Bacterial Load Research Articles

Microbial metabolism disrupts cytokine activity to impact host immune response

Host–pathogen interactions are shaped by the metabolic status of both the host and pathogen. The host must regulate metabolism to fuel the immune response, while the pathogen must extract metabolic resources from the host to enable its own survival. In this study, we focus on the metabolic interactions of Mycobacterium abscessus with Drosophila melanogaster . We identify MAB_1132c as an asparagine transporter required for pathogenicity in M. abscessus . We show that this requirement is specifically associated with damage to the host: flies infected with MAB_1132c knockout bacteria, or with wild-type bacteria grown in asparagine-restricted conditions, are longer lived without showing a significant change in bacterial load. This is associated with a reduction in the host innate immune response, demonstrated by the decreased transcription of antimicrobial peptides as well as a significant reduction in the ability of the infection to disrupt systemic insulin signaling. Much of the increase in host survival during infection with asparagine-limited M. abscessus can be attributed to alterations in unpaired cytokine signaling. This demonstrates that asparagine transport in M. abscessus prior to infection is not required for replicative fitness in vivo but does significantly influence the interaction with the host immune responses.

Evaluation of LPRDA Pentapeptide for the Prevention and Treatment of Staphylococcus aureus Peritoneal Infection

Targeting virulence determinants is a promising approach to controlling S. aureus infections in the face of the global spread of antibiotic resistance. S. aureus-induced peritonitis often occurs in dialysis, implant and trauma patients. To develop novel prevention and treatment options for peritoneal infection, we investigated the oligopeptide sortase A inhibitor LPRDA as a non-conventional antibacterial that does not affect staphylococcal survival. Administration of LPRDA prior to S. aureus challenge reduced the bacterial load of internal organs and bacterial colonization of the abdominal cavity in animals. In addition, LPRDA inhibited α-hemolysin production in 80% of the 35 reference and clinical S. aureus strains tested. Consequent research of LPRDA interactions with cefazolin and vancomycin has demonstrated the potential for combined application of the antivirulent and antibiotic agents under study.

Unraveling Water-Soluble Constituents of Basil (Ocimum basilicum L.) in Relation to Their Toxicity and Anti-Typhoidal Activity in Mouse Models.

Plants are the major source of natural flavour ingredients reported for their wide applications in food and pharmaceuticals, oral care and wellness products, etc. We have investigated the water-soluble fractions (WSF) of basil tetraploid (O. basilicum L.) for their toxicity and biological potential against Salmonella Typhimurium, a pathogen causing around one million cases of illnesses in the United States every year. The WSF obtained using a Clevenger-type apparatus was further divided into two equal parts, one each for in-vivo toxicity evaluation and quality assessments, respectively. The proportions of major phenylpropanoid identified as meta-eugenol in the WSF were found in the range of 42.8-57.9%, which was substantially in higher proportion as compared to essential oil (20.9-23.0%). Based on sub-acute oral toxicity data, WSF has not shown any adverse effect with levels as high as 500 µL/25g body weight in Swiss albino mice. Besides, the WSF also exhibited a maximum reduction in bacterial load in mice infected with Salmonella Typhimurium in a dose-dependent manner. We have shown the biological potential of basil water-soluble fraction as an effective bacterial load-suppressing agent for the prevention of Salmonella infections in animal model.

Addressing underestimation of waterborne disease risks due to fecal indicator bacteria bound in aggregates.

This study aims to identify and address significant limitations in current culture-based regulatory methods used for monitoring microbiological water quality. Specifically, these methods' inability to distinguish between planktonic forms and aggregates containing higher bacterial loads and associated pathogens may lead to a severe underestimation of exposure risks, with critical public health implications. We employed a novel methodology combining size fractionation with ALERT, an automated rapid method for comprehensive quantification of culturable fecal indicator bacteria (FIB). Our findings reveal a substantial and widespread presence of aggregate-bound indicator bacteria across various water matrices and geographical locations. Comprehensive bacterial counts consistently exceeded those obtained by traditional methods by significant multiples, such as an average of 3.4×at the Seine River 2024 Olympic venue, and occasionally up to 100×in irrigation canals and wastewater plant effluent. These results, supported by microscopic and molecular analyses, underscore a systematic bias in global water safety regulatory frameworks. Our research demonstrates the inadequacy of traditional culture-based techniques in assessing microbiological risks posed by aggregate-bound FIB and associated pathogens, particularly in water matrices affected by FIB-rich fecal particles from recent sewer overflows or sediment, which can carry higher infectious risks. Incorporating comprehensive FIB analysis techniques, including molecular methods and rapid culture-based approaches as shown in this study, offers a promising and effective solution to these risk assessment limitations.

Bacterial contamination of autologous blood salvaged during deceased donor liver transplantation: a prospective observational study.

Decompensated cirrhotic patients experience severely increased intestinal permeability and bacterial translocation. Thus, autologous blood salvaged during deceased donor liver transplantation (DDLT) may be contaminated with enteric bacteria. We aimed to evaluate bacterial contamination of autologous blood salvaged during DDLT and its association with post-transplant bacteremia. In 30 patients undergoing DDLT, bacterial culture was performed in salvaged autologous blood samples: one before graft reperfusion (non-leukoreduced) and two after graft reperfusion (non-leukoreduced and leukoreduced). The primary outcome was bacterial contamination of salvaged autologous blood. Seven of 30 patients (23.3%) were positive for bacteria (3 enteric/4 non-enteric) before graft reperfusion while 11 patients (36.7%) were positive (5 enteric/6 non-enteric) after graft reperfusion. Six of 7 patients who were positive for bacteria before graft reperfusion were positive after graft reperfusion with the same bacteria. Only 4 of 11 contaminated blood samples were converted to negative after leukoreduction. Post-transplant bacteremia risk was insignificantly greater in patients who received autologous blood with bacteria than in patients without bacteria (30.0% vs. 5.0%, P = 0.06). We found contamination of salvaged autologous blood with enteric bacteria throughout DDLT and incomplete performance of leukoreduction, indicating high bacterial load. The potential association between contaminated autotransfusion and post-transplant bacteremia warrants further validation in a larger prospective study.Clinical trial notation: This study was registered at the Clinical Research Information Service (CRiS; https://cris.nih.go.kr ; No. KCT0007223; principal registration investigator: Sangbin Han, date of registration: April 25, 2022).

Indoor air bacterial quality and associated factors in prison inmate cells of East Hararghe Zone and Harari Regional State, Eastern Ethiopia.

Bacterial indoor air load refers to the level of bacteria within and around dwellings and other structures. Pathogens, bacterial cell fragments, and bacterial organisms' byproducts can all pose major issues indoors, especially in prison inmate cells. However, there is lack of data on bacterial load and contributing factors in the East Hararghe zone and Harari regional state. The lack of studies on microbiological indoor air quality in prisons with contributing factors will therefore be filled by this investigation. The study aimed to assess bacterial indoor air load and contributing factors in prison inmate cells from October 1 to October 30, 2020. An institutional cross-sectional study was employed. All of the prisons in the East Hararghe zone and the Harari regional state served as the study's and source population. 62 prison cells were used in the investigation. Samples were obtained using the passively settling plate technique. The data were evaluated through the use of SPSS statistical software, Excel, and the statistical procedures of ANOVA, correlation, and chi-square test. The maximum and minimum bacterial loads, were recorded at 8:00 am (3027 CFU/m3) and 2:00 pm (1048 (CFU/m3) respectively. The correlation between the temperature and bacterial load was strongly positive (r = 0.680, p = 0.047), and the correlation of the moisture content and bacterial load was strongly negative (r = -0.671, p = 0.039). The levels of bacteria were higher than the guideline (2000 CFU/m3). While the relative humidity of indoor air was negatively correlated with bacterial load, temperature and bacterial load were significantly positively correlated. Harari regional state and East Hararghe zone prison commissions should be alarmed to alleviate these problems. The building standards need to be completely updated to the latest standards.

Ivacaftor ameliorates mucus burden, bacterial load, and inflammation in acute but not chronic P. aeruginosa infection in hG551D rats

BackgroundNewly approved highly effective modulation therapies (HEMT) have been life-changing for people with CF. Although these drugs have resulted in significant improvements in lung function and exacerbation rate, bacterial populations in the lung have not been eradicated. The mechanisms behind the continued colonization are not completely clear.MethodsWe used a humanized rat to assess the effects of ivacaftor therapy on infection outcomes. Rats harbor an insert expressing humanized CFTR cDNA, including the G551D mutation. hG551D rats were treated with ivacaftor either during or before infection with P. aeruginosa. The response to infection was assessed by bacterial burden in the lung and mucus burden in the lung.ResultsWe found that hG551D rats treated with ivacaftor had reduced bacteria present in the lung in the acute phase of the infection but were not different than vehicle control in the chronic phase of the infection. Similarly, the percentage of neutrophils in the airways were reduced at the acute, but not chronic, timepoints. Overall weight data indicated that the hG551D rats had significantly better weight recovery during the course of infection when treated with ivacaftor. Potentiation of the G551D mutation with ivacaftor resultant in short-circuit current measurements equal to WT, even during the chronic phase of the infection. Despite the persistent infection, hG551D rats treated with ivacaftor had fewer airways with mucus plugs during the chronic infection.ConclusionsThe data indicate that the hG551D rats have better outcomes during infection when treated with ivacaftor compared to the vehicle group. Rats have increased weight gain, increased CFTR protein function, and decreased mucus accumulation, despite the persistence of infection and inflammation. These data suggest that ivacaftor improves tolerance of infection, rather than eradication, in this rat model.

Activity of Bambara Groundnut Seed Coat Extract Against Shewanella Species: Efficacy and Mechanisms of Action

The Bambara groundnut is the indigenous legume in the southern part of Thailand. It contains a seed coat rich in polyphenols, which can serve as natural antimicrobial agents. The extracts from red and white seed coats of Bambara groundnuts, namely RSC and WSC, respectively, were prepared using an ultrasound-assisted extraction process. The extraction yield, total phenolic content (TPC), and antimicrobial activity of both extracts were examined. The RSC extract demonstrated a significantly higher extraction yield (8.35%) than WSC extract (2.34%) (p < 0.05). Furthermore, the TPC of RSC extract (420.98 ± 0.27 mg of gallic acid/g dry extract) was higher than that of WSC extract (28.29 ± 0.91 mg of gallic acid/g dry extract). The RSC extract exhibited stronger inhibition against Shewanella putrefaciens and S. algae than its WSC counterpart. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) analysis indicated that the RSC extract was rich in flavonoids and polyphenols, while the WSC extract contained more triterpenoid saponins. Time–kill kinetics showed that the RSC extract reduced bacterial loads in a dose-dependent manner. Scanning electron microscopic images revealed that drastic bacterial cell membrane damage with a rough surface and the deformation of cells was caused by the extract. Furthermore, confocal laser scanning microscopic (CLSM) images confirmed the inhibition of S. algae biofilm formation by RSC extract. RSC extract also suppressed bacterial motility, induced protein leakage, and reduced extracellular protease activity, thus highlighting its potent bactericidal effects. These findings suggested that the RSC extract rich in phenolic compounds could serve as an antimicrobial agent and hold promise as a natural preservative for perishable foods, especially seafoods.

Tissue nano-transfection of antimicrobial genes drives bacterial biofilm killing in wounds and is potentially mediated by extracellular vesicles

The emergence of bacteria that are resistant to antibiotics is on track to become a major global health crisis. Therefore, there is an urgent need for new treatment options. Here, we studied the implementation of tissue-nanotransfection (TNT) to treat Staphylococcus aureus-infected wounds by delivering gene cargos that boost the levels of naturally produced antimicrobial peptides. The Cathelicidin Antimicrobial Peptide gene (CAMP), which produces the antimicrobial peptide LL-37, was used as model gene cargo. In vitro evaluation showed successful transfection and an increase in the transcription and translation of CAMP-coding plasmid in mouse primary epithelial cells. Moreover, we found that the extracellular vesicles (EVs) derived from the transfected cells (in vitro and in vivo) carried significantly higher concentrations of CAMP transcripts and LL-37 peptide compared to control EVs, possibly mediating the trafficking of the antimicrobial contents to other neighboring cells. The TNT platform was then used in vivo on an excisional wound model in mice to nanotransfect the CAMP-coding plasmid on the edge of infected wounds. After 4 days of daily treatment, we observed a significant decrease in the bacterial load in the CAMP-treated group compared to the sham group. Moreover, histological analysis and bacterial load quantification also revealed that TNT of CAMP on S. aureus-infected wounds was effective in treating biofilm progression by reducing the bacterial load. Lastly, we observed a significant increase in macrophage recruitment to the infected tissue, a robust increase in vascularization, as well as and an increased expression of IL10 and Fli1. Our results demonstrate that TNT-based delivery of gene cargos coding for antimicrobial compounds to the wound is a promising approach for combating biofilm infections in wounds.

Refining the gut colonization Zophobas morio larvae model using an oral administration of multidrug-resistant Escherichia coli

The darkling beetle Zophobas morio can be implemented as an alternative in vivo model to study different intestinal colonization aspects. Recently, we showed that its larvae can be colonized by multidrug-resistant Escherichia coli strains administered via contaminated food (for 7 days) for a total experimental duration of 28 days. In the present work, we aimed to shorten the model to 14 days (T14) by administering the previously used CTX-M-15 ESBL-producing ST131 Escherichia coli strain Ec-4901.28 via a single oral administration (5 µL dose of 108 CFU/mL) , using a blunt 26s-gauge needle connected to a 250 μL gastight syringe. Force-feeding was performed either without or with (larvae placed on ice for 10 minutes before injection) anesthesia. In addition, phage-treated larvae were orally injected with 10 µL of INTESTI bacteriophage cocktail (∼105-6 PFU/mL) on days 4 (T4) and 7 (T7) . Growth curve analyses showed that, while larvae rapidly became colonized with Ec-4901.28 (T1, ∼106-7 CFU/mL) , only those anesthetized maintained a high bacterial load (∼102-3vs. ∼105-6 CFU/mL) and survival rate (76% vs. 99%; P<0.001) by T14. Moreover, bacteriophage administration to anesthetized larvae significantly reduced the bacterial count of INTESTI-susceptible Ec-4901.28 at T14 (5.17 × 105vs. 2.26 × 104, for non-treated and phage-treated larvae, respectively; P=0.04) . The methodological refinements applied to establish the intestinal colonization model simplify the use of Z. morio larvae, facilitate prompt evaluation of novel decolonization approaches and reduce experiments involving vertebrate animals in accordance with the Replacement, Reduction and Refinement principles.

Leptospira Leptolysin Contributes to Serum Resistance but Is Not Essential for Acute Infection.

Previous invitro works focusing on virulence determinants of the spirochete Leptospira implicated metalloproteinases as putative contributing factors to the pathogenicity of these bacteria. Those proteins have the capacity to degrade extracellular matrix components (ECM) and proteins of host's innate immunity, notably effectors of the complement system. In this study, we gained further knowledge on the role of leptolysin, one of the leptospiral-secreted metalloproteinases, previously described as having a broad substrate specificity. We demonstrated that a proportion of human patients with mild leptospirosis evaluated in the current study produced antibodies that recognize leptolysin, thus indicating that the protease is expressed during host infection. Using recombinant protein and a knockout mutant strain, Manilae leptolysin-, we determined that leptolysin contributes to Leptospira interrogans serum resistance invitro, likely by proteolysis of complement molecules of the alternative, the classical, the lectin, and the terminal pathways. Furthermore, in a hamster model of infection, the mutant strain retained virulence; however, infected animals had lower bacterial loads in their kidneys. Further studies are necessary to better understand the role and potential redundancy of metalloproteinases on the pathogenicity of this important neglected disease.

Characterization of a Mycoplasma hyopneumoniae aerosol infection model in pigs

The purpose of the present study was to develop and characterize an experimental aerosol model for Mycoplasma hyopneumoniae (M. hyopneumoniae) infection and respiratory disease in pigs. The experiment was carried out to determine the pathogenicity, colonization, mucosal immune response, and clinical course of disease of dose-controlled aerosols of M. hyopneumoniae. Four groups of three M. hyopneumoniae-free gilts each were individually exposed to aerosols of diluted lung homogenate containing M. hyopneumoniae strain 232 in a chamber. Each group was exposed to different doses of viable organisms (105 to 106 color changing units/mL during 15-20 or 30-35min in two consecutive days). Nasal, laryngeal, and deep-tracheal secretions were collected from each gilt at 0, 7, 14, 21, and 28 days post-exposure (dpe). Blood samples were collected at 0 and 28 dpe. At necropsy, lung lesions were assessed, and bronchial secretions and bronchoalveolar lavage fluid (BALF) were collected from each lung set. Blood was used to assess seroconversion by means of an indirect ELISA, while BALF, deep-tracheal and nasal secretions were tested by modifying the ELISA to evaluate mucosal IgG and IgA production. Nasal, laryngeal, deep-tracheal, and bronchial secretions were tested by real-time PCR to evaluate bacterial load. Gilts became infected irrespective of the infectious dose, as determined by M. hyopneumoniae detection in deep-tracheal secretions from all gilts at 7 dpe. A specific local humoral immune response starting at 14 dpe was detected in all gilts. While all experimental groups presented gilts with some extent of mycoplasmal pneumonia, only the exposure of gilts to high-dose aerosols consistently reproduced typical clinical signs and severe lung lesions. These results showed that the reproduction of mycoplasmal pneumonia by means of infectious aerosols can be successfully achieved at experimental level, making this model a valuable alternative to evaluate preventive and treatment measures against M. hyopneumoniae.

Associative analysis of sludge microbiota and wastewater degradation efficacy within swine farm sludge systems

Industrial wastewater management is a significant global challenge. Sludge microbiota from swine farms may play a crucial role in enhancing wastewater treatment processes, thereby reducing water pollution from industrial activities. A deeper understanding of this complex community could lead to innovative approaches for improving wastewater treatment methods. Sludge samples were collected from the anaerobic, sedimentation, and thickening tanks of ten swine farms. The microbiota communities were analyzed using 16S rRNA full-length sequencing on the PacBio platform, with subsequent data analysis conducted on the QIIME2 platform utilizing the SILVA database. Compared to anaerobic and thickening tanks, the sedimentation tanks exhibited a unique profile of sludge microbiota, with higher abundances of the phyla Proteobacteria, Bacteroidota, and Caldatribacteriota. Additionally, sludges from farms already utilized in processing industrial water—specifically farms B, G, and J—contained higher concentrations of bacteria (> 20 ng/μL), indicating the robustness of the bacterial load for practical industrial use. Furthermore, sludge from farms with higher alpha diversity, such as E, G, I, and J, exhibited enriched degradation profiles, including the degradation of aromatic compounds, polymers, industrial compounds, toluene, and vanillin. The farms were categorized based on wastewater ammonia nitrogen degradation levels, revealing a clustering effect of the microbiota from the sedimentation tanks in the Principal Coordinates Analysis (PCoA) plot. A higher relative abundance of the families Rhodocyclaceae, AKYH767, and Comamonadaceae, and a lower abundance of the families Anaerolineaceae and Christensenellaceae, were found in groups with high ammonia nitrogen reduction, suggesting potential targets for bioaugmentation strategies. In conclusion, this study underscores the critical role of microbial abundance, composition, and biodiversity in optimizing wastewater treatment and advocates for comprehensive microbiota analysis to identify suitable sludge for industrial applications.

Development of α-Helical Antimicrobial Peptides with Imperfect Amphipathicity for Superior Activity and Selectivity.

The advancement of antimicrobial peptides (AMPs) as therapeutic agents is hindered by their poor selectivity. Recent evidence indicates that controlled disruption of the amphipathicity of α-helical AMPs may increase the selectivity. This study investigated the role of imperfect amphipathicity in optimizing AMPs with varied sequences to enhance their activity and selectivity. Among these, the lead peptide RI-18, characterized by an imperfectly amphipathic α-helical structure, demonstrated potent and broad-spectrum antibacterial activity without inducing hemolytic or cytotoxic effects. RI-18 effectively eliminated planktonic and biofilm-associated bacteria as well as persister cells and exhibited high bacterial plasma membrane affinity, inducing rapid membrane permeabilization and rupture. Notably, RI-18 significantly reduced bacterial loads without promoting bacterial resistance, highlighting its therapeutic potential. Overall, this study identified RI-18 as a promising antimicrobial candidate. The rational strategy of tuning imperfect amphipathicity to enhance the AMP activity and selectivity may facilitate the design and development of AMPs.

Effect of gamma radiation on sensory, microbial and lipid quality of whole Indian mackerel (Rastrelliger kanagurta)

The study focused on the impact of γ-radiation on the lipids, sensory attributes, and microbial quality of fresh whole mackerel. Fish were irradiated at 2, 4 and 6 kGy doses; subsequently, microbial load, sensory quality and fatty acid composition were studied. The results indicated no significant changes in sensory quality up to 6 kGy dose. The bacterial load of 4.75log CFU/g was reduced to 3.54, 2.74, 2.17log CFU/g at 2, 4 and 6 kGy doses respectively. The total mould count reduced from 1.17log CFU/g to 0.7log CFU/g at 2 kGy, and was below the detection limit at 4 and 6 kGy. No significant difference was observed in fatty acid composition at a dose of 2 kGy compared to control samples. However, a significant increase in saturated fatty acid and monounsaturated fatty acid with a significant decrease in polyunsaturated fatty acid content was detected at 4 and 6 kGy doses. Based on these findings, the study suggests that gamma irradiation at 4 kGy is effective for microbial safety with minimal impact on fatty acid composition.

Assessment and quantification of the microbial flora in the output water of two types of dental chairs: A comparative study

Objectives: Dental Unit Waterlines (DUWL) have shown to be a perfect host for different pathogenic microorganisms. The purpose of this study was to analyze and compare the level of bacterial contamination in the output water of 2 types of DUWL. Methods: Dental unit water samples from the air/water syringe of the A-dec (A-dec™ Performer 200, Newberg, USA) DUWL type and KaVo (ESTETICA™ E30/E70/E80 Vision, Kavo, Biberach, Germany) type were collected and analyzed for total aerobic flora, Pseudomonas aeruginosa, faecal coliforms, total coliforms, faecal streptococci and sulfite-reducing anaerobic flora. Results: The bacteriological analysis of water samples shows the presence of bacterial contamination at high levels exceeding the standard safety guidelines of 100 CFU/mL set up by the American Dental Association (ADA) and accepted by the Centers for Disease Control and prevention (CDC) on the heterotrophic bacterial load. The data shows no statistically significant differences for all bacteriological parameters studied between the conventional A-dec DUWL type and the KaVo type that has an automated mode of decontamination. Conclusions: Despite all the disinfecting solutions considered to eradicate the bacterial proliferation in DUWL, the problem remains one of the greatest challenges in modern dentistry. Practitioners and medical staff should not underestimate the harmful consequences of this bacterial growth, not only on the health of their patients but also on their own health.

The role of TF-b in iron homeostasis and bacterial defense in common carp (Cyprinus carpio)

Transferrin (TF) is a prototypical biological macromolecule protein known for its iron-binding properties. TF proteins play a crucial role in modulating host iron homeostasis and defending against pathogen invasion. In this study, we utilized common carp (Cyprinus carpio) tissues and Epithelioma papulosum cyprinid (EPC) cells to establish experimental models of iron overload with FeCl3 or ferric amine citrate (FAC), and to establish experimental models of bacterial infection with Aeromonas hydrophila. The current research has successfully identified the CcTF-b gene in common carp, revealing an ORF of 2001 bp with N-terminal and C-terminal structures. CcTF-b exhibited inhibitory effects on the growth of LPS and LTA in vitro. In the experimental models, the upregulations of PTGS2a and PTGS2a-like mRNA and protein levels were observed. Overexpression or interference with CcTF-b levels can modulate the expression of ferroptosis-related genes, inflammatory cytokines, lipid reactive oxygen species, GSH/GSSH levels, and Fe2+ concentration. Significantly, the expression levels of Nrf2 and GPX4 mRNA and protein, as well as the bacterial load of A. hydrophila, could be also modulated either by upregulating or downregulating CcTF-b factors. In conclusion, in this study, these findings suggest that CcTF-b plays a critical role in the innate immune response of common carp.

A Standardized Mouse Model for Wound Infection with Pseudomonas aeruginosa

Pseudomonas aeruginosa is a highly drug-resistant pathogen known to impair wound healing and provoke inflammatory responses, potentially leading to immune dysregulation. This study aimed to systematically investigate the immune response mechanisms mediated by cytokines following P. aeruginosa infection through the development of a standardized wound model. Kunming mice were selected as experimental subjects and given 8 mm diameter lesions on their backs and inoculated with standard strains PAO1 and PA14. The key parameters assessed included changes in body weight, wound redness and swelling, bacterial dynamics, protein content in wound tissues, immune responses, and pathological alterations. The results demonstrated that pathogen invasion significantly inhibited wound healing, with healing rates in the infected groups (87.5 ± 6.3% and 77.1 ± 3.6%) being notably lower than those in the uninfected control group. P. aeruginosa persisted in the wounds for up to 12 days, with bacterial loads decreasing from 8 log to 2 log. Additionally, there was a marked reduction in the protein content of the wound tissue and an increase in the expression levels of inflammatory factors such as IL-1β and TNF-α. The thickness of granulation tissue and the number of neovessels were significantly lower compared to the uninfected control group. This study establishes a standardized paradigm for creating a mouse model of P. aeruginosa infection in wounds, emphasizing the importance of appropriate mouse strains, uniform wound preparation methods, and moderate inoculation doses for reliable and accurate experimental results. These elements will facilitate the assessment of changes across six key indicators post-infection, providing a foundational data set and technical support for future mechanistic investigations of P. aeruginosa infection and the development of targeted antimicrobial strategies.

Hyaluronic acid methacrylate/Pluronic F127 hydrogel enhanced with spermidine-modified mesoporous polydopamine nanoparticles for efficient synergistic periodontitis treatment

Bacterial infection, reactive oxygen species (ROS) accumulation, and persistent inflammation pose significant challenges in the treatment of periodontitis. However, the current single-modal strategy makes achieving the best treatment effect difficult. Herein, we developed a double-network hydrogel composed of Pluronic F127 (PF-127) and hyaluronic acid methacrylate (HAMA) loaded with spermidine-modified mesoporous polydopamine nanoparticles (M@S NPs). The PF-127/HAMA/M@S (PH/M@S) hydrogel was injectable and exhibited thermosensitivity and photocrosslinking capabilities, which enable it to adapt to the irregular shape of periodontal pockets. In vitro, the PH/M@S displayed multiple therapeutic effects, such as photothermal antibacterial activity, a high ROS scavenging capacity, and anti-inflammatory effects, which are beneficial for the multimodal treatment of periodontitis. The underlying anti-inflammatory mechanism of this hydrogel involves suppression of the extracellular regulated protein kinase 1/2 and nuclear factor kappa-B signalling pathways. Furthermore, in lipopolysaccharide-stimulated macrophage conditioned media, the PH/M@S effectively restored the osteogenic differentiation potential. In a rat model of periodontitis, the PH/M@S effectively reduced the bacterial load, relieved local inflammation and inhibited alveolar bone resorption. Collectively, these findings highlight the versatile functions of the PH/M@S, including photothermal antibacterial activity, ROS scavenging, and anti-inflammatory effects, indicating that this hydrogel is a promising multifunctional filling material for the treatment of periodontitis.

Sustainable Bacterial Control of Hatching Eggshells Using Essential Oils

Background: Decontamination of poultry surfaces through appropriate hygiene and sanitation measures can partially mitigate bacterial problems, as this process does not result in the complete elimination of bacteria. Thus, the remaining bacteria can persist and contaminate eggshells. Therefore, regardless of the rigor of the sanitary standards applied on farms, it is suggested that hatching eggs be subjected to the sanitization process. Here, we investigated the effectiveness of essential oil-based antibacterial agents in sanitizing eggs. Results: The results indicated that essential oils from Cinnamomum cassia (L.) J. Presl. (CCEO), Syzygium aromaticum (L.) Merr. &amp; L.M. Perry (SAEO) and Cymbopogon nardus (L.) Rendle (CNEO), at specific concentrations, have antibacterial effects in vitro, reducing the load of mesophilic bacteria and enterobacteria in the eggshell by at least 3 and 2 log10 CFU/mL, respectively. Conclusion: The adoption of CCEO, SAEO and CNEO to reduce the bacterial load on eggshells could be a favorable change to the conventional protocol of egg sanitization with formaldehyde, especially on farms where sanitary standards are insufficient.