Pathogenic Bacteria Research Articles

Validation of Recombinase Polymerase Amplification with In-House Lateral Flow Assay for mcr-1 Gene Detection of Colistin Resistant Escherichia coli Isolates.

The emergence of the mobilized colistin resistance 1 (mcr-1) gene, which causes colistin resistance, is a serious concern in animal husbandry, particularly in pigs. Although antibiotic regulations in many countries have prohibited the use of colistin in livestock, the persistence and dissemination of this plasmid-mediated gene require effective and rapid monitoring. Therefore, a rapid, sensitive, and specific method combining recombinase polymerase amplification (RPA) with an in-house lateral flow assay (LFA) for the mcr-1 gene detection was developed. The colistin agar test and broth microdilution were employed to screen 152 E. coli isolates from pig fecal samples of five antibiotic-used farms. The established RPA-in-house LFA was validated with PCR for mcr-1 gene detection. The RPA-in-house LFA was completed within 35 min (20 min of amplification and 5-15 min on LFA detection) at 37 °C. The sensitivity, specificity, and accuracy were entirely 100% in concordance with PCR results. No cross-reactivity was detected with seven common pathogenic bacteria or other mcr gene variants. Therefore, the in-house RPA-LFA serves as a point-of-care testing tool that is rapid, simple, and portable, facilitating effective surveillance of colistin resistance in both veterinary and clinical settings, thereby enhancing health outcomes.

Effect of Dietary Benzoic Acid and Oregano Essential Oil as a Substitute for an Anti-Coccidial Agent on Growth Performance and Physiological and Immunological Responses in Broiler Chickens Challenged with Eimeria Species.

To overcome the antimicrobial residues in food, benzoic acid (BA) and oregano essential oil (OEO) are used in the broiler chicken industry. Independently, both exerted anticoccidial and antimicrobial actions and improved growth performance in broiler chickens. Their effect may be multiplied when they are used in combination. This present study was carried out to evaluate the efficacy of dietary BA and OEO alone or in combination as a substitute for a commercial coccidiostatic drug on growth performance and physiological and immunological responses in broiler chickens challenged with Eimeria species. A total of 252 unsexed 1-day-old broiler chicks were equally allotted to 36 pens, each pen containing seven chicks. The pens were randomly assigned to six treatments with six pens (replicates) for each treatment (n = 6)-(i) negative control, (ii) positive control, coccidia-challenged and non-treated, (iii) supplemented with salinomycin (an anti-coccidial drug) at 60 mg/kg of feed and coccidia-challenged, (iv) supplemented with BA at 500 mg/kg of feed and coccidia-challenged, (v) supplemented with OEOat 500 mg/kg of feed and coccidia-challenged (OEO), and (vi) supplemented with BA at 500 mg/kg of feed and OEO at 500 mg/kg of feed and coccidia-challenged (B&O). The liver enzymes and thyroxine and creatinine levels were not affected (p > 0.05) both in coccidia-challenged and supplemented chickens. The BA and OEO applied separately or in combination (B&O) significantly (p < 0.05) reduced gut pathogenic bacteria (Salmonella and Escherichia coli) and Eimeria spp., and concurrently enhanced (p > 0.05) the Lactobacillus population with better body weight gain, improved feed utilization, and superior hematological values. It also up-regulated (p > 0.05) the interferon-γ gene expression and down-regulated (p < 0.05) the interleukin-10 and Toll-like receptor-4 gene expression to protect the chickens from inflammatory reactions, which were not demonstrated in salinomycin-treated birds. The B&O supplementation increased (p < 0.05) the immune system by enhancing Eimeria-specific immunoglobulin Y titer and lymphocyte proliferation response. This study suggests that the combined application of OEO and BA can substitute for a commercial anti-coccidial agent (salinomycin) in controlling coccidiosis as well as improving growth performance, gut health, and immune responses in broiler chickens with a means of antimicrobial-resistant free food products.

Compressed sensing based approach identifies modular neural circuitry driving learned pathogen avoidance.

An animal's survival hinges on its ability to integrate past information to modify future behavior. The nematode C. elegans adapts its behavior based on prior experiences with pathogen exposure, transitioning from attraction to avoidance of the pathogen. A systematic screen for the neural circuits that integrate the information of previous pathogen exposure to modify behavior has not been feasible because of the lack of tools for neuron type specific perturbations. We overcame this challenge using methods based on compressed sensing to efficiently determine the roles of individual neuron types in learned avoidance behavior. Our screen revealed that distinct sets of neurons drive exit from lawns of pathogenic bacteria and prevent lawn re-entry. Using calcium imaging of freely behaving animals and optogenetic perturbations, we determined the neural dynamics that regulate one key behavioral transition after infection: stalled re-entry into bacterial lawns. We find that key neuron types govern pathogen lawn specific stalling but allow the animal to enter nonpathogenic E. coli lawns. Our study shows that learned pathogen avoidance requires coordinated transitions in discrete neural circuits and reveals the modular structure of this complex adaptive behavioral response to infection.

Identification and Assessment of Secondary Metabolites from Three Fungal Endophytes of Solanum mauritianum Against Public Health Pathogens

Fungal endophytes, symbiotic microorganisms residing within plants, are renowned for producing bioactive secondary metabolites with diverse beneficial properties. We investigated the antimicrobial potential of fungal endophytes isolated from Solanum mauritianum, an invasive weed, against clinically significant bacterial pathogens. Selected fungal endophytes (Penicillium chrysogenum, Fusarium sp., and Paracamarosporium leucadendri) were isolated from the plant’s leaves and fruits. Their crude extracts were tested against various referenced strains, such as Mycobacterium species (M. smegmatis ATCC 607 and M. bovis ATCC 27290), Staphylococcus aureus ATCC 6571, Bacillus subtilis ATCC 11774, Klebsiella species (K. pneumoniae ATCC 10031 and K. oxytoca ATCC 8724), Escherichia coli ATCC 10536, and Pseudomonas aeruginosa ATCC 10145, using the Kirby-Bauer disk diffusion method. Resazurin Microtiter Assay was used for the determination of the minimum inhibitory concentration. The chemical nature of the secondary metabolites in the crude extracts produced by fungal endophytes was evaluated using high-resolution liquid chromatography–mass spectrometry (LC-MS) using water and acetonitrile gradient. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS) was employed for untargeted metabolomics. LC-QTOF-MS/MS identified 63 bioactive compounds across the three endophytes. P. chrysogenum had the highest activity against S. aureus and M. smegmatis (1.15 mg/mL and 0.02 mg/mL, respectively), while P. leucadendri demonstrated moderate activity against M. smegmatis (2.91 mg/mL) and E. coli (1.16 mg/mL). Fusarium sp. exhibited the broadest spectrum of antibacterial activity, with MIC values ranging from 0.03 mg/mL (B. subtilis) to 10 mg/mL (M. smegmatis). P. leucadendri produced 29 metabolites, Fusarium sp. had 23 identified metabolites, and a total of 11 metabolites were identified from P. chrysogenum. The fruits of the plant, accounting for 60%, appeared to be the most abundant in the endophyte diversity when compared to the stems and leaves. This study highlights the potential of fungal endophytes from S. mauritianum as a source of novel bioactive compounds, particularly against multidrug-resistant pathogens, contributing to the ongoing efforts to combat antimicrobial resistance.

Altering the redox status of Chlamydia trachomatis directly impacts its developmental cycle progression.

Chlamydia trachomatis is an obligate intracellular bacterial pathogen with a unique developmental cycle. It differentiates between two functional and morphological forms: elementary body (EB) and reticulate body (RB). The signals that trigger differentiation from one form to the other are unknown. EBs and RBs have distinctive characteristics that distinguish them, including their size, infectivity, proteome, and transcriptome. Intriguingly, they also differ in their overall redox status as EBs are oxidized and RBs are reduced. We hypothesize that alterations in redox may serve as a trigger for secondary differentiation. To test this, we examined the function of the primary antioxidant enzyme alkyl hydroperoxide reductase subunit C (AhpC), a well-known member of the peroxiredoxins family, in chlamydial growth and development. Based on our hypothesis, we predicted that altering the expression of ahpC would modulate chlamydial redox status and trigger earlier or delayed secondary differentiation. To test this, we created ahpC overexpression and knockdown strains. During ahpC knockdown, ROS levels were elevated, and the bacteria were sensitive to a broad set of peroxide stresses. Interestingly, we observed increased expression of EB-associated genes and concurrent higher production of EBs at an earlier time in the developmental cycle, indicating earlier secondary differentiation occurs under elevated oxidation conditions. In contrast, overexpression of AhpC created a resistant phenotype against oxidizing agents and delayed secondary differentiation. Together, these results indicate that redox potential is a critical factor in developmental cycle progression. For the first time, our study provides a mechanism of chlamydial secondary differentiation dependent on redox status.

Translational research priorities for bacteriophage therapeutics.

The growing threat of antimicrobial resistant (AMR) bacterial pathogens coupled with the relative dearth of promising novel antibiotics requires the discovery and development additional medical interventions. Over the past decade bacteriophages have emerged one of the most promising new tools to combat AMR pathogens. Anecdotal clinical experiences under so-called 'compassionate use' regulatory pathways as well as a limited number of clinical trials have provided ample evidence of safety and early evidence of efficacy. For phages to reach their full potential it is critical that rigorous clinical trials be conducted that define their optimal use and that enable regulatory authorities to support the commercialization required to afford global access. The clinical development of phage therapeutics requires the design and execution of clinical trials that take full advantage of lessons learned from a century of antibiotic development and that use clinical investigation as a platform in which aspects of phage biology that are critical to therapeutics are more clearly elucidated. Translational research that elucidates phage biology in the context of clinical trials will promote highly relevant hypothesis-driven work in basic science laboratories and will greatly accelerate the development of the field of phage therapeutics.

Cold-water coral mortality under ocean warming is associated with pathogenic bacteria

Cold-water corals form vast reefs that are highly valuable habitats for diverse deep-sea communities. However, as the deep ocean is warming, it is essential to assess the resilience of cold-water corals to future conditions. The effects of elevated temperatures on the cold-water coral Lophelia pertusa (now named Desmophyllum pertusum) from the north-east Atlantic Ocean were experimentally investigated at the holobiont level, the coral host, and its microbiome. We show that at temperature increases of + 3 and + 5 °C, L. pertusa exhibits significant mortality concomitant with changes in its microbiome composition. In addition, a metagenomic approach revealed the presence of gene markers for bacterial virulence factors suggesting that coral death was due to infection by pathogenic bacteria. Interestingly, different coral colonies had different survival rates and, colony-specific microbiome signatures, indicating strong colony-specific variability in their response to warming waters. These results suggest that L. pertusa can only survive a long-term temperature increase of < 3 °C. Therefore, regional variations in deep-sea temperature increase should be considered in future estimates of the global distribution of cold-water corals.

Zoonotic Pathogens Isolated from an Introduced Population of Red Swamp Crayfish (Procambarus clarkii) in Tenerife (Canary Islands, Spain)

The red swamp crayfish (Procambarus clarkii) is a widely distributed invasive species that is listed in the Delivering Alien Invasive Species Inventory for Europe. Native to North America, it has been introduced to numerous regions, such as the Canary Islands, Spain. Previous studies have confirmed the role of this crayfish in the maintenance of several foodborne pathogenic bacteria. Therefore, the aim of this study was to analyze the main zoonotic bacterial and parasitic pathogens present in a P. clarkii population introduced to the island of Tenerife, Canary Islands, and to assess the potential risk to public health and native fauna. A total of 22 crayfish from Tenerife were analyzed using Biofire FilmArray Gastrointestinal Panels and culture–PCR methods. The results show the presence of Plesiomonas shigelloides, Shigella/enteroinvasive Escherichia coli, enteropathogenic Escherichia coli, Salmonella ser. Enteritidis, Salmonella ser. Typhimurium, and Salmonella ser. Typhi. These results demonstrate the presence of a variety of pathogenic bacteria in the red swamp crayfish in Tenerife that represent a significant concern in terms of public health and conservation. Implementing educational campaigns to inform the community about the risks associated with handling and consuming contaminated crayfish, as well as initiatives for the restoration of the contaminated ecosystem, are necessary to prevent the transmission of the foodborne pathogens.

Pathogen-encoded Rum DNA polymerase drives rapid bacterial drug resistance.

The acquisition of multidrug resistance by pathogenic bacteria is a potentially incipient pandemic. Horizontal transfer of DNA from mobile integrative conjugative elements (ICEs) provides an important way to introduce genes that confer antibiotic (Ab)-resistance in recipient cells. Sizable numbers of SXT/R391 ICEs encode a hypermutagenic Rum DNA polymerase (Rum pol), which has significant homology with Escherichia coli pol V. Here, we show that even under tight transcriptional and post-transcriptional regulation imposed by host bacteria and the R391 ICE itself, Rum pol rapidly accelerates development of multidrug resistance (CIPR, RifR, AmpR) in E. coli in response to SOS-inducing Ab and non-Ab external stressors bleomycin (BLM), ciprofloxacin (CIP) and UV radiation. The impact of Rum pol on the rate of acquisition of drug resistance appears to surpass potential contributions from other cellular processes. We have shown that RecA protein plays a central role in controlling the ability of Rum pol to accelerate antibiotic resistance. A single amino acid substitution in RecA, M197D, acts as a 'Master Regulator' that effectively eliminates the Rum pol-induced Ab resistance. We suggest that Rum pol should be considered as one of the major factors driving development of de novo Ab resistance in pathogens carrying SXT/R391 ICEs.

Design, Synthesis and Bioactivity Evaluation of Ag(I)-, Au(I)- and Au(III)-Quinoxaline-Wingtip N-Heterocyclic Carbene Complexes Against Antibiotic Resistant Bacterial Pathogens.

Intending to homogenize the biological activities of both quinoxaline and imidazole moieties, the proligand, 1-methyl-3-quinoxaline-imidazolium hexaflurophosphate (1.HPF6), and [Ag(1)2][PF6], (2); [Au(1)2][PF6], (3); and [Au(1)Cl3], (4) NHC complexes were synthesized. All the synthesized compounds were characterized by elemental analysis, NMR, and UV-Vis spectroscopy. Finally, single crystal X-ray structures revealed a linear geometry for complex 2 whereas a square planar geometry for complex 4. The formation of complex 3 was confirmed and supported by its MS spectra. The antibacterial activities of all the synthesized complexes were investigated against gram-positive bacteria and gram-negative bacteria. The Au(III)-NHC complex, 4 showed the highest antibacterial activity with extremely low MIC values against both the bacterial strains (0.24 μg mL-1). Monitoring of zeta potential supports the higher activity of complex 4 compared to 2 and 3. ROS production by complex 4 has also been measured in vitro in the CT26 cancer cell lines, which is directly responsible for targetting and killing the bacterial pathogens. Cell cytotoxicity assay using 293T cell lines has been performed to investigate the biocompatibility nature of complex 4. Also, an excellent hemocompatibility was assigned to it from its hemolytic studies, which provide valuable insights into the design of novel antibacterial agents.

Advancements in Bacteriophages for the Fire Blight Pathogen Erwinia amylovora.

Erwinia amylovora, the causative agent of fire blight, causes significant economic losses for farmers worldwide by inflicting severe damage to the production and quality of plants in the Rosaceae family. Historically, fire blight control has primarily relied on the application of copper compounds and antibiotics, such as streptomycin. However, the emergence of antibiotic-resistant strains and growing environmental concerns have highlighted the need for alternative control methods. Recently, there has been a growing interest in adopting bacteriophages (phages) as a biological control strategy. Phages have demonstrated efficacy against the bacterial plant pathogen E. amylovora, including strains that have developed antibiotic resistance. The advantages of phage therapy includes its minimal impact on microbial community equilibrium, the lack of a detrimental impact on plants and beneficial microorganisms, and its capacity to eradicate drug-resistant bacteria. This review addresses recent advances in the isolation and characterization of E. amylovora phages, including their morphology, host range, lysis exertion, genomic characterization, and lysis mechanisms. Furthermore, this review evaluates the environmental tolerance of E. amylovora phages. Despite their potential, E. amylovora phages face certain challenges in practical applications, including stability issues and the risk of lysogenic conversion. This comprehensive review examines the latest developments in the application of phages for controlling fire blight and highlights the potential of E. amylovora phages in plant protection strategies.

Engineering Cyborg Pathogens through Intracellular Hydrogelation.

Synthetic biology primarily focuses on two kinds of cell chassis: living cells and nonliving systems. Living cells are autoreplicating systems that have active metabolism. Nonliving systems, including artificial cells and nanoparticles, are nonreplicating systems typically lacking active metabolism. In recent work, Cyborg bacteria that are nonreplicating-but-metabolically active have been engineered through intracellular hydrogelation. Intracellular hydrogelation is conducted by infusing gel monomers and photoactivators into cells, followed by the activation of polymerization of the gel monomers inside the cells. However, the previous work investigated only Escherichia coli cells. Extending the Cyborg-Cell method to pathogenic bacteria could enable the exploitation of their pathogenic properties in biomedical applications. Here, we focus on different strains of Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae. To synthesize the Cyborg pathogens, we first reveal the impact of different hydrogel concentrations on the metabolism, replication, and intracellular gelation of Cyborg pathogens. Next, we demonstrate that the Cyborg pathogens are taken up by macrophages in a similar magnitude as wild-type pathogens through confocal microscopy and real-time PCR. Finally, we show that the macrophage that takes up the Cyborg pathogen exhibits a similar phenotypic response to the wild-type pathogen. Our work generalizes the intracellular hydrogelation approach from lab strains of E. coli to bacterial pathogens. The new Cyborg pathogens could be applied in biomedical applications ranging from drug delivery to immunotherapy.

Growth stage-related capsular polysaccharide translocon Wza in Vibrio splendidus modifies phage vB_VspM_VS2 susceptibility

Bacteria at different growth stages usually coordinate capsular polysaccharide (CPS) formation and may affect their susceptibility to phage. In this study, we evaluated the infection efficacy of phage vB_VspM_VS2 in V. splendidus AJ01 at different growth stages and explored the role of growth stage-related CPS translocon Wza in the susceptibility of V. splendidus to phage vB_VspM_VS2. The results showed that V. splendidus locked in the stationary growth stage (SGS) or early exponential stage (EES) infected with phage (EES-P) has a low susceptibility to phage vB_VspM_VS and exhibit a pronounced reduction in phage adsorption rate as compared to the EES bacteria. The expression of wza of CPS transport gene was significantly increased in EES bacteria compared to that bacteria locked in the SGS or EES-P. Bacteria with deleted wza (Δwza mutant) escaped phage adsorption due to absence of Wza mediated down-regulation of CPS expression, otherwise. Our results reveal that the Wza of V. splendidus can promotes phage to infect these bacteria via increasing the phage absorption, which provides important implications for using phages therapeutically target pathogenic bacteria in dynamics communities.

Characteristics of Gut Microbiome in the Murine Model of Pancreatic Cancer with Damp-Heat Syndrome.

Murine models of pancreatic cancer with damp-heat syndrome were established based on two methods to explore the differences in the composition of intestinal flora and to seek characteristic genera with potential for model evaluation. In our study, thirty-four C57BL/6J male mice were randomly divided into a control group (Con), a model group (Mod), a classic damp-heat syndrome group (CDHS), and a climate-chamber group (CC). CDHS and CC groups were fed with a high-fat diet and glucose water, while the CDHS group was given 2.4 g/kg alcohol by gavage for 10 days, and the CC group was placed in a climatic chamber with a set temperature of (32 ± 1) °C and humidity of (92 ± 2)% for 10 days. The Mod group, CDHS group, and CC group underwent tumor-building experiments on day 11. Tumorigenicity was then assessed twice a week. After 4 weeks, feces, colon tissue, and tumor tissue were taken from the mice and were tested, and the mice were euthanized afterwards. Mice in the CDHS and CC groups showed symptoms similar to the clinical damp-heat syndrome observed in traditional Chinese medicine (TCM), and exhibited a worse general condition and more rapid tumor growth trend than those in the Mod group. The pathological examination indicated that inflammation was prevalent in the CDHS and CC groups. Both groups had a disrupted intestinal barrier and an overgrowth of pathogenic bacteria such as c_Gammaproteobacteria, o_Enterobacteriales, and g_Bacteroides. Their microbiota composition showed greater diversity. Intestinal flora may have a promising future in the discovery of indicators for evaluating a model of damp-heat syndrome in pancreatic cancer.

Comparative Effects of Crude Extracts and Bioactive Compounds from Bidens pilosa and Bidens alba on Nonspecific Immune Responses and Antibacterial Activity Against Vibrio sp. in Coculture with Lactic Acid Bacteria in Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

This study investigated the effect of substances on nonspecific immune responses of head kidney leukocytes, the antimicrobial activity against Vibrio sp., as well as the time-kill of Vibrio sp. by combining the substances with lactic acid bacteria (LAB) and Pediococcus sp. The substances are B. pilosa hot water extract, B. pilosa powder extract, B. pilosa methanol extract, B. pilosa ethanol extract, B. alba hot water extract, B. alba powder extract, B. alba methanol extract, B. alba ethanol extract, and bioactive compounds, namely cytopiloyne, flavonoid, phenol, ethyl caffeate, luteolin, chlorogenic acid, butein, and linoleic acid. The results showed that some of them were nontoxic to the head kidney leukocytes, which can increase the phagocytic rate, phagocytic index, and respiratory burst. These substances were able to inhibit the growth of Vibrio sp.; they can even completely kill the pathogenic bacteria. The largest of the inhibition zone formed from the EC group at a concentration range of 5-50 µg/mL against V. parahaemolyticus, V. alginolyticus, and V. harveyi with a value of 19.7 ± 0.56, 19.3 ± 1.53, and 20.6 ± 1.53 mm. Furthermore, the time-kill studies showed that the LAB and P. acidilactici can completely kill the Vibrio sp. at 6 h incubation time, mainly in the group of combination with EC.

The pathogen-induced peptide CEP14 is perceived by the receptor-like kinase CEPR2 to promote systemic disease resistance in Arabidopsis.

Secreted plant peptides that trigger cellular signaling are crucial for plant growth, development, and adaptive responses to environmental stresses. In Arabidopsis (Arabidopsis thaliana), the C-TERMINALLY ENCODED PEPTIDE (CEP) family is a class of secreted signaling peptides that is phylogenetically divided into two groups: group I (CEP1-CEP12) and group II (CEP13-CEP15). Several group I CEP peptides regulate root architecture and nitrogen starvation responses, whereas the biological activity and roles of group II CEPs remain unknown. Here, we report that a group II CEP peptide, CEP14, functions as a pathogen-induced elicitor of Arabidopsis immunity. In response to infection by the bacterial pathogen Pseudomonas syringae, CEP14 expression was highly induced via the salicylic acid pathway in Arabidopsis leaves and roots. In the absence of pathogen attack, treatment of Arabidopsis plants with synthetic CEP14 peptides was sufficient to trigger immune responses. Genetic and biochemical analyses demonstrated that the receptor-like kinase CEP RECEPTOR 2 (CEPR2) perceives CEP14 to trigger plant immunity. The SOMATIC EMBRYOGENESIS RECEPTOR KINASES (SERKs) BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) and SERK4 also participated in CEP14 perception by forming CEP14-induced complexes with CEPR2. Overexpression of CEP14 largely enhanced Arabidopsis resistance to P. syringae, while CEP14 or CEPR2 mutation significantly attenuated Arabidopsis systemic resistance to P. syringae. Taken together, our data reveal that the pathogen-induced CEP14 peptide, which is perceived by the CEPR2-BAK1/SERK4 receptor complexes, acts as an endogenous elicitor to promote systemic disease resistance in Arabidopsis.

Features of the microbiome of burial soils

Introduction. On the territory of the Russian Federation, as well as worldwide, a large amount of space is allocated for burials. The soils found in the burial areas have their own characteristics. In the scientific literature there is a few works devoted to the problem of hygienic assessment of cemeteries from the point of view of their impact on the environment, as well as on the population living next to necrosols or working on them. Depending on the chemical and biological effects, the microbial composition of the soil changes. This process is influenced by many factors, including humidity, the initial content of organic and mineral substances, level of acidity, structure of the soil and peculiarities of the course of intra-soil gas-phase reactions. The key participants in the decomposition of organic material are bacteria and fungi, the diversity and dynamics of which directly depend on the degree of soil contamination with pollutants. The purpose of this study was to analyze the microbiota of cemetery soil in its various layers. Materials and methods. The cemeteries of Moscow (NikoloKhovanskoye, Nikolo-Arkhangelskoye, Perepechenskoye), Moscow (Mytishchenskoye, Domodedovo), Tula (Municipal Cemetery No. 1 of the Municipal State Enterprise of the Municipal Formation of the City of Tula Combine of Specialized Services), Kursk (old city cemetery Kurchatov), Krasnoyarsk (Zheleznogorsk) regions and Altai (Yarovoye) Krais were selected as research objects for assessing the microbiome of soils. Results. The most common bacterial pathogens were found to be Enterococcus spp. (81%), Bacillus spp. (75%) and E. coli (45.1%). Mushrooms of Penicillium spp.. were isolated from 61% of the samples. The revealed microbiota profiles of the samples of the studied cemetery soils reflect the microbial composition of humans, which allows substantiating the main methods and algorithm for identifying decomposition processes depending on the time frame of burials. Limitations. The limitation of the study is due to the risk of contact with cemetery soil was not assessed, since not all genera of isolated microorganisms could be identified by their biochemical properties. Conclusion. Studies conducted in cemetery areas have shown how diverse the soil microbiome is in burial sites and varies depending on the depth of sampling. The microbiota profiles of cemetery soil samples identified during the study reflect the lifetime microbial composition of the human body, which makes it possible to substantiate approaches to identifying decomposition processes depending on the time frame of burials.

Overexpression of l,d-Transpeptidase A Induces Dispensability of Rod Complex in Escherichia coli.

Antimicrobial resistance (AMR) is a significant global threat, and the presence of resistance-determinant genes is one of the major driving forces behind it. The bacterial rod complex is an essential set of proteins that is crucial for cell survival due to its role in cell wall biogenesis and shape maintenance. Therefore, these proteins offer excellent potential as drug targets; however, compensatory mutations in nontarget genes render this complex nonessential. The MreB protein of this complex is an actin homologue that rotates along the longitudinal axis of the cell to provide rod shape to the bacteria. In this study, using chemical-chemical interaction profiling and FtsZ suppression assay, we identified the MreB targeting activity of IITR07865, a previously discovered small molecule in our lab. Escherichia coli suppressors against IITR07865 revealed mutations in two cell division-associated genes, min C and pal, that have not been previously implicated in rod complex essentiality. IITR07865 resistant mutants were found to inactivate and render the rod complex nonessential, making the rod complex inhibitors ineffective. Further, through transcriptome analysis, we reveal the primary cause of resistance in suppressor strains to be the overexpression of an l, d-transpeptidase A enzyme, which is involved in peptidoglycan and Braun's lipoprotein cross-linking. Our results demonstrate a novel mechanism of resistance development in rod-shaped Gram-negative bacterial pathogen E. coli involved in UTIs where mecillinam, a clinically used antibiotic that targets rod complex, is a drug of choice.

Selective colonization of microplastics, wood and glass by antimicrobial-resistant and pathogenic bacteria.

The Plastisphere is a novel niche whereby microbial communities attach to plastic debris, including microplastics. These communities can be distinct from those found in the surrounding environment or those attached to natural substrates and may serve as a reservoir of both pathogenic and antimicrobial-resistant (AMR) bacteria. Owing to the frequent omission of appropriate comparator particles (e.g. natural substrates) in previous studies, there is a lack of empirical evidence supporting the unique risks posed by microplastics in terms of enrichment and spread of AMR pathogens. This study investigated selective colonization by a sewage community on environmentally sampled microplastics with three different polymers, sources and morphologies, alongside natural substrate (wood), inert substrate (glass) and free-living/planktonic community controls. Culture and molecular methods (quantitative polymerase chain reaction (qPCR)) were used to ascertain phenotypic and genotypic AMR prevalence, respectively, and multiplex colony PCR was used to identify extra-intestinal pathogenic Escherichia coli (ExPECs). From this, polystyrene and wood particles were found to significantly enrich AMR bacteria, whereas sewage-sourced bio-beads significantly enriched ExPECs. Polystyrene and wood were the least smooth particles, and so the importance of particle roughness on AMR prevalence was then directly investigated by comparing the colonization of virgin vs artificially weathered polyethylene particles. Surface weathering did not have a significant effect on the AMR prevalence of colonized particles. Our results suggest that the colonization of plastic and non-plastic particles by AMR and pathogenic bacteria may be enhanced by substrate-specific traits.

Inflammasome-mediated pyroptosis in defense against pathogenic bacteria.

Macrophages, neutrophils, and epithelial cells are pivotal components of the host's immune response against bacterial infections. These cells employ inflammasomes to detect various microbial stimuli during infection, triggering an inflammatory response aimed at eradicating the pathogens. Among these inflammatory responses, pyroptosis, a lytic form of cell death, plays a crucial role in eliminating replicating bacteria and recruiting immune cells to combat the invading pathogen. The immunological function of pyroptosis varies across macrophages, neutrophils, and epithelial cells, aligning with their specific roles within the innate immune system. This review centers on elucidating the role of pyroptosis in resisting gram-negative bacterial infections, with a particular focus on the mechanisms at play in macrophages, neutrophils, and intestinal epithelial cells. Additionally, we underscore the cell type-specific roles of pyroptosis invivo in these contexts during defense.