choice. In female mice, a history of chronic low-dose ethanol exposure blocked the development of LiCl-induced CTA. A history of chronic low-dose ethanol did not impact LiCl-induced CTA in male mice as both ethanol-naïve and -exposed male mice who underwent LiCl pairing reduced sucrose consumption. This suggests that low-dose ethanol alters aversion-related learning in female mice which may have implication for development of aberrant behavior and risk for alcohol use disorder (AUD).

The micro/nanoplastics (MNPs) have been evidenced to exert detrimental effects on the blood-brain barrier (BBB) and the central nervous system (CNS). However, there is still a lack of effective research models on the mechanism of nerve injury caused by microplastics particles. This study focuses on analyzing the particle size characteristics of MNPs precipitated from plastic water bottles under different conditions of storage and uses 3D BBB microfluidic chips to assess the permeability and dynamic neurotoxicity of MNPs. The results showed that there was a significant increase in the average diameter of MNPs in purified water stored in plastic bottles. Moreover, the cultivation of BBB cells or neuronal cells with two different particle sizes of MNPs showed a significant decrease in cell survival rates. When MNPs were infused into the peripheral unit of the biomimetic chip, they could penetrate from the endothelial cell unit to the neuronal unit and induce a dynamic injury process with neuroinflammation, accompanied by tight junction disruptions, increased ROS levels, decreased mitochondrial membrane potential, decreased lipid droplet levels, and increased inflammatory effects. The research results based on engineering 3D microfluidic chips lay the foundation for a deeper understanding of the inflammatory damage to nerve cells caused by MNPs crossing the BBB.