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Despite the general classification of L. monocytogenes strains as equally virulent by global safety authorities, molecular epidemiology reveals diverse subtypes in food, processing environments, and clinical cases. This study focuses on a highly virulent strain associated with a listeriosis outbreak in Italy in 2022, providing insights through comprehensive foodomics approaches, with a specific emphasis on quantitative proteomics. In particular, the ST155 strain of L. monocytogenes strain was subjected in vitro to growth stress conditions (NaCl 2.4 %, pH 6.2, T 12 °C), mimicking the conditions present in the frankfurter, its original source. Then, the protein expression patterns were compared with those obtained in optimal growth conditions. Through quantitative proteomic analysis and bioinformatic assessment, different proteins associated with virulence during the exponential growth phase were identified. This study unveils unique proteins specific to each environment, providing insights into how L. monocytogenes adapts to conditions that are similar to those encountered in frankfurters. This investigation contributes valuable insights into the adaptive strategies of L. monocytogenes under stressful conditions, with implications for enhancing food safety practices., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Vibrio diabolicus is widely distributed in the marine environment and is an important pathogen of aquatic organisms such as shrimp, fish, and mollusks. The emergence of multi-drug resistance among these bacteria has resulted in a global public health problem, which requires alternative treatment approaches, such as phage therapy. In the present study, we isolated the phage vB_Vc_SrVc2 from the hepatopancreas of white shrimp showing symptoms of acute hepatopancreatic necrosis disease (AHPND) and evaluated the efficacy of this phage in preventing the mortality associated with V. diabolicus presence in shrimp culture. Phage vB_Vc_SrVc2 belongs to the genus Maculvirus within the family Autographiviridae and demonstrates high lytic ability against Vibrio isolates. It exhibits substantial resilience across a broad range of temperatures, salinity levels, UV radiation and chloroform exposure. The phage genome size is 43,157 bp, with a GC content of 49.2%, that encodes 49 putative ORFs, and no tRNAs. Compared to phage vB_Vc_SrVc9, it shows three single nucleotide polymorphisms, two small deletions, and one nucleotide insertion, which result in slightly different infectivity profiles. No lysogeny-related genes were detected in the vB_Vc_SrVc2 genome. In vivo trials revealed that the phage offered protection against V. diabolicus infection by delaying mortality onset by at least 40 h post-infection (hpi). Although phage vB_Vc_SrVc2 alone significantly reduced mortality associated with V. diabolicus infection, this capacity could be enhanced with multiple doses and/or the use of phage cocktails or probiotics. Overall, phage vB_Vc_SrVc2 shows promising potential for therapeutic application in the aquaculture industry against V. diabolicus infections., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Listeria monocytogenes was the etiologic agent in nearly all recent outbreaks in North America attributed to pasteurized dairy products, whereas Escherichia coli O157 infections were responsible for most of the rare, yet serious complications from outbreaks involving unpasteurized dairy. This study determined the susceptibility of selected strains of L. monocytogenes and Shiga toxin-producing E. coli (STEC) to commercial phage preparations and their ability to control these pathogens in pasteurized and raw milk during 7-day storage at 7 °C. Both phage products demonstrated high lytic efficiency against 17 strains of L. monocytogenes whereas the efficiency of E. coli phages was more variable against 11 strains of O157 and non-O157 STEC. Broth microdilution assays identified effective endpoint multiplicities of infection (MOI) ranging from log 2.53 to 5.13, which differed between strains of L. monocytogenes and phage products. Mean log MOIs of E. coli phages against STEC also varied within and between products from 0.43 to 7.05. Despite these observations, the change in counts over time of three L. monocytogenes strains exposed to phage in pasteurized milk (log MOI 6) was similar with counts ∼ 4 log CFU/mL lower than control at day 7. Results for STEC O157 varied by strain but counts were lower than control in all cases by 72 h thorough day 7. Titers on isolates of both pathogens isolated from pasteurized milk indicated that the surviving populations were less susceptible to phage. The addition of a phage preparation to raw milk did not reduce populations of either pathogen or affect the change in counts of any strain over time when compared to control. Reduced efficacy in raw milk may be attributed to reduced phage binding as titers in raw milk decreased steadily (∼2 log PFU/mL) during storage. Commercial phage products may be a promising pathogen control intervention for pasteurized dairy products, warranting further investigation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dennis D'Amico reports financial support was provided by National Institute of Food and Agriculture and The George Walker Milk Research Fund. Products used in this research were provided free of charge by Intralytix and Micreos food Safety- DD If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A novel video-based real-time system based on AI (artificial intelligence) was developed to detect clinical signs in fish exposed to pathogens. We selected a White Spot Disease model involving rainbow trout as the experimental animal and the parasitic ciliate Ichthyophthirius multifiliis as a pathogen. We compared two identical fish tank systems: one tank was infected by co-habitation, whereas the other tank was kept non-infected (sham infection). The two fish tanks were separately video monitored (full top and side view) during the course of infection, during which fish were removed whenever they developed clinical signs (direct visual inspection by the observer). Image analysis (object detection, classification and tracking) was used to track behavioural changes in fish (in every recorded video frame), focusing on movement patterns and spatial localisation. Initially, the two fish groups (infected and non-infected) exhibited similar behaviour and non-infected fish did not change behaviour during the 15 d observation period (from 5 d before infection until 10 dpi). At 4, 7, 8, 9 and 10 dpi some infected fish showed clinical signs (equilibrium disturbance, gasping and lethargy) and were removed from the experiment. Anorexia occurred from 5 dpi and a gradual progression of gasping behaviour was noted, whereas the frequency of fish flashing (rubbing/scratching against objects) was low. Equilibrium disturbances and the development of white spots in the skin appeared to be a much later (8-10 dpi at this temperature) indicator of infection. The video analysis showed a general distribution of non-infected fish in all parts of the fish tank during the entire experiment, whereas infected fish already at 4-5 dpi moved towards higher water currents in the top and bottom positions. This change of fish positioning within the tank appeared as a promising early indicator of infection. The study suggests that continuous monitoring of fish behaviour using AI can potentially optimise the timing of humane endpoints, indicate disease signs earlier and thereby improve animal welfare in both animal experimentation and in aquaculture settings., (© 2024 The Author(s). Journal of Fish Diseases published by John Wiley & Sons Ltd.)

24 h cold exposure (4°C) is sufficient to reduce pathogen susceptibility in Arabidopsis thaliana against the virulent Pseudomonas syringae pv. tomato ( Pst ) strain even when the infection occurs five days later. This priming effect is independent of the immune regulator Enhanced Disease Susceptibility 1 (EDS1) and can be observed in the immune-compromised eds1-2 null mutant. In contrast, cold priming-reduced Pst susceptibility is strongly impaired in knock-out lines of the stromal and thylakoid ascorbate peroxidases (sAPX/tAPX) highlighting their relevance for abiotic stress-related increased immune resilience. Here, we extended our analysis by generating an eds1 sapx double mutant. eds1 sapx showed eds1 -like resistance and susceptibility phenotypes against Pst strains containing the effectors avrRPM1 and avrRPS4. In comparison to eds1-2 , susceptibility against the wildtype Pst strain was constitutively enhanced in eds1 sapx . Although a prior cold priming exposure resulted in reduced Pst titers in eds1-2 , it did not alter Pst resistance in eds1 sapx . This demonstrates that the genetic sAPX requirement for cold priming of basal plant immunity applies also to an eds1 null mutant background.

Background: Respiratory infection is a major cause of acute exacerbation of chronic obstructive pulmonary disease (AECOPD). We investigated the presence of bacterial and viral pathogens and clinical features in patients with AECOPD., Methods: This retrospective study included 1,186 patients diagnosed with AECOPD from 28 hospitals in South Korea between 2015-2018. Pathogen identification rates, basic characteristics and clinical features, and associated factors for infection with potentially drug-resistant (PDR) pathogens were evaluated using microbiological tests., Results: Bacteria, viruses, and both were found in 262 (22.1%), 265 (22.5%), and 129 (10.9%) patients, respectively. The most common pathogens were Pseudomonas aeruginosa (17.8%), Mycoplasma pneumoniae (11.2%), Streptococcus pneumoniae (9.0%), influenza A virus (19.0%), rhinovirus (15.8%), and respiratory syncytial virus (6.4%). A history of pulmonary tuberculosis (OR 1.66; P=0.046), bronchiectasis (OR 1.99; P=0.032), and triple inhaler use within six months (OR 2.04; P=0.005) were significant associated factors for PDR pathogen infection. Hospital stay length (15.9 days vs. 12.4 days; P=0.018) and ICU admission rates (15.9% vs. 9.5%; P=0.030) were increased in patients infected with PDR pathogens., Conclusions: This study indicates that various types of pathogens are implicated during AECOPD. However, further research is needed to confirm whether these pathogens influence AECOPD development and progression.

The bacterium Legionella pneumophila secretes numerous effector proteins that manipulate endoplasmic reticulum (ER)-derived vesicles to form the Legionella-containing vacuole (LCV). Despite extensive studies, whether the LCV membrane is separate from or connected to the host ER network remains unclear. Here, we show that the smooth ER (sER) is closely associated with the LCV early in infection. Remarkably, Legionella forms a distinct rough ER (rER) niche at later stages, disconnected from the host ER network. We discover that host small GTPases Rab10 and Rab4 and an ER protein, BAP31, play crucial roles in transitioning the LCV from an sER to an rER. Additionally, we have identified a Legionella effector, Lpg1152, that binds to BAP31. Interestingly, the optimal growth of Legionella is dependent on both BAP31 and Lpg1152. These findings detail the complex interplay between host and pathogen in transforming the LCV membrane from a host-associated sER to a distinct rER., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Animals constantly face microbial challenges, and microbe-mediated infection protection is crucial for host survival. Identifying specific bacteria and their interactions with host intracellular surveillance systems is important but challenging. Here, we develop a "probiotics" screening system that identifies Escherichia coli mutants, such as ΔymcB, which protect hosts from Pseudomonas aeruginosa PA14 infection by activating the mitochondrial unfolded protein response (UPR mt ). Genetic screening reveals that MDSS-1, a neuronal transmembrane protein, is crucial for sensing ΔymcB and triggering intestinal UPR mt . MDSS-1 functions as a potential receptor in ASE neurons, detecting ΔymcB and transmitting signals through neuropeptides, GPCRs, Wnt signaling, and endopeptidase inhibitors to activate intestinal UPRmt and enhance protection. Constitutive activation of MDSS-1 in ASE neurons is sufficient to induce UPR mt and confer infection resistance. This study uncovers a neuron-intestine communication mechanism, where ASE neurons detect bacteria and modulate the intestinal mitochondrial surveillance system for host adaptation to pathogens., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Helicobacter pylori (Hp) has been postulated as an infectious trigger of psoriatic disease, namely psoriasis (Ps) and psoriatic arthritis (PsA), but meticulous antibody (ab) reactivity against all dominant and subdominant Hp antigens in demographically matched PsA and Ps patients and healthy controls has not been performed so far. IgG anti-Hp ab testing was performed by combining immunoblotting and line assays in 263 serum samples from 89 patients with PsA, 114 patients with Ps, and 60 demographically matched healthy controls (HCs). Anti-Hp positivity did not differ between PsA, Ps, and HCs (P > 0.05 for all comparisons). In PsA, anti-p75, anti-p67-FSH, anti-p66-UreB, anti-p54-flagellin, anti-p41, and anti-p30-OMP abs were more frequent in patients compared to HCs (P < 0.001, P = 0.028, P = 0.010, P = 0.003, P = 0.012, P = 0.020 respectively). In Ps, anti-p66-UreB and anti-p54-flagellin abs were more frequent than HC (P = 0.015 and P = 0.011, respectively), while anti-p50 abs were less frequent than HCs (P = 0.008). Anti-p75, anti-p67-FSH, anti-p50, anti-p41, anti-p30-OMP, anti-p29 = UreA and anti-p26 ab levels were higher in PsA compared to Ps (P = 0.012, P = 0.036, P < 0.001, P = 0.021, P = 0.002, P = 0.006 and P = 0.021 respectively). DAS28 scores were positively correlated with anti-p19 ab levels (r = 0.349, P = 0.050) in PsA patients by linear regression analysis. No other significant clinical association with anti-Hp responses was noted in patients with PsA and Ps. Our results demonstrate that several antigen-specific anti-Hp abs are more frequent in patients with psoriatic disease; however, negative correlations also exist, raising doubts about whether Hp is immunologically linked to psoriatic disease., Competing Interests: Declarations. Competing interests: Thomas Scheper and  Wolfgang Meyer are employees of Euroimmun AG. All other authors have nothing to declare., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Microbiomes are sustained through infinite yet mutually interacting microbial communities, with bacteria and fungi serving as the major constituents. In recent times, microbial interventions have become popular for microbiome manipulation to achieve sustainable goals. Whether and how the introduced biocontrol agent drives fungal microbial assemblages (mycobiome) and the role of interkingdom signaling in shaping the microbiome structure and function remain poorly understood. Here, we implemented wild-type (WT) Bacillus subtilis L1-21 and its quorum quenching (QQ) mutants (L1-21Δytnp, and L1-21Δyxel) individually and as consortia to explore the enrichment patterns of key mycobiome members in Huanglongbing (HLB) infected citrus compartments including leaf endosphere, root endosphere, and rhizosphere soil. The application of WT and its QQ mutants produced differential mycobiome enrichment across citrus compartments. Our findings reveal that application of WT B. subtilis enriched beneficial fungi such as Trichoderma (15.82 %) in leaf endosphere. In contrast, pathogenic fungi Fusarium (47.5 %) and Gibberella (0.47 %) involved in citrus root decline were adundant in the L1-21Δytnp treated root endosphere while Nigrospora (11 %) was predominant in L1-21Δyxel treated leaf endosphere, affirming the role of bacterial quorum sensing (QS) molecules in shaping the fungal community composition. In general, based on the fungal functional prediction, fungal pathogens were highly abundant in mutant-treated plants, particularly in leaf endosphere (L1-21Δytnp: 25 %; L1-21Δyxel: 36.35 %) compared to WT (20.93%). Additionally, some fungal members exhibited strong compartment specificity and both mutants induced distinct mycobiome shifts in rhizosphere soil, leaf, and root endopshere. In conclusion, B. subtilis QQ modifies bacterial QS networks facilitating beneficial fungi to establish, while loss of QQ leads to enrichment of pathogenic fungal groups. Our study provides a direct link of perception and regulation of mycobiome through bacterial-based QS and QQ system, and its association with disease outcomes., Competing Interests: Declaration of competing interest The authors have no conflicting interests related to this manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Environmental matrices have been considered of paramount importance in the spread of antibiotic-resistance; however, the role of drinking waters is still underexplored. Therefore, a scoping review was performed using a systematic approach based on PRISMA guidelines, with the aim of identifying and characterizing antibiotic-resistance in tap water, specifically, water treated at a potabilization plant and provided for drinking use through a water distribution system. The review included 45 studies, the majority of which were conducted in upper-middle-income economies (42.2%), mainly from the Western Pacific region (26.7%), followed by Europe (24.4%). Most of the papers focused on detecting antibiotic-resistant bacteria (ARB), either alone (37.8%) or in combination with antibiotic-resistant genes (ARGs) (26.7%). Multidrug-resistance profile was often identified in heterotrophic bacteria, including various species of nontuberculous mycobacteria, Pseudomonas spp., and Aeromonas spp., which were especially resistant to penicillins, cephalosporins (including 3rd-generation), and also to macrolides (erythromycin) and tetracyclines. Resistance to a wide range of antibiotics was also prevalent in fecal bacteria, e.g., the Enterobacteriaceae family, with common resistance to (fluoro)quinolones and sulfonamide groups. ARGs were investigated either in bacterial strains isolated from tap waters or directly in water samples, and the most frequently detected ARGs belonged to β-lactam, sulfonamide, and tetracycline types. Additionally, mobile genetic elements were found (i.e., int1 and tnpA). Sulfonamides and macrolides were the most frequently detected antibiotics across countries, although their concentrations were generally low (<10 ng/L) in Europe and the United States. From a health perspective, tap water hosted ARB of health concern based on the 2024 WHO bacterial priority pathogens list, mainly Enterobacteriaceae resistant to 3rd-generation cephalosporin and/or carbapenem. Despite the fact that tap water is treated to meet chemical and microbiological quality standards, current evidence suggests that it can harbor antibiotic-resistance determinants, thus supporting its potential role in environmental pathways contributing to antibiotic resistance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Stomata, the small pores on the surfaces of leaves and stems, are crucial for gas exchange in plants and also play a role in defense against pathogens. The stomatal movement is not only influenced by surrounding light conditions but also by the presence of foliar pathogens. To put it more crisply, certain light wavelengths such as blue or strong red light, cause stomatal opening, which tragically makes it easier for bacteria to enter through opened stomata and causes disease progression in plants. Illumination of blue or intense red light autophosphorylates phototropin, a blue light photoreceptor protein kinases that in turn activates signaling cascade to open stomata. Undoubtedly stomatal defense is a fascinating aspect of plant immunology, especially in plant-foliar pathogen interaction. During these interactions, stomata fundamentally serve as entry points for intrusive pathogens and initiate plant defense signaling cascade. The present review highlights how the light-activated photoreceptors like cryptochromes (CRYs), phytochromes (phys), and UV-receptors (UVRs) influence the stomatal movement and defense signaling after foliar pathogen intrusion. It also explores the link between stomatal defense, light signaling, and plant immunity, which is vital for safeguarding crops against pathogens., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Neutrophils are a part of the innate immune system and produce reactive oxygen species (ROS) to extinguish pathogens. The major source of ROS in neutrophils is NADPH oxidase, which is fueled by NADPH generated via the pentose phosphate pathway; however, it is unclear how other accessory glucose metabolism pathways and mitochondrial activity influence the respiratory burst. We examined the temporal dynamics of the respiratory burst and delineated how metabolism changes over time after neutrophil activation. Bone marrow-derived neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and the respiratory burst was measured via extracellular flux analysis. Metabolomics experiments utilizing 13 C 6 -glucose highlighted the activation of glycolysis as well as ancillary pathways of glucose metabolism in activated neutrophils. PMA stimulation acutely increased 13 C enrichment into glycerol 3-phosphate (G3P) and citrate, whereas increases in 13 C enrichment in the glycogen intermediate, UDP-hexose, and end products of the hexosamine and serine biosynthetic pathways occurred only during the late phase of the oxidative burst. Targeted inhibition of the G3P shuttle, glycogenolysis, serine biosynthesis, and mitochondrial respiration demonstrated that the G3P shuttle contributes to the general magnitude of ROS production; that glycogen contributes solely to the early respiratory burst; and that the serine biosynthetic pathway activity and Complex III-driven mitochondrial activity influence respiratory burst duration. Collectively, these results show that the neutrophil oxidative burst is highly dynamic, with coordinated changes in metabolism that control the initiation, magnitude, and duration of ROS production., Competing Interests: Conflict of Interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In introductory microbiology courses for non-biology majors, it can be difficult to ensure that students gain a functional understanding of the interplay between the host immune system and an invading infectious agent. Immune Battle is a board game that allows students to explore pathogen evolution and the immune system's response to incursions in a captivating way, leveraging battleship and worker placement styles of gameplay. Students play in teams as either a pathogenic microbe attempting to reproduce and transmit itself to new hosts or as the immune system trying to defend the host from infection. Immune Battle seeks to mimic real-world interactions between immune cells and microbes using board game mechanics. For example, students will quickly find that adaptive immune system actions are better at fighting off microbial incursions than innate immune actions but require more time to activate. In this way, Immune Battle reinforces student's understanding of the immune system and pathogenic virulence in an exciting and interactive way that keeps them thinking about host defenses long after the end of the class period or exam. This board game is well suited for advanced high school courses and introductory college-level courses that have only limited time to cover the immune system and microbiology. Documents containing the necessary components of the board game (not including a six-sided die) are included in the supplemental materials, so educators can print out their own copies and use this board game in their classes with no cost to themselves or their students., Competing Interests: The author declares no conflict of interest.

In recent years, a disease called "Baotou" has been causing large-scale yield reductions of Gracilariopsis lemaneiformis in China. Interestingly, Vibrio alginolyticus , which once reported to be a probiotic or pathogen for multiple marine organisms, was strongly proved to be the pathogen causing "Baotou" disease in this study. Analysis of 16S rRNA gene profiling revealed that V. alginolyticus was the most abundant and dominant bacterium on the algal thalli suffering from "Baotou" disease, whereas its presence was scarcely detected on healthy thalli. Scanning electron microscope (SEM) analysis revealed that a large number of V. alginolyticus cells were found to be attached to the algal thalli with "Baotou" disease and the rotten thalli acquired by the lab infection treatment. V. alginolyticus could cause the rotten symptoms which were consistent with those of "Baotou" disease. According to Koch's postulates, V. alginolyticus was identified as the pathogen causing "Baotou" disease.IMPORTANCEA highly contagious disease known as "Baotou" disease has persistently triggered significant yield reductions in G. lemaneiformis throughout China. The pathogen of "Baotou" disease was isolated and identified as V. alginolyticus in this study. Interestingly, V. alginolyticus was once reported to be a probiotic to Saccharina japonica , and pathogen to Haliotis diversicolor , Paleopneustes cristatus , Ruditapes decussatus, and Litopenaeus vannamei . The study indicates that V. alginolyticus play a significant role in the competition or co-existence between G. lemaneiformis , S. japonica , abalone, sea urchin, bivalve, and shrimp.

As natural populations continue to decline globally, direct forms of intervention are increasingly necessary to prevent extinction. One type of intervention, known as demographic rescue, occurs when individuals are added directly to a population to increase abundance and ultimately prevent population extinction. However, the role of infectious disease in demographic rescue remains unknown. To examine the effects of pathogens on demographic rescue, we used a host-pathogen system with the aquatic crustacean Daphnia dentifera as the host and the fungus Metschnikowia bicuspidata as the pathogen. We constructed a randomized 3 × 2 factorial experiment with three rescue treatments (none, low, high) and two pathogen treatments (unexposed, exposed), where the pathogen was introduced via infected individuals during rescue events. We found that adding more individuals to demographically depressed populations increased abundance over the short term; highly supplemented populations initially had 62% more individuals than populations that had no introduced individuals. However, by the end of the experiment, populations that did not have any individuals introduced averaged 640% higher abundance than populations where infected individuals had been added. Thus, the introduction of infected individuals can result in worse demographic outcomes for populations than if no rescue is attempted., (© 2024 The Author(s). Ecology published by Wiley Periodicals LLC on behalf of The Ecological Society of America.)

Polymorphonuclear lymphocytes (PMNs) are terminally differentiated phagocytes with pivotal roles in infection, inflammation, tissue injury, and resolution. PMNs can display a breadth of responses to diverse endogenous and exogenous stimuli, making understanding of these innate immune responders vital yet challenging to achieve. Here, we report a 22-color spectral flow cytometry panel to profile primary human PMNs on population and single cell levels for surface marker expression of activation, degranulation, phagocytosis, migration, chemotaxis, and interaction with fluorescently labeled cargo. We demonstrate the surface protein response of PMNs to phorbol ester stimulation compared to untreated controls in an adherent PMN model with additional analysis of intra- and inter-subject variability. PMNs challenged with the Gram-negative bacterial pathogen Neisseria gonorrhoeae revealed infectious dose-dependent changes in surface marker expression in bulk, population-level analysis. Imaging flow cytometry complemented spectral cytometry, demonstrating that fluorescence signal from labeled bacteria corresponded with bacterial burden on a per-cell basis. Spectral flow cytometry subsequently identified surface markers which varied with direct PMN-bacterium association as well as those which varied in the presence of bacteria but without phagocytosis. This spectral panel protocol highlights best practices for efficient customization and is compatible with downstream approaches such as spectral cell sorting and single-cell RNA-sequencing for applicability to diverse research questions in the field of PMN biology., Competing Interests: The authors declare no conflicts of interest.

Despite the large wild boar population in South Korea, research on infectious diseases is limited. We investigated the prevalence of infectious diseases caused by Porcine Circovirus Type 2 (PCV2), Aujeszky's Disease Virus (ADV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and Mycoplasma hyopneumoniae in 400 wild boars in South Korea. PCV2 antibodies were detected in 223 samples, antigens in 112, and both in 101. ADV and PRRSV antibodies were detected in one and three samples, respectively. M. hyopneumoniae antibodies were identified in 28 samples. PCV2 was detected in all nine regions, M. hyopneumoniae in five, PRRSV in three, and ADV in one. Infectious-disease monitoring is crucial to safeguard wild boars.

Introduction: This review aims to assess the safety and efficacy of the use of ultraviolet-C technology for disinfecting aircraft and compare it with other methods currently used in the aviation industry., Methods: The authors conducted a comprehensive, systematic review of the literature on disinfection of aircraft. Independent double reviews were conducted and consultations with a third reviewer were performed in the event of disagreements., Discussion: Although infectious disease transmission in aircraft cabins has been shown to be low, a recent study has described reports of passengers on commercial aircraft infecting other passengers. Incorporating ultraviolet-C technology into aircraft disinfection protocols holds the potential to add a significant level of risk mitigation to effectively reduce disease transmission and enhance safety. DeJohn C, Belland K, Garcia D. Methods of aircraft disinfection to reduce airborne infectious disease transmission. Aerosp Med Hum Perform. 2024; 95(12):930-936.

The World Organisation for Animal Health (WOAH) seeks to improve animal health by promoting safe trade in animals and their products, setting standards for diagnostic methods, detection of emerging diseases and confirmation of listed diseases in clinically affected animals and surveillance in healthy populations. WOAH launched the Aquatic Animal Health Strategy (AAHS) in 2021 in recognition of the growing importance of aquatic animal products in global food security. Disease is currently a major limiting factor in the sustainable growth and development of the aquaculture industry, impacting the industry's ability to increase yields to meet future food demands. A key aim of the AAHS is to ensure that scientifically sound standards are set to manage risks and facilitate safe trade, improve aquatic animal health and welfare, build capacity to strengthen aquatic animal health services provided by WOAH and ensure that responses to aquatic animal health issues are coordinated and timely, at both regional and global level. This article evaluates some of the latest scientific and policy advances, as well as obstacles for the implementation of the AAHS.

This study investigated the diversity of thermophilic Campylobacter species isolated from three New Zealand freshwater catchments affected by pastoral and urban activities. Utilising matrix-assisted laser desorption ionisation-time of flight and whole genome sequence analysis, the study identified Campylobacter jejuni (n = 46, 46.0%), C. coli (n = 39, 39%), C. lari (n = 4, 4.0%), and two novel Campylobacter species lineages (n = 11, 11%). Core genome sequence analysis provided evidence of prolonged persistence or continuous faecal shedding of closely related strains. The C. jejuni isolates displayed distinct sequence types (STs) associated with human, ruminant, and environmental sources, whereas the C. coli STs included waterborne ST3302 and ST7774. Recombination events affecting loci implicated in human pathogenesis and environmental persistence were observed, particularly in the cdtABC operon (encoding the cytolethal distending toxin) of non-human C. jejuni STs. A low diversity of antimicrobial resistance genes (aadE-Cc in C. coli), with genotype/phenotype concordance for tetracycline resistance (tetO) in three ST177 isolates, was noted. The data suggest the existence of two types of naturalised waterborne Campylobacter: environmentally persistent strains originating from waterbirds and new environmental species not linked to human campylobacteriosis. Identifying and understanding naturalised Campylobacter species is crucial for accurate waterborne public health risk assessments and the effective allocation of resources for water quality management., (© 2024 The Author(s). Environmental Microbiology published by John Wiley & Sons Ltd.)

To study the role of intestinal flora in the development of bloodstream infections (BSIs). 42 patients and 19 healthy controls (HCs) were screened into the study and their intestinal flora was measured by 16S rRNA gene sequencing. The bacterial diversity was significantly lower in the BSI group compared with that in the HCs (P < 0.001), and beta diversity was significantly differentiated between the two groups (PERMANOVA, P = 0.001). The four keystone species [Roseburia, Faecalibacterium, Prevotella, and Enterococcus (LDA > 4)] differed significantly between the two groups. Dysbiosis of fecal microbial ecology is a common condition present in patients with BSI. The proliferation of certain pathogens or reduction of SCFA-producing bacteria would cause susceptibility to BSI., Competing Interests: Declarations. Conflict of Interest: The authors declare that they have no competing interests. Ethical and Consent: The study was approved by the Institutional Ethics Board of Jieyang People’s Hospital, including all procedures (Ethics approval NO.2022028). With the approval of the Ethics Committee, medical records and biospecimens obtained from previous clinical diagnoses and treatments may apply for a waiver of informed consent when used anonymously for research and publication., (© 2024. The Author(s).)

This is the first study to assess the impact of substitution of shortening (50%) with sunflower-beeswax oleogel in cake formulations on the inactivation kinetics of Salmonella spp. and the quality attributes of cakes baked in conventional (CO) and microwave (MWO) ovens. Four distinct cake samples were examined in the study: Cake samples containing oleogel prepared in two different ovens (Oleo-Cake-CO and Oleo-Cake-MWO) and control cake samples baked in two different ovens (Cont-Cake-CO and Cont-Cake-MWO). The control-batter and oleogel-batter demonstrated shear thinning behavior (pseudoplastic, n < 1), with a good fit to the Power Law model, but viscosity and viscoelastic moduli decreased when the oleogel was used in place of shortening in cake recipes. In addition, the Cont-Cake-MWO had the greatest special volume value (2.31 ± 0.04 mL/g), whereas the Oleo-Cake-CO had the lowest (1.65 ± 0.02 mL/g) (p < 0.05). Furthermore, compared to the samples baked with CO, lower water activity and moisture values (p < 0.05) were observed in the samples baked in MWO due to their higher cooking loss values (p < 0.05). In all baking techniques, the addition of oleogel to the cake formula, used as a fat substitute, resulted in higher values for cohesiveness, hardness, springiness, and chewiness (p < 0.05). As a result, the inactivation of Salmonella in cakes slightly reduced with using oleogel as a fat substitute (p > 0.05), whereas it affected some quality properties of cakes baked with both heat treatments., (© 2024 The Author(s). Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Bats are increasingly in the focus of disease surveillance studies as they harbor pathogens that can cause severe human disease. In other host groups, ectoparasitic arthropods play an important role in transmitting pathogens to humans. Nevertheless, we currently know little about the role of bat-associated ectoparasites in pathogen transmission, not only between bats but also to humans and other species, even though some of these parasites occasionally feed on humans and harbor potentially zoonotic organisms. In this work, we summarize current knowledge on the zoonotic risks linked to bat-associated ectoparasites and provide novel risk assessment guidelines to improve targeted surveillance efforts. Additionally, we suggest research directions to help adjust surveillance strategies and to better understand the eco-epidemiological role of these parasites., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Derivatives of chitosan-ethyl formate polymers (Chs-EF) show promise as biologically relevant materials. The novelty of this study lies in the innovative use of Chs-EF doped with zinc oxide nanoparticles and beta-cyclodextrin, which significantly enhances the polymers' protective activities against Alternaria early blight disease in Vicia faba by improving both disease resistance and plant health. After doping Chs-EF with zinc oxide nanoparticles (ZnONPs) and inserting it into the beta-cyclodextrin (CD), two products emerged: Chs-EF/ZnONPs and Chs-EF/CD. Using βCD and ZnONPs to modify the Chs-EF polymer improves the optical properties of the generated polymers. Also, the energy gab values of the modified polymers (Chs-EF/ZnONPs and Chs-EF/βCD) were 3.3 and 3.7 eV, respectively, while energy gab value of the Chs-EF polymer was 3.9 eV. In this study, the effects of ZnONPs, chitosan, β-CD, and Chs-EF/ZnONPs on Alternaria solani early blight disease in Vicia faba plants were investigated. The treatments were evaluated based on disease symptoms and a disease index (DI) to assess their ability to protect against Alternaria early blight disease blight. The results show that the modified polymer with ZnONPs and beta-cyclodextrin (β-CD) and the modified polymer with ZnONPs (Chs-EF/ZnO NPs) provided the best protection, with DI values of 25 % and 12.5 %, respectively. Furthermore, it was discovered that the levels of carotenoids, chlorophyll a, and chlorophyll b in the infected plants had dropped by 52.6 %, 69.2 %, and 36.1 %, respectively. Chs-EF/ZnONPs were the most effective treatment, showing significant increases in the contents of chlorophyll a and b in infected plants by 120.8 % and 225.4 %, respectively. The study revealed that Chs-EF/ZnONPs exhibited a 131 % increase in the total phenolic content of plants, peroxidase (POD) activity (110.6 %), and a 347 % increase in polyphenol oxidase (PPO) activity, respectively, compared to healthy plants. Malondialdhyde (MDA) decreased by 50.7 %, 49.7 %, 43.4 %, 36.6 %, 31.7 %, and 7.5 % in response to Chs-EF/ZnONPs, Chs-EF/β-CD, Chs-EF, ZnONPs, Chitosan, and β-CD, respectively. Also, application of Chs-EF/ZnONPs, Chs-EF/β-CD, Chs-EF, ZnONPs, Chitosan, and β-CD reduced the production of H 2 O 2 by 77.5 %, 62.8 %, 62.5 %, 39.6 %, 22 %, and 15.1 %, respectively, compared to infected controls. We recommend considering these substances as promising candidates for agricultural use, as they may effectively serve as control agents against early blight caused by Alternaria solani., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Echinoderms possess unique biological traits that make them valuable models in immunology, regeneration, and developmental biology studies. As a class rich in active substances with significant nutritional and medicinal value, echinoderms face threats from marine pathogens, including bacteria, viruses, fungi, protozoa, and parasites, which have caused substantial economic losses in echinoderm aquaculture. Echinoderms counteract pathogen invasion through innate immunity and programmed cell death, in particular, with apoptosis being essential for eliminating infected or damaged cells and maintaining homeostasis in many echinoderm cell types. Despite the importance of this process, there is a lack of comprehensive and updated reviews on this topic. This review underscores that echinoderm apoptotic pathways exhibit a complexity comparable to that of vertebrates, featuring proteins with unique domains that may indicate the presence of novel signaling mechanisms. We synthesize current knowledge on how echinoderms utilize diverse transcriptional and post-transcriptional mechanisms to regulate apoptosis in response to pathogen infections and explore how pathogens have evolved strategies to manipulate echinoderm apoptosis, either by inhibiting it to create survival niches or by inducing excessive apoptosis to weaken the host. By elucidating the primary apoptotic pathways in echinoderms and the host-pathogen interactions that modulate these pathways, this review aims to reveal new mechanisms of apoptosis in animal immune defense and provide insights into the evolutionary arms race between hosts and pathogens., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microbial biofilms constitute a significant virulence factor and a substantial challenge in clinical environments due to their role in promoting antimicrobial resistance and their resilience to eradication efforts. Methicillin-resistant Staphylococcus aureus (MRSA) infections substantially increase healthcare costs, extend hospitalizations, and elevate morbidity and mortality rates. Therefore, developing innovative strategies to target and eliminate these bacteria and their biofilms effectively is imperative for robust epidemiological control. In this study, we evaluated the antibacterial and antibiofilm activities of cell-free supernatant (CFS) obtained from the Bacillus velezensis 1273 strain culture. Our data showed that CFS inhibited the growth of S. aureus ATCC 29213 and MRSA (clinical strain), with greater efficacy observed against S. aureus (1:16 dilution). Furthermore, CFS showed substantial potential in reducing biofilm formation in both strains (∼30 %) at subinhibitory concentrations. Additionally, the antibacterial activity against biofilm-formed cells showed that pure CFS treatment decreased the viability of S. aureus (60 %) and MRSA (45 %) sessile cells. We further demonstrated that CFS treatment induces the production of reactive oxygen species (ROS) and damages the membranes and cell walls of the pathogen cells. Genome analysis revealed the presence of genes encoding bacteriocins and secondary metabolites with antibacterial activity in the B. velezensis 1273 genome. These findings highlight the potential of probiotic bacterial metabolites as antibiofilm and anti-multidrug-resistant pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bacterial infections, particularly Salmonella , pose a significant health risk to neonates due to their underdeveloped immune systems. Understanding the immune responses in the neonatal intestine during S. Typhimurium infection is crucial for developing effective therapeutic and prevention strategies. This study found neonatal rats exhibited severe symptoms, including significant mortality, body weight loss, diarrhea, and bacterial load increases in the gastrointestinal tract and various organs, particularly in the ileum. Moreover, neonatal rats exhibited a high percentage of type 3 immune cells including Th17, γδT17, and ILC3 after S. Typhimurium infection. Furthermore, cintirorgon treatment during early life, the agonist of RORγt, significantly enhanced IL-17A-secreting type 3 immune response and alleviated the symptoms. Our data reveal targeting RORγt and IL-17A pathways may offer a promising therapeutic strategy for bacterial infections in neonatal populations.

Objectives: Reducing mortality from infectious diseases is an urgent global public health priority. Streptococcus pneumoniae, H. influenzae, and influenza virus are the three leading causes of lower respiratory infections (LRIs) death worldwide. Our objective was to assess the global burden of LRIs attributable to S. pneumoniae, H. influenzae, and influenza virus and explore the protective effectiveness of immunization programs., Methods: Data were retrieved from the Global Burden of Disease Study 2021 and World Health Organization United Nations Children's Fund Estimates of National Immunization Coverage. Locally weighted linear regression and Spearman correlation analysis were used to examine the associations between LRI mortality and vaccination coverage. Mixed-effects regression models were used to estimate the reduction in deaths that would be reduced by pneumococcal conjugate vaccine (PCV) and the H. influenzae type b (Hib) vaccine if all countries realized the Immunization Agenda 2030 (IA2030)., Results: In 2021, about 30.2% of the 2.18 million LRI-related deaths were attributed to three studied pathogens. From 1990 to 2021, the age-standardized mortality rate of LRIs was attributable to three pathogens decreased by more than half. In 2022, the global vaccination rates for PCV and Hib vaccines were 60.0% and 76.0%, respectively. The LRIs attributable to S. pneumoniae (r s = -0.45, P <0.001) and H. influenzae (r s = -0.47, P <0.001) decreased with the increasing vaccination coverage of PCV and Hib. By 2030, approximately 59.9% and 70.8% of countries worldwide will reach the IA2030 targets of 90% coverage for PCV and Hib, respectively. By that time, the number of deaths from LRIs in children attributable to S. pneumoniae and H. influenzae will decrease by 54.8% and 24.4%, respectively., Conclusions: Despite the declines in LRI mortality attributed to respiratory pathogens, substantial deaths still occurred in 2021. To advance toward achieving the IA2030 targets and further mitigate mortality associated with LRIs, intensified efforts by the international community and national health systems are imperative., Competing Interests: Declarations of competing interest The authors have no competing interests to declare., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The ever-evolving procedures of the practice of in vitro production (IVP) of embryos has well outpaced the ability of governments and institutions to create standardized rules and regulations around the global trade of these reproductive products. There are several challenges related to identifying and quantifying disease transmission risk in IVP embryos, not the least of which is a lack of published information. The International Embryo Technology Society (IETS) Health and Safety Advisory Committee (HASAC) has identified and addressed several of the challenges related to the potential for disease transmission via IVP embryos. This review will outline the current sanitation practices recommended by the IETS for in vivo derived (IVD) embryos, as well as the current practices at most in vitro fertilization (IVF) labs and will present the case that IVP embryos are not only just as safe as IVD embryos from a disease transmission risk standpoint, but perhaps may even be safer than IVD embryos, as the oocyte, zygote, and embryo in the in vitro production system has no contact with the downstream reproductive anatomy (fimbria, isthmus, oviduct, or uterus) of the donor female., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Soft tissue sarcomas (STS) often occur in the peri-pelvic region (proximal thigh, groin, gluteal region). A common complication following resection of STS is surgical site infection (SSI). The peri-pelvic site appears to be particularly problematic. Surgical site infections are associated with a high proportion of gram-negative and anaerobic micro-organisms. To date, there are no published recommendations for peri-operative antibiotic prophylaxis in pelvic STS resection. Therefore, the aim of this study was to determine the rate of SSI and the spectrum of micro-organisms detected in this region. Methods: In this monocentric study, 366 patients were retrospectively evaluated. All of these patients had undergone surgery for STS in the peri-pelvic and pelvic regions. Surgical site infections were recorded, and the microbial spectrum was analyzed. Results: There were 85 (23.2%) patients with SSI, and 188 revisions were required in these patients (2.21 per case). Swabs were sterile in 20% of clinically infected cases. In total, 36.5% of infections were polymicrobial. The most common bacteria were coagulase-negative staphylococci in 31.5%, followed by Enterococcus species in 13.3% and Escherichia coli in 7.7%. In total, 30.8% of the bacteria were gram-negative and 25.9% were anaerobic. Conclusions: Our results demonstrate the uniqueness of the bacterial spectrum of SSI after STS resection in the peri-pelvic region. In the authors' opinion, recommendations regarding the peri-operative antibiotic prophylaxis need to be adapted for the typical microbial spectrum at this site.

For the protection of human health and environment, there is a growing demand for high-performance, user-friendly biosensors for the prompt detection of pathogenic bacteria in samples containing various substances. We present a nanogap electrode-based purely electrical impedimetric sensor that utilizes the dielectrophoresis (DEP) mechanism. Our nanogap sensor can directly and sensitively detect pathogens present at concentrations as low as 1-10 cells/assay in buffers and drinking milk without the need for separation, purification, or specific ligand binding. This is achieved by minimizing the electrical double-layer effect and electrode polarization in nanogap impedance sensors, reducing signal loss. In addition, even at low DEP voltages, nanogap sensors can quickly establish strong DEP forces between the nanogap electrodes to control the spatial concentration of pathogens around the electrodes. This activates and stabilizes inter-electrode signal transmission along the nanogap-aligned pathogens, increasing sensitivity and reducing errors during repeated measurements. The DEP-enabled nanogap impedance sensor developed in this study is valuable for a variety of pathogen detection and monitoring systems including point-of-care testing (POCT) as it can detect pathogens in diverse samples containing multiple substances quickly and with high sensitivity, is compatible with complex solutions such as food and beverages, and provides highly reproducible results without the need for separate binding and separation processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In recent years, nanozymes have been widely used in the field of biosensing and food safety testing due to their advantages of low cost, high stability, easy modification and adjustable catalytic activity. However, how to reduce the signal interference generated by reducing substances, macromolecules and colored substances in the food matrix in nanozymes-based colorimetric sensing is still a major challenge. In this paper, using Listeria monocytogenes as a model analyte, sodium sulfonyl methacrylate (SBMA) polymers were modified onto cotton swabs by photothermal polymerization and combined with Listeria monocytogenes-specific aptamer (Apt1) to prepare swabs that can specifically capture and isolate Listeria monocytogenes from complex matrices (SBMA/Apt1 cotton swab). In addition, in combination with the inhibitory effect of the aptamer (Apt2) on the oxidase activity of Mn 3 O 4 NPs, a colorimetric biosensor based on nanozymes that can quantitatively, sensitively, and specifically identify Listeria monocytogenes in food products was constructed. The results showed that the colorimetric signal of the method was linear with the concentration of Listeria monocytogenes in the range of 2.83-2.83 × 10 5  CFU/mL, and the limit of detection was 2.64 CFU/mL, which can be used for the detection of Listeria monocytogenes in complex environments and food samples., Competing Interests: Declaration of competing interest The authors declare that they have no known conflict of interest or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objectives: To describe the symptoms, duration, severity, and microbiology of lower respiratory tract infection (LRTI) in outpatients., Methods: Prospective cohort study of adults in US primary or urgent care with a chief complaint of cough and symptoms consistent with LRTI. Baseline data included demographics, signs, symptoms, and PCR for 46 viruses and bacteria. The severity of symptoms reported for ≤28 days follow-up via diary and text message. The Bronchitis severity score assessed severity at baseline; overall severity was defined as the area under the symptom severity curve., Results: Of 718 patients with complete baseline data, 618 had valid PCR results, and 443 were followed until symptoms resolved. Of those with valid PCR, 100 (16.2%) had 1+ viruses detected, 211 (34.1%) had 1+ bacteria, and 168 (27.2%) had both. Symptoms more likely with viral or mixed infection included feverishness (36.7-38.4% vs. 18.5%), chills or sweats (36.0-38.1% vs. 17.9%), being generally unwell (78.2-81.3% vs. 64.9%), and myalgias (42.7-48.2% vs. 28.6%). Coloured sputum (42.9% vs. 23.2-29.5%) was more common with a bacterial infection. The mean duration of cough was 14.7 days with viruses (95% CI: 13.2-16.2), 17.3 with bacteria (95% CI: 15.9-18.6), 16.9 with mixed infection (95% CI: 15.2-18.6), and 18.4 with no detection (95% CI: 16.1-20.8). Overall severity of cough was lower for viral infections (20.9 points, 95% CI: 18.6-23.3) than for other groups (range 24.2-26.3). The most common potential bacterial pathogens were Haemophilus influenza (28.0%), Moraxella catarrhalis (16.2%), and Streptococcus pneumoniae (10.2%), whereas the most common viral pathogens were rhinovirus (17.3%), influenza (12.8%), SARS-CoV-2 (11.5%), and seasonal coronaviruses (8.1%)., Discussion: The mean duration of cough was 16.4 days. Consistent with European studies, the type of infection or potential pathogen was not an important predictor of the duration or severity of LRTI., (Copyright © 2024 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Nile tilapia ( Oreochromis niloticus ) and European sea bass ( Dicentrarchus labrax ) are economically significant species in Mediterranean countries, serving essential roles in the aquaculture industry due to high market demand and nutritional value. They experience substantial losses from bacterial pathogens Vibrio anguillarum and Streptococcus iniae , particularly at the onset of the summer season. The immune mechanisms involved in fish infections by V. anguillarum and S. iniae remain poorly understood. This study investigated their impact through experiments with control and V. anguillarum - and S. iniae -infected groups for each species. Blood samples were collected at 1, 3, and 7 days post bacterial injection to assess biochemical and immunological parameters, including enzyme activities (AST and ALT), oxidative markers (SOD, GPX, CAT, and MDA), and leukocyte counts. Further analyses included phagocyte activity, lysozyme activity, IgM levels, and complement C3 and C4 levels. Muscle tissues were sampled at 1, 3, and 7 days post injection to assess mRNA expression levels of 18 immune-relevant genes. The focus was on cytokines and immune-related genes, including pro-inflammatory cytokines ( TNF-α , TNF-β , IL-2 , IL-6 , IL-8 , IL-12 , and IFN-γ ), major histocompatibility complex components ( MHC-IIα and MHC-IIβ ), cytokine receptors ( CXCL-10 and CD4-L2 ), antimicrobial peptides ( Pleurocidin and β-defensin ), immune regulatory peptides ( Thymosin β12 , Leap 2 , and Lysozyme g ), and Galectins ( Galectin-8 and Galectin-9 ). β-actin was used as the housekeeping gene for normalization. Significant species-specific responses were observed in N. Tilapia and E. Sea Bass when infected with V. anguillarum and S. iniae , highlighting differences in biochemical, immune, and gene expression profiles. Notably, in N. Tilapia, AST levels significantly increased by day 7 during S. iniae infection, reaching 45.00 ± 3.00 ( p < 0.05), indicating late-stage acute stress or tissue damage. Conversely, E. Sea Bass exhibited a significant rise in ALT levels by day 7 in the S. iniae group, peaking at 33.5 ± 3.20 ( p < 0.05), suggesting liver distress or a systemic inflammatory response. On the immunological front, N. Tilapia showed significant increases in respiratory burst activity on day 1 for both pathogens, with values of 0.28 ± 0.03 for V. anguillarum and 0.25 ± 0.02 for S. iniae ( p < 0.05), indicating robust initial immune activation. Finally, the gene expression analysis revealed a pronounced peak of TNF-α in E. Sea Bass by day 7 post V. anguillarum infection with a fold change of 6.120, suggesting a strong species-specific pro-inflammatory response strategy. Understanding these responses provides critical insights for enhancing disease management and productivity in aquaculture operations.

Purpose: Neonatal bacterial meningitis (NBM) is a serious disease with high morbidity and mortality. This study aimed to establish a foundation for the selection of empirical antibiotics for NBM through an analysis of pathogen distribution and shift in antimicrobial resistant pattern., Patients and Methods: A retrospective cohort study on culture confirmed NBM from 2005 to 2022., Results: A total of 191 cases were enrolled, 48 for premature infants and 70 for early-onset meningitis. The incidence of NBM in first half and last half period was 0.2‰ and 0.24‰, while the mortality was 12.0% and 5.6% respectively. The top two pathogens were Escherichia coli ( E. coli ) (40.3%) and Group B Streptococcus (GBS) (29.8%). E. coli was the most common pathogen in both periods, Enterococcus (13.3% vs 0.9%, P < 0.05) and Staphylococcus aureus (8.4% vs 1.9%, P < 0.05) decreased, while GBS increased significantly in the periods [16.9% vs 39.8%, P < 0.001]. Gram-positive bacteria were more common in full term infants (53.8% vs 27.1%, P = 0.001), while gram-negative bacteria were more common in preterm infants (72.9% vs 46.2%, P = 0.001). All isolated GBS strains were susceptible to penicillin. On the other hand, less than 60% of E. coli were susceptible to third generation of cephalosporins, and were only susceptible to carbapenem or amikacin. This was mainly due to production of extended-spectrum beta-lactamase (ESBLs) which was higher in late-onset group than that in early-onset group (45.1% vs 19.2%, P = 0.026)., Conclusion: Incidence of NBM has not changed significantly over the last 2 decades. E. coli remains as the most common pathogen of NBM despite that GBS has increased in recent 9 years, especially in full-term infants. While all isolated GBS are susceptible to penicillin, over a third of E. coli strains are multidrug resistant due to production of ESBLs., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (© 2024 Zhu et al.)

The Chinese monal ( Lophophorus lhuysii ) is a large-sized and vulnerable (VU in IUCN) bird from southwestern China. This study applied 16S rRNA high-throughput sequencing to comprehensively examine the gut microbiota of captive Chinese monals (located in Baoxing, Sichuan, China) across varying seasons and life stages. Dominant bacterial phyla identified included Proteobacteria, Firmicutes , Bacteroidetes , and Actinobacteria . Significant seasonal and age-associated shifts were observed within specific bacterial groups, particularly marked by seasonal fluctuations in beta diversity. Moreover, linear discriminant analysis effect size (LEfSe) and functional predictions highlighted distinct winter signatures, indicating possible functional shifts in energy metabolism and disease resistance. In mid-aged adults, an expansion of Gamma-Proteobacteria suggested an elevated susceptibility of the gut microbiota of Chinese monals to chronic disorders and microbial imbalance. Putative pathogenic bacteria exhibited increased abundance in spring and summer, likely driven by temperature, host physiological cycles, interspecies interactions, and competition. These findings imply that the diversity, and structure of the gut microbiota in captive Chinese monals are strongly influenced by seasonal and age-related factors. The insights provided here are essential for improving breeding strategies and preventing gastrointestinal diseases in captivity.

Background: This study discusses the development trends and current applications of digital droplet PCR technology in pathogen diagnostics., Methods: A bibliometric analysis was conducted by retrieving droplet digital pcr related articles published between January 1,2012 to December31, 2023, from the Web of Science Core Collection., Results: A total of 3,513 authors from 66 countries published 508 research papers across 203 academic institutions. The keyword clustering network shows that the main advantages of ddPCR are absolute quantification, high sensitivity, and high tolerance to PCR inhibitors. The keyword timeline shows that current research hotspots include continuous pathogen nucleic acid monitoring, trace sample detection, and resistance mutation identification., Conclusion: The application of ddPCR in pathogen diagnosis is expanding, offering key advantages in absolute quantification, high sensitivity, and inhibitor tolerance, meeting emerging needs in nucleic acid monitoring and resistance detection., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interests, we do not have any possible conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Southern greater gliders (Petauroides volans) are endangered Australian marsupials for which there is little health data currently available. Chlamydia pecorum is the only reported pathogen of greater gliders and infects a broad range of hosts, including other marsupials, ruminants, swine and birds. Conjunctival and cloacal swabs collected from thirty-two southern greater gliders across southeastern New South Wales, Australia were screened for Chlamydia spp. by 23S real-time PCR. None of the ninety-six swabs tested returned a positive result for Chlamydia DNA. We found no evidence of Chlamydia infection among the four geographically disparate populations sampled, suggesting that Chlamydia may not pose a widespread health risk to southern greater gliders in this region, based on the current data. However, various clinical signs of ophthalmic pathology were observed, warranting further investigation to determine their underlying aetiology., Competing Interests: Declarations. Ethics approval: All procedures were approved by the University of Wollongong research ethics committee (approval number AE19/02) and performed in accordance with the ethical standards laid out in the NSW Animal Research Act and the Australian Code for the Care and Use of Animals for Scientific Purposes. This research was conducted under a NSW Scientific Licence (SL101968) granted by the Department of Climate Change, Energy, the Environment and Water. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Seed-based restoration methods are increasingly recognized as a relevant tool contributing to halt and reverse the loss of seagrass meadows while providing genetic and evolutionary benefit for the conservation of these habitats. Ad-hoc protocols aimed at maximizing the survival of plantlets obtained from seeds in cultivation systems are therefore required. Previous trials of seedling culture of Posidonia oceanica , the dominant seagrass of the Mediterranean Sea, recorded up to 40% loss due to mould development. In this study we aim to (i) identify the putative causal agents of seed decay and (ii) test the efficacy of copper sulphate (0.2 and 2 ppm) and of tannin-based products derived from chestnut, tara and quebracho in reducing seed and seedling decay, while assessing possible phytotoxic effects on plant development. Halophytophthora lusitanica, H. thermoambigua and a putative new Halophytophtora species were identified as possible causal agents of seed loss. The antimicrobial agents (copper and tannins) reduced seed contamination by 20%, although copper sulphate at 2 ppm strongly inhibited the root growth. Among tannins, chestnut and tara reduced seeds germination by up to 75% and decreased shoot and root development, while quebracho showed a less severe phytotoxic effect. The use of copper sulphate at 0.2 ppm is therefore recommended to prevent P. oceanica seedling loss in culture facilities since it reduces seed contamination with no phytotoxic effects. Our results contribute to improving the seedling culture of one the key species of the Mediterranean Sea, increasing propagule availability for restoration purposes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Alagna, Giacalone, Zenone, Martinez, D’Anna, Buffa, Cavalca, Poli, Varese, Prigione and Badalamenti.)

Detection of microorganisms from bovine using real-time PCR (Dembo PCR) is a comprehensive detection technique that was developed to detect pathogens causing bovine diseases. In Japan, the definitive tests for monitored infectious diseases, which are defined by law, are carried out at government agencies. On the other hand, the existence of pathogens other than monitored infectious diseases are not well understood. From the perspective of livestock quarantine, it is important to make it possible to identify pathogens other than monitored infectious diseases, so we extended the existing Dembo PCR system in this study. In particular, the number of targets other than monitored infectious disease was increased. The new version of Dembo PCR may be useful in elucidating new pathologies associated with multiple pathogens.

Induced systemic resistance (ISR) is a crucial concept in modern agriculture, explaining plant defense mechanisms primed by rhizosphere stimuli and activated by subsequent infections. Biological factors contributing to ISR generally include plant growth-promoting microbes3 (PGPM). Bacillus spp., Pseudomonas spp., and Trichoderma spp. have been extensively studied for their plant growth-promoting characteristics and ISR effect against above-ground pathogens and insect infestations. These phenomena elucidate the bottom-up effects of how beneficial rhizosphere microbes help plants resist above-ground attacks. Conversely, soil microbiome analysis in the rhizosphere of plants infected by above-ground pathogens has shown increased beneficial microbes in the soil, a phenomenon termed 'soil legacy effects'. This represents the top-down effects of above-ground attackers on plants' rhizosphere environments. Interestingly, recent studies have shown that above-ground stimuli not only recruit PGPM in the rhizosphere but also that these PGPM influence plant defense responses against subsequent pathogen infections. This can be seen as a four-step plant defense mechanism involving above-ground attackers, host plants, rhizosphere microbes, and subsequent attacks. This represents an active defense mechanism that overcomes the limitations of sessile plants. This review summarizes plant ISR mechanisms in terms of triple inter-organism interactions and provides molecular evidence for each step., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Jung, Ahn, Kim and Riu.)

Strophariarugosoannulata is an important edible mushroom in China, but green mold disease has caused significant production and economic losses. In this study, two new pathogens Trichodermastrophariensis and T.viridistromatis were identified as the causal agents of this disease. During October-November 2023, six strains of the fungal pathogen were isolated from infected fruiting bodies of S.rugosoannulata and identified based on morphological characteristics and molecular phylogenetic analyses of internal transcribed spacer (nrITS), the second largest RNA polymerase II subunit ( rpb2 ) and the partial translation elongation factor 1-alpha ( tef1-α ) region. The representative isolates of the pathogenic green mold Trichoderma species were used to perform a pathogenicity test with spore suspensions, resulting in symptoms similar to those observed in the cultivated field. The same pathogens were successfully re-isolated, thereby fulfilling Koch's postulates. Detailed morphological descriptions, illustrations, culture characteristics, and comparisons with morphologically similar and closely related species are provided., Competing Interests: The authors have declared that no competing interests exist., (Entaj Tarafder, Zhang Wenjun, Samantha C. Karunarathna, Abdallah M. Elgorban, Man Huilian, Wu Nan, Xiangyu Zeng, Wang Yong, Feng-Hua Tian.)

Reports on the application of metagenomic next-generation sequencing (mNGS) in adult patients with severe pneumonia after cardiac surgery remain limited. This study aimed to evaluate the clinical outcomes of mNGS analysis of bronchoalveolar lavage fluid (BALF) in such patients.A retrospective cohort study was conducted on adult patients with severe pneumonia after cardiac surgery. Samples were collected from patients in the surgical intensive care unit (SICU) of Sichuan Provincial People's Hospital between January 2019 and March 2024. Upon diagnosis of severe pneumonia, bronchoalveolar lavage fluid was obtained via bronchoscopy within 24 h. The mNGS group was composed of patients tested using mNGS and conventional microbiological tests. BALF was detected only by the conventional microbiological test (CMT) method in the CMT group, which involved examining bacterial and fungal smears and cultures at least. We reviewed a total of 4,064 cardiac surgeries, and based on the inclusion criteria, a total of 113 adult patients with severe pneumonia after cardiac surgery were included in this study. The overall positive rate detected by mNGS was significantly higher than that of the culture method (98% vs. 58%, P<0.0001). After receipt of the microbiological results, the mNGS group exhibited a higher incidence of antibiotic adjustments in comparison to the CMT group (P = 0.0021). After adjusting the treatment plan based on microbial testing results, the mNGS group showed an improvement in ventilator-free days within 28 days (P = 0.0475), with a shorter duration of invasive ventilation compared to the CMT group (P = 0.0208). The detection of mNGS can significantly improve the Acute Physiology and Chronic Health Evaluation-II (APACHE II) score (P = 0.0161) and Sequential Organ Failure Assessment (SOFA) score (P = 0.0076) on the 7th day after admission to the SICU. In this study, the mNGS group showed signs of having a positive impact on the length of stay in ICU (median: 9 days, IQR: 7-10 days vs. median: 10 days, IQR: 8-13.75 days, P = 0.0538), length of stay in Hospital (median: 20 days, IQR: 17-28 days vs. median: 25 days, IQR: 18-29 days, P = 0.1558), mortality in 28 days (19% vs. 20%, P = 0.8794), in-hospital mortality (19% vs. 22%, P = 0.7123); however, statistical analysis did not confirm these differences to be significant. mNGS could serve as a valuable complement to conventional diagnostic approaches in adult patients with severe pneumonia after cardiac surgery, potentially improving diagnostic accuracy and leading to more precise and timely interventions, with significant potential to inform clinical decision-making and enhance patient outcomes., Competing Interests: Declarations. Ethics approval and consent to participate: The protocol used in this retrospective study was reviewed and approved by the Ethical Review Committee of Sichuan Provincial People’s Hospital (No: 2024-30). The study was carried out in accordance with the Declaration of Helsinki and all data were anonymous before analysis. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Pest cockroaches share urban habitats with us; their prevalence in urban areas prompts concerns regarding their effect on human health, as synanthropic cockroaches often host pathogenic microorganisms. Nonetheless, microbial associates in these insects can also be related to their biology, contributing to their physiological homeostasis and reproductive success. In this article, we present in detail, for the first time, the bacterial community associated with the oriental cockroach Blatta orientalis, one of the world's five most prominent pest cockroaches. We report the composition of the communities of bacteria found over the exoskeleton and inside the gut of this global pest. We collected B. orientalis in Santiago, Chile's capital city, and the urban nucleus in this country. We conducted DNA extractions and metabarcoding analysis. We found diverse bacterial lineages, including mutualist symbiotic strains, and microorganisms considered pathogenic to humans. We also analyzed the metabolic functions of the bacterial communities identified and discussed the role of B. orientalis as a reservoir and vector of pathogens in urban areas. We discuss to what extent the diversity of functions of the microbial community associated with cockroaches may contribute to emergent properties enabling these insects to inhabit human-modified habitats.

Background: There may be potential associations between various pathogens, antibody immune responses, and breast cancer (BC), but the specific mechanisms and causal relationships remain unclear., Methods: First, multiple Mendelian randomization (MR) methods were used for univariable MR analysis to explore potential causal relationships between 34 antibody immune responses (related to 12 pathogens), 46 antibody immune responses (related to 13 pathogens), antibody responses post-COVID-19 vaccination, 731 immune cell types, and various BC subtypes (including overall BC, ER-positive, ER-negative, Luminal A, Luminal B, Luminal B HER2-negative, HER2-positive, and triple-negative BC). The primary results were then subjected to reverse MR analysis, heterogeneity testing using Cochran's Q, and horizontal pleiotropy testing. Robust findings were further used to design mediation pathways involving antibody immune responses, immune cells, and BC. After adjusting the effect estimates using multivariable MR (MVMR), a two-step mediation analysis was conducted to explore mediation pathways and mediation proportions. Finally, linkage disequilibrium score regression (LDSC) was applied to analyze the genetic correlation between phenotypes along mediation pathways, and cross-phenotype association analysis (CPASSOC) was performed to identify pleiotropic SNPs among three phenotypes along these pathways. Bayesian colocalization tests were conducted on pleiotropic SNPs using the multiple-trait-coloc (moloc)., Results: We identified potential causal relationships between 15 antibody immune responses to 8 pathogens (Hepatitis B virus, Herpes Simplex Virus 2, Human Herpesvirus 6, Polyomavirus 2, BK polyomavirus, Cytomegalovirus, Helicobacter pylori, Chlamydia trachomatis), 250 immune cell phenotypes, and various BC subtypes. MVMR-adjusted mediation analysis revealed four potential mediation pathways. LDSC results showed no significant genetic correlation between phenotypes pairwise. CPASSOC analysis identified two potential mediation pathways with common pleiotropic SNPs (rs12121677, rs281378, rs2894250). However, none of these SNPs passed the Bayesian colocalization test by moloc. These results excluded horizontal pleiotropy, stabilizing MR analysis results., Conclusion: This study utilized MR methods to analyze potential causal relationships between various antibody immune responses, immune cell types, and BC subtypes, identifying four potential regulatory mediation pathways. The findings of this study offer potential targets and research directions for virus-related and immunotherapy-related studies, providing a certain level of theoretical support. However, limitations such as GWAS sample size constraints and unclear specific pathophysiological mechanisms need further improvement and validation in future studies., Competing Interests: Declarations Ethics approval and consent to participate Since this study utilized de-identified public data, there was no need for additional approval. All original ethical approvals are available in the original literature and on the website. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Since the outbreak of the new coronavirus, point-of-care diagnostics based on nucleic acid testing have become a requirement for the development of pathogen diagnostics, which require the ability to accurately, rapidly, and conveniently detect pathogens. Conventional nucleic acid amplification techniques no longer meet the requirements for pathogen detection in low-resource, low-skill environments because they require specialist equipment, complex operations, and long detection times. Therefore, recombinant polymerase amplification (RPA) is becoming an increasingly important method in today's nucleic acid detection technology because it can amplify nucleic acids in 20-30 minutes at a constant temperature, greatly reducing the dependence on specialist equipment and technicians. RPA products are primarily detected through methods such as real-time fluorescence, gel electrophoresis, lateral flow assays (LFAs), and other techniques. Among these, LFAs allow for the rapid detection of amplification products within minutes through the visualization of results, offering convenient operation and low cost. Therefore, the combination of RPA with LFA technology has significant advantages and holds broad application prospects in point-of-care (POC) diagnostics, particularly in low-resource settings. Here, we focus on the principles of RPA combined with LFAs, their application to pathogen diagnosis, their main advantages and limitations, and some improvements in the methods., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhao, Wei, Ye, Cao, Zhou, Xie, Qing and Chen.)

Objective: To investigate the incidence, pathogen distribution, and antibiotic susceptibility of early- and late-onset neonatal bacteremia, and to analyze pathogen trends before and during the COVID-19 pandemic., Methods: Between January 2016 to December 2022, we collected 879 blood and cerebrospinal fluid specimens from newborns with bacteremia. Bacterial identification used biochemical methods and MALDI-TOF, and antibiotic susceptibility was tested with the VITEK 2 system. Incidence per 1,000 admissions was calculated with Wilson's 95% confidence intervals, and categorical variables were compared using χ²-test or Fisher's exact test., Results: Early-onset bacteremia incidence was 2.6 per 1,000 admissions, and late-onset bacteremia was 26.3, with a significant decline from 70.7 to 10.5 per 1,000 admissions over the study period. Late-onset bacteremia was more common before COVID-19, while early-onset bacteremia increased during the pandemic. The top five pathogens were CoNS(39.9%), E. faecalis(17.7%), E. faecium(13.7%), E. coli(8.4%), and GBS(5.8%). During the COVID-19 pandemic, the incidence of CoNS and S. aureus infections significantly decreased. Throughout the entire study period, CoNS and S. aureus showed high resistance to penicillin G and erythromycin but were sensitive to vancomycin and linezolid. E. faecalis and E. faecium were susceptible to vancomycin, linezolid, and teicoplanin but resistant to erythromycin, tetracycline, and rifampin. MRCoNS and MRSA were detected in 72.7% and 31.0% of isolates, respectively. Resistance rates of E. faecium and E. faecalis to ampicillin decreased significantly, clindamycin resistance in GBS decreased during the pandemic., Conclusion: This study highlights notable shifts in neonatal bacteraemia patterns during the COVID-19 Pandemic that were likely influenced by increased infection control and disruptions in maternal care, leading to changes in pathogen distribution and antimicrobial resistance., Competing Interests: Declarations Ethical approval We confirm that our study strictly followed all ethical standards set forth in the Declaration of Helsinki. The study protocol was approved by the Ethics Review Committee of Wuhan Children’s Hospital (NO.2024R050-E01). Given the retrospective design of this study, it did not interfere with standard medical care or infringe upon patient rights, nor did it pose any additional risk to participants. We ensured the protection of patient identities through coding, with medical records securely stored and accessible only to authorized researchers.The Ethics Review Committee of Wuhan Children’s Hospital has granted a waiver of informed consent. Consent for publication All authors declared no conflict of interest existed in this work. All authors are aware of and agree to the content of the paper and their being listed as a co-author of the paper. Clinical trial registration Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In this study, we analyzed bioaerosol emission characteristics and potential risks of antimicrobial resistance (AMR) during composting using the impaction culture method and metagenomic sequencing. The results showed that the highly saturated water vapor in the emission gas mitigated particulate matter emission during the thermophilic period. About the bioaerosols, the airborne aerobic bacterial emissions were suppressed as composting enters the mature period, and the airborne fungi are usually present as single-cell or small-cell aggregates (< 3.3 µm). In addition, the microbial community structure in bioaerosols was stable and independent of composting time. Most importantly, the PM 2.5 in bioaerosols contained large amounts of antibiotic resistance genes (ARGs), potential pathogens, and multidrug resistant pathogens, which were diverse and present in high concentrations. Among them, ARGs concentrations encoding 21 antibiotics ranged from - 4.50 to 0.70 ppm/m 3 (Log 10 ARGs). Among the 89 potential human pathogens detected, Escherichia coli, Salmonella enterica, Klebsiella pneumoniae, and Staphylococcus aureus were the only culturable potentially multidrug resistant pathogens carrying multiple ARGs encoding resistance at high concentrations (- 0.57 to 1.15 ppm/m 3 (Log 10 ARGs)), and were more likely to persist and multiply in oligotrophic environments. Our findings indicate that composting technology can transfer AMR from solid compost to gas phase and increase the risk of AMR transmission., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)