Bacterial Killing Ability Research Articles

An ACE inhibitor reduces bactericidal activity of human neutrophils in vitro and impairs mouse neutrophil activity in vivo.

Angiotensin-converting enzyme inhibitors (ACEIs) are used by millions of patients to treat hypertension, diabetic kidney disease, and heart failure. However, these patients are often at increased risk of infection. To evaluate the impact of ACEIs on immune responses to infection, we compared the effect of an ACEI versus an angiotensin receptor blocker (ARB) on neutrophil antibacterial activity. ACEI exposure reduced the ability of murine neutrophils to kill methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Klebsiella pneumoniae in vitro. In vivo, ACEI-treated mice infected with MRSA had increased bacteremia and tissue bacteria counts compared to mice treated with an ARB or with no drug. Similarly, ACEIs, but not ARBs, increased the incidence of MRSA-induced infective endocarditis in mice with aortic valve injury. Neutrophils from ACE knockout (KO) mice or mice treated with an ACEI produced less leukotriene B4 (LTB4) upon stimulation with MRSA or lipopolysaccharide, whereas neutrophils overexpressing ACE produced more LTB4 compared to wild-type neutrophils. As a result of reduced LTB4 production, ACE KO neutrophils showed decreased survival signaling and increased apoptosis. In contrast, neutrophils overexpressing ACE had an enhanced survival phenotype. Last, in a cohort of human volunteers receiving the ACEI ramipril for 1 week, ACEI administration reduced neutrophil superoxide and reactive oxygen species production and neutrophils isolated from volunteers during ramipril treatment had reduced bactericidal activity. Together, these data demonstrate that ACEI treatment, but not ARB treatment, can reduce the bacterial killing ability of neutrophils.

Neddylation Alleviates Methicillin-Resistant Staphylococcus aureus Infection by Inducing Macrophage Reactive Oxygen Species Production.

Neddylation, a posttranslational modification in which NEDD8 is covalently attached to target proteins, has emerged as an endogenous regulator of innate immunity. However, the role of neddylation in methicillin-resistant Staphylococcus aureus (MRSA) infection remains unknown. In this study, we found that neddylation was activated after MRSA infection in vivo and in vitro. Inhibition of neddylation with MLN4924 promoted injury of liver and kidneys in C57BL/6 mice with MRSA bloodstream infection and increased mortality. Blockade of neddylation, either pharmacologically (MLN4924, DI591) or through the use of Uba3 small interfering RNA, inhibited Cullin3 neddylation and promoted Nrf2 accumulation, thus reducing reactive oxygen species (ROS) induction and bacterial killing ability in mouse peritoneal macrophages. In summary, our findings suggest that activation of neddylation in macrophages plays a critical protective role against MRSA infection by increasing ROS production, partially by signaling through the NEDD8-Cullin3-Nrf2-ROS axis. Furthermore, our results may provide a new non-antibiotic treatment strategy for MRSA infection through targeting of neddylation.

Who Rules Over Immunology? Seasonal Variation in Body Temperature, Steroid Hormones, and Immune Variables in a Tegu Lizard.

Multiple factors can influence the immune response of ectothermic vertebrates, including body temperature (Tb), gonadal steroids, and seasonality, in ways that are thought to reflect trade-offs between energetic investment in immunity versus reproduction. Hibernating tegu lizards (Salvator merianae) are a unique model to investigate how immunocompetence might be influenced by different factors during their annual cycle. We assessed immunological measures (plasma bacterial killing ability, total and differential leukocyte count), plasma hormone levels (testosterone in males, estradiol and progesterone in females, and corticosterone [CORT] in both sexes), Tb, and body condition from adult tegus during each stage of their annual cycle: reproduction, post-reproduction/preparation for hibernation, and hibernation. Our hypothesis that immune traits present higher values during the reproductive phase, and a sharp decrease during hibernation, was partially supported. Immune variables did not change between life history stages, except for total number of leukocytes, which was higher at the beginning of the reproductive season (September) in both males and females. Average Tb of the week prior to sampling was positively correlated with number of eosinophils, basophils, monocytes, and azurophils, corroborating other studies showing that when animals maintain a high Tb, there is an increase in immune activity. Surprisingly, no clear relationship between immune traits and gonadal steroids or CORT levels was observed, even when including life history stage in the model. When gonadal hormones peaked in males and females, heterophil: lymphocyte ratio (which often elevates during physiological stress) also increased. Additionally, we did not observe any trade-off between reproduction and immunity traits, sex differences in immune traits, or a correlation between body condition and immune response. Our results suggest that variation in patterns of immune response and correlations with body condition and hormone secretion across the year can depend upon the specific hormone and immune trait, and that experienced Tb is an important variable determining immune response in ectotherms.

LPS-induced immunomodulation and hormonal variation over time in toads.

The inflammatory response is a complex process that relies on interactions among multiple endocrine and immune modulators. Studies incorporating time-related and integrative endocrine and immune responses to an immune challenge might shed light on the characterization of the phases of the inflammatory response in anurans. The present study investigated time-related changes (1, 3, 6, and 18 h post-challenge) in plasma corticosterone (CORT), melatonin (MEL) and testosterone (T) levels, phagocytosis percentage (PP), plasma bacterial killing ability (BKA), and neutrophil to lymphocyte ratio (NLR) following a lipopolysaccharide (LPS) immune challenge in Rhinella diptycha toads. Our results showed the response to LPS injection was characterized by increased CORT, PP, BKA, and NLR, with a concomitant decrease in plasma MEL and T. Increased CORT was more pronounced at 6and 18 h, while increased NLR was observed only 18 h post-LPS injection. Meanwhile, plasma MEL and T decreased independently of the time post-LPS injection. Additionally, toads in better body condition showed higher BKA and PP in the LPS-treated group, regardless of the time postinjection. Our results show that toads (R. diptycha) were sensitive to the LPS challenge, mounting an inflammatory response, which started quickly (after 1 h) and developed over time and was influenced by body condition. These results demonstrate a time-related hormonal and immune variation as a consistent pattern of activation of the immune system, as well as of hypothalamic-pituitary-adrenal/interrenal and immune-pineal axes following an immune challenge more deeply studied in mammals, suggesting the evolutionary conservation of the regulatory mechanisms for tetrapod vertebrates.

Reference Intervals in Combined Veterinary Clinical Examinations of Male Black-Spotted Pond Frogs (Pelophylax nigromaculatus).

Simple SummaryThe reference intervals (RIs) of immunity, serum components, bone mineral density (BMD), and body composition in 151 males of Pelophylax nigromaculatus were established. These analyses are easily replicable and can safely and accurately diagnose the physiological condition of animals. The use of combined examination allows for the establishment of a successful conservation strategy through the identification of conservation problems in many vertebrate groups.In conservation physiology, analyzing the physiological response of an organism to understand its ability to adapt to environmental changes is a key technique in establishing a successful conservation strategy. Veterinary clinical examinations determine the physiological condition of animals accurately and safely, and this examination is synergistic when combined. The accuracy and safety of a clinical examination makes it advantageous for use in amphibians with high species diversity and numerous endangered species. However, it is necessary to establish a reference interval (RI) for precise interpretations and identification of animals with abnormalities through individual unit testing. We have established RIs for the immunity, serum components, bone mineral density (BMD), and body composition of black-spotted pond frogs (Pelophylax nigromaculatus). Black-spotted pond frogs are a common species and are widely distributed in East Asia, with suitable characteristics for environmental monitoring. Serum was extracted from 151 male frogs to establish the RI for bacterial killing ability in order to represent immunity. We also used the serum to establish an RI of ten additional serum components to determine the nutritional status, organ function status, body osmotic pressure, and homeostasis conditions. The BMD and three body composition measures for diagnosing food intake status and nutritional condition were measured using dual energy X-ray absorptiometry. The RI was recorded as the mean ± standard deviation, median, first (25%) to third (75%) quantile range, 95% confidence interval of the mean and median, and the 95% percentile (2.5%–97.5% range) of all components. The use of combined clinical veterinary examinations aids our understanding of the physiological conditions of an individual according to biotic and abiotic factors on a complex spatiotemporal scale in an ecosystem.

Bacteria Strain‐Specific Chlorinated Lipid Production in Neutrophils

Neutrophils play vital roles in host defense against infections and acute inflammation. They are the major type of white blood cells first to arrive at the site of infection. Neutrophils are equipped with three major microorganism-killing mechanisms: phagocytosis, release of antibacterial enzymes, and formation of neutrophil extracellular traps (NET). During neutrophil activation, the primary granules release myeloperoxidase (MPO), which is a major antibacterial enzyme. MPO catalyzes hydrogen peroxide and chloride to produce hypochlorous acid (HOCl). HOCl has a significant role as an antimicrobial agent and can also have deleterious effects on host cells. Previous studies have shown that HOCl targets vinyl ether bond at the sn-1 position of plasmalogen phospholipids to liberate 2-chlorofatty aldehyde (2-ClFALD). 2-ClFALD further gets oxidized to 2-chlorofatty acids (2-ClFA). 2-ClFALD and 2-ClFA are linked with inflammatory diseases such as endotoxemia, sepsis and atherosclerosis. The roles of 2-ClFALD and 2-ClFA have been explored in endothelial dysfunction, endoplasmic reticulum stress, apoptosis in monocytes, neutrophil chemotaxis and as a mediator of NET formation. Here we tested the hypothesis that the neutrophils are activated by bacterial stimuli to produce 2-ClFALD and 2-ClFA and the production may vary with the bacterial strain. We co-cultured human neutrophils with CFT073 and JM109, pathogenic and non-pathogenic E.coli strains, respectively. 2-ClFALD production was significantly increased when neutrophils were incubated with either E.coli strains compared to control neutrophils. Interestingly, 2-ClFA was only increased in CFT073 co-culture. We have also observed that neutrophils killed more JM109 cells compared to CFT073 cells. However, CFT073 induced more NET formation suggesting the higher 2-ClFA might be mediating NET formation. Furthermore, JM109 was more susceptible to killing by 2-ClFALD and 2-ClFA indicating their potential role in bactericidal activity. 2-ClFA and 2-ClFALD production was significantly reduced in neutrophil and E.coli co-cultures by the addition of amino triazole (ATZ), an MPO inhibitor, and glutathione (GSH) that makes GSH-FALD adduct as a 2-ClFALD scavenger. ATZ treatment also reduced bacterial killing ability suggesting that MPO is an essential component of neutrophil antibacterial mechanisms. In contrast, GSH supplementation increased bacterial killing capacity. Taken together, these findings suggest that specific bacterial strains play a significant role in neutrophil derived chlorinated lipid production, which may also differentiate their susceptibility to neutrophil killing.

Influence of Ni2+ ions on the structural, morphological, photoluminescence, photo-catalytic and anti-bacterial studies of Cd0.9Zn0.1S nanostructures.

The present investigation reported the controlled synthesis of Cd0.9Zn0.1S and Cd0.89Zn0.1Ni0.01S nanostructures by simple chemical co-precipitation route. The XRD analysis confirmed the cubic structure of CdS on Zn doped CdS and Zn, Ni dual doped CdS without any secondary/impurity phases and no alteration in CdS cubic phase was noticed by Zn/Ni addition. The shrinkage of crystallite size from 69 to 43 Å and the variation in lattice constants and micro-strain were described by the addition of Ni and the defects associated with Ni2+ ions. Microstructural and optical studies of the prepared films were carried out using scanning electron microscope (SEM), UV-visible spectrometer and photoluminescence (PL) spectra. The enhanced optical absorbance in the visible wavelength and the reduced energy gap by Ni substitution showed that Cd0.89Zn0.1Ni0.01S nanostructures are useful to improve the efficiency of opto-electronic devices. The functional groups of Cd-S/Zn-Cd-S/Zn/Ni-Cd-S and their chemical bonding were verified by Fourier transform infrared (FTIR) studies. The elevated visible PL emissions such as blue and green emissions by Ni addition was explained by decreasing of crystallite size and generation of more defects. Zn, Ni dual doped CdS nanostructures are identified as the probable an efficient photo-catalyst for the degradation of methylene blue dye. The liberation of more charge carriers, better visible absorbance, improved surface to volume ratio and the creation of more defects are accountable for the current photo-catalytic activity in Zn/Ni doped CdS which exhibited better photo-catalytic stability after sex cycling process. The better bacterial killing ability is noticed in Ni doped Cd0.9Zn0.1S nanostructure which is due to the collective effect of lower particle/grain size and also higher ROS producing capacity.

Stress Response, Immunity, and Organ Mass in Toads (Rhinella diptycha) Living in Metal-Contaminated Areas.

Mining is one of the main activities that drive the economy of Brazil. Mining activity is associated with risk of contamination of environment and local fauna by metals. Amphibians have a life cycle that requires a transition between aquatic and terrestrial environments, increasing their vulnerability to metal contamination in the water and substrate. Metals are ubiquitous, with high bioaccumulative and biomagnifying potential, and may lead to immune and endocrine disruption. In this study, we analyzed two different components of the innate immune response, bacterial killing ability (BKA) and phytohemagglutinin edema (PHA), and two stress biomarkers, corticosterone plasma levels (CORT) and the neutrophil to lymphocyte ratio (N:L), of toads (Rhinella diptycha) living in places contaminated by metals. Blood samples were collected pre- and post-restraint (1h), followed by an immune challenge with PHA and tissue collection (liver, spleen, and kidneys). Toads liver metal bioaccumulation did not correlate with the immune response or stress biomarkers. Post-restraint, animals had increased CORT and reduced BKA, independently of the collection site, and these variables were not correlated with liver metal bioaccumulation. Interestingly, toads with the larger spleen (immune organ) showed increased N:L post-restraint and greater edema after the PHA challenge. Our results indicate that toads living in metal-contaminated environments responded to acute stressor, activating the hypothalamic-pituitary-interrenal axis and the immune response. Keep tracking the physiological variables of these animals and the presence of metals in the environment and tissues should provide valuable health status indicators for the population, which is vital for proposing amphibian conservation strategies in these areas.

Short-term stressors and corticosterone effects on immunity in male toads (Rhinella icterica): A neuroimmune-endocrine approach

In the last decades, it is growing the idea that stress-induced immunomodulation is bimodal: with acute stress associated with enhancing effects while chronic stress with suppressive effects. However, the immune-endocrine interactions and its implications are often overlooked in ectotherms. We investigated the impact of corticosterone (CORT) treatment and short-term stressors on CORT plasma levels and the immunity of male toads (Rhinella icterica), using three distinct protocols: restraint, immune challenge (with lipopolysaccharide, LPS), and CORT transdermal application (TA). Our results showed increased CORT and neutrophil: lymphocyte ratio (NLR) regardless of the stress input (restraint, LPS challenge) or CORT TA. In the meantime, the bacterial killing ability (BKA) was not affected by any treatment, suggesting this immune parameter might be a more constitutive and robust response. Interestingly, the cellular immune response showed distinct patterns. Increased phagocytosis of blood leukocytes and phytohemagglutinin edema followed LPS and CORT TA (15 ​μg), respectively. In contrast, the phagocytosis of peritoneal leukocytes decreased after CORT TA (1 and 10 ​μg), indicating that short-term increases in CORT levels might impair local immune function. Such differences in cellular immunity might also be associated with CORT doses or the interaction between CORT and other immune mediators, such as melatonin, testosterone, and cytokines. Overall, our results highlight the immune-enhancing effects of the acute stress response and CORT TA, and the complexity of the immune-endocrine interaction in anurans. It also highlights the relevance of investigating distinct contexts for CORT increase arising from different situations, as well as diverse immune components for a better understanding of the stress-induced immunomodulation.

Solvent-Free Fabrication of Self-Regenerating Antibacterial Surfaces Resisting Biofilm Formation.

Biofilm formation on indwelling medical devices is a major cause of hospital-acquired infections. Monofunctional antibacterial surfaces have been developed to resist the formation of biofilms by killing bacteria on contact, but the adsorption of killed bacterial cells and debris gradually undermines the function of these surfaces. Here, we report a facile approach to produce an antibacterial surface that can regenerate its function after contamination. The self-regenerating surface was achieved by sequential deposition of alternating antibacterial and biodegradable layers of coating using a solvent-free initiated chemical vapor deposition method. As the top antibacterial layer gradually loses its killing ability due to the accumulation of debris, the underlying biodegradable layer degrades, shedding off the top surface layers and exposing another fresh antibacterial surface. Urinary catheters coated with monofunctional and self-regenerating antibacterial coatings both showed more than 99% bacterial killing ability at the initial antibacterial test, but the monofunctional surface lost its killing ability after continued exposure to concentrated bacterial solution, whereas the self-regenerating surfaces regained strong bacterial killing ability after prolonged exposure. Employing poly(methacrylic anhydride) and its copolymers with varied composition as the degrading layer, the degradation kinetics can be well-tailored and the self-regeneration duration spanned from minutes to days. The designed self-regenerating antibacterial surfaces could provide an effective approach to resist biofilm formation and extend the service life of indwelling medical devices.

Understanding metrics of stress in the context of invasion history: the case of the brown treesnake (Boiga irregularis).

Invasive species can exert rapid depletion of resources after introduction and, in turn, affect their own population density. Additionally, management actions can have direct and indirect effects on demography. Physiological variables can predict demographic change but are often restricted to snapshots-in-time and delayed confirmation of changes in population density reduces their utility. To evaluate the relationships between physiology and demography, we assessed metrics of individual and demographic stress (baseline and 1-h corticosterone (CORT), body condition and bacterial killing ability) in the invasive snake Boiga irregularis on Guam collected in intervals of 10-15years. We also assessed potential discrepancies between different methods of measuring hormones [radioimmunoassay (RIA) versus enzyme immunoassay (EIA)]. The magnitude of difference between RIA and EIA was negligible and did not change gross interpretation of our results. We found that body condition was higher in recent samples (2003 and 2018) versus older (1992-93) samples. We found corresponding differences in baseline CORT, with higher baseline CORT in older, poorer body condition samples. Hormonal response to acute stress was higher in 2018 relative to 2003. We also found a weak relationship between circulating CORT and bacterial killing ability among 2018 samples, but the biological significance of the relationship is not clear. In an effort to develop hypotheses for future investigation of the links between physiology and demography in this and other systems, we discuss how the changes in CORT and body condition may reflect changes in population dynamics, resource availability or management pressure. Ultimately, we advocate for the synchronization of physiology and management studies to advance the field of applied conservation physiology.

Hormonal daily variation co-varies with immunity in captive male bullfrogs (Lithobates catesbeianus)

Almost all physiological processes within the organism, including immune parameters and hormones, follow a circadian rhythm. These daily fluctuations are often observed in free-living organisms; however, little is known regarding hormonal and immune daily variations in anurans, particularly under laboratory conditions. This study aimed to investigate the hormonal and immune daily variation in captive-bred Bullfrogs (Lithobates catesbeianus) under constant conditions (21 °C and 12:12 LD cycle). Our results showed a daily variation for plasma corticosterone (CORT), testosterone (T), and melatonin (MEL), as well as for blood leukocyte profile, phagocytic activity, and plasma bacterial killing ability (BKA). Hormonal profile and immune activity were higher at the dark when compared with the light phase; however, monocytes and lymphocytes followed the opposite pattern. Moreover, CORT was positively correlated with phagocytosis percentage of blood cells, BKA, and monocytes, while MEL and T showed a positive correlation with PP. Our results demonstrate the daily covariation of different immune variables and immunomodulatory hormones. These 24 h-day variations and covariation certainly have broad implications and need to be considered for better understanding anuran physiology both in the context of laboratory and field studies.

Systemic hormonal and immune regulation induced by intraperitoneal LPS injection in bullfrogs (Lithobates catesbeianus)

Host's defense against external challenges activates an inflammatory response regulated by a set of chemical signals, including hormones. These immunomodulatory hormones, such as corticosterone, testosterone, and melatonin, trigger the systemic immune responses responsible for inflammatory assembly and resolution. This study aimed to investigate the effects of an immune challenge on endocrine and innate immune responses in the bullfrog (Lithobates catesbeianus). Adult males were intraperitoneally injected with lipopolysaccharide (LPS; 2 mg/kg) or saline, and blood samples were collected 6 and 24 h after injection for measurement of neutrophil/lymphocyte ratio, blood leukocyte phagocytosis, plasma bacterial killing ability, and plasma levels of corticosterone, melatonin, and testosterone. Our results showed LPS-induced increased neutrophil/lymphocyte ratio and leukocyte phagocytosis, and decreased melatonin and testosterone plasma levels, which were more pronounced 24 h after injection. Overall, we conclude that LPS intraperitoneal injection can activate the innate immune response and modulate the hormonal profile of the bullfrogs, with effects more pronounced 24 h than 6 h after treatment.

Challenges of a novel range: Water balance, stress, and immunity in an invasive toad

Species introduced by human activities can alter the normal functioning of ecosystems promoting negative impacts on native biodiversity, as they can rapidly expand their population size, demonstrating phenotypic plasticity and possible adaptive capacity to novel environments. Twenty years ago, the guttural toad, Sclerophrys gutturalis, was introduced to a peri-urban area of Cape Town, with cooler and drier climatic characteristics than its native source population, Durban, South Africa. Our goal was to understand the phenotypic changes, in terms of physiology and immunity, of populations in native and novel environments. We evaluated body index (BI), field hydration level, plasma corticosterone levels (CORT), proportion of neutrophils: lymphocytes (N: L), plasma bacterial killing ability (BKA), and hematocrit (HTC) in the field, and after standardized stressors (dehydration and movement restriction) in males from the native and invasive populations. Toads from the invasive population presented lower BI and tended to show a lower field hydration state, which is consistent with living in the drier environmental conditions of Cape Town. Additionally, invasive toads also showed higher BKA and N:L ratio under field conditions. After exposure to stressors, invasive animals presented higher BKA than the natives. Individuals from both populations showed increased CORT after dehydration, an intense stressor for these animals. The highest BKA and N:L ratio in the field and after submission to stressors in the laboratory shows that the invasive population has a phenotype that might increase their fitness, leading to adaptive responses in the novel environment and, thus, favoring successful dispersion and population increase.

Long-term monitoring of two snake species reveals immune-endocrine interactions and the importance of ecological context.

While there is huge promise in monitoring physiological parameters in free-living organisms, we also find high amounts of variability over time and space. This variation requires us to capitalize on long-term physiological monitoring to adequately address questions of population health, conservation status, or evolutionary trends as long-term sampling can examine ecoimmunological and endocrine interactions in wild populations while accounting for the variation that often makes ecophysiological field studies difficult to compare. In this study, we tested how immune efficacy and endocrinology interact while accounting for ecological context and environmental conditions in two snake species. Specifically, we measured bacterial killing ability, steroid hormones, and morphological characteristics in multiple populations of the Western Terrestrial Gartersnake (Thamnophis elegans) and Common Gartersnake (T. sirtalis) for multiple seasons over 6 years. Leveraging this long-term dataset, we tested how a broad immune measure and endocrine endpoints interact while accounting for individual traits, sampling date, and environmental conditions. Across both species, we found bacterial killing ability to be directly related to corticosterone(CORT) and temperature and greater overall in the spring compared to the fall. We found CORT and testosterone yielded relationships with individual sex, sampling temperature, and time of year. Wild populations can exhibit high amounts of variation in commonly collected physiological endpoints, highlighting the complexity and difficulty inherent in interpreting single endpoints without taking ecological and environmental conditions into account. Our study emphasizes the importance of reporting the environmental conditions under which the sampling occurred to allow for better contextualization and comparison between studies.

Thermal sensitivity of Bullfrog's immune response kept at different temperatures.

Climate change and emerging infectious diseases are often described as the main factors associated with the worldwide amphibian population decline. In this context, rising temperatures due to global warming might act as a chronic stressor for many amphibians, leading to immunosuppression. This study aimed to characterize the thermal sensitivity of the Bullfrog's (Lithobates catesbeianus) immune response and the effect of acclimation at different temperatures on it. Plasma bacterial killing ability (BKA) and phagocytosis activity of blood leukocytes were measured at different incubation temperatures (5-40°C) in individuals kept at 28°C and 34°C. First, all individuals were held under 28°C and sampled on the 16thday. Subsequently, one group was kept at 28°, and the other one was transferred to 34°C. Both groups were sampled at 83 and 106 days of maintenance. Plasma corticosterone (CORT) and testosterone (T) were assessed to evidence thermal stress and possible endocrine correlates of immune changes over time. The incubation temperature affected BKA both on animals kept at 28°C and 34°C, with maximum values at lower temperatures (5-20°C). Phagocytosis activity was constant over the range of assay temperatures. Immune and endocrine variables decreased over time in both thermal regimes, but frogs maintained at 34°C showed lower T and immunosuppression, evidencing stress response. Therefore, exposure to high temperatures might decrease immune function in bullfrogs due to chronic stress response and by exposition to temperatures of lower performance according to the thermal sensitivity curve, which might increase vulnerability to diseases in this anuran species.

Lipoxin A4 promotes reduction and antibiotic efficacy against Pseudomonas aeruginosa biofilm

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic bacterium commonly found in wound infections and airways of cystic fibrosis patients. P. aeruginosa readily forms biofilms which can reduce the efficacy of antibiotics used to eradicate the pathogen. We have previously shown that a Specialized Pro-resolving Mediator (SPM), Lipoxin A4 (LxA4) is a quorum sensing inhibitor which can reduce P. aeruginosa virulence. In this study, we examined the direct actions of LxA4 and RvD2 on P. aeruginosa biofilm formation and virulence gene expression. The influence of LxA4 on antibiotic efficacy and the combined effects on biofilm formation were also investigated. LxA4 and RvD2 reduced P. aeruginosa biofilm formation and virulence gene expression. LxA4 increased ciprofloxacin inhibition on biofilm formation but did not affect ciprofloxacin’s action on non-adherent bacteria. On the other hand, LxA4 increased bacterial killing action of imipenem but did not affect imipenem’s action on biofilm. We also found that LxA4 can increase ciprofloxacin’s bacterial killing ability in established biofilm. Together these results suggest that LxA4 has direct effects on P. aeruginosa biofilm formation and can increase antibiotic efficacy directly.

Competing reproductive and physiological investments in an all-female lizard, the Colorado checkered whiptail

Organisms in the wild have to allocate limited resources towards competing functions such as reproduction, growth, and self-maintenance. These competing investments create significant changes in physiological activity, and we still know little about the relationship between physiological activity and reproductive investment in natura. We investigated trade-offs between physiological activity and reproductive investment in the parthenogenetic Colorado checkered whiptail, Aspidoscelis neotesselata, across three different sites at the US Army Fort Carson Military Installation near Colorado Springs, CO, throughout the reproductive season in 2018 and 2019. We measured clutch size and reproductive activity and quantified plasma corticosterone (CORT), reactive oxygen metabolites (ROMs), and bacterial killing ability (BKA) to examine how energy-mobilizing hormones, oxidative stress, and immunity change in light of reproductive investment across different sub-populations. Circulating CORT increased with reproductive investment across all sub-populations, and increased clutch size led to a decrease in BKA in one sub-population, suggesting that habitat and nutritional availability may mediate this relationship. Oxidative stress, CORT, and innate immunity were not correlated with the exception of a trade-off between ROMs and BKA. This indicates individuals that have a better capacity to fight-off pathogens suffered increased reactive oxygen metabolites across all sub-populations, independently of habitat characteristics, which has important implications for A. neotesselata conservation.

Glucocorticoids, energy metabolites, and immunity vary across allostatic states for plateau side-blotched lizards (Uta stansburiana uniformis) residing in a heterogeneous thermal environment.

Reptiles rely on thermal heat exchange to achieve body temperatures (Tbody ) conducive to maintaining homeostasis. Diurnal changes in the thermal environment are therefore liable to influence allostatic mediation of survival processes (e.g., immunity) during environmental challenges or stressors. However, the extent to which Tbody prompts individual variation in physiology remains largely unexplored in reptiles. Our study tested how circulating energy-mobilizing hormone, energy metabolites, and immunity can vary across basal and stress-induced allostatic states for plateau side-blotched lizards (Uta stansburiana uniformis) residing in a heterogeneous thermal environment. We collected baseline and acute stress blood samples from male lizards to compare changes in plasma corticosterone (CORT), glucose, and bacterial killing ability (BKA) in relation to each other and Tbody . We hypothesized each physiological parameter differs between allostatic states, whereby stress-induced activity increases from baseline. At basal and stress-induced states, we also hypothesized circulating CORT, glucose, and BKA directly correspond with each other and Tbody . We found both CORT and BKA increased while glucose instead decreased from acute stress. At basal and stress-induced allostatic states, we found CORT to be directly related to Tbody while BKA was inversely related to CORT. We also found BKA and glucose were directly related at baseline, but inversely related following acute stress. Overall, these results demonstrate allostatic outcomes from acute stress in a free-living reptile and the role of temperature in mediating energetic state and immunity. Future research on reptilian allostasis should consider multiple environmental conditions and their implications for physiological performance and survival.

Polynorepinephrine as an Efficient Antifouling-Coating Material and Its Application as a Bacterial Killing Photothermal Agent

Biomedical device-related infection (BDI) is of great concern in modern clinical and medical applications. Various approaches to combat BDI are based on two major principles: the prevention of biofoulants adhering on medical devices and the ability to eradicate biofouling once formed. To minimize the risk of BDI, an antifouling coating with bactericidal ability is highly desirable. In this work, we report on the use of polynorepinephrine (PNE) as a promising strategy to prevent BDI due to its excellent antifouling and photothermal bacterial killing capabilities. PNE coatings show superior protein resistance against a model biofoulant (bovine serum albumin (BSA)) when compared with poly(ethylene glycol) (PEG) and polydopamine (PDA) coatings. The antifouling mechanism between BSA protein molecules and coating films is investigated using atomic force microscopy (AFM). We also demonstrate that PNE-modified surfaces show remarkable bacterial killing ability against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria after being irradiated with an 850 nm near-infrared (NIR) laser. These results indicate that PNE coatings present a highly promising candidate for biomedical antifouling applications.