Microbiology Research

Rhizosphere microorganisms effectively exploit nutrient resources within the rhizosphere, while growth-promoting bacteria in this environment play a vital role in regulating soil fertility and enhancing plant health. In this study, we utilized a comprehensive approach that included the isolation, purification, and identification of dominant microorganisms, alongside high-throughput sequencing technology. This methodology was employed to analyze the primary microbial groups and their diversity within the rhizosphere soil of Helichrysum arenarium (L.) Moench in Altay, Xinjiang, China. By isolating bacterial strains from the rhizosphere soil using a dilution coating method, we successfully obtained 43 distinct strains. Subsequently, selective media were employed to screen for growth-promoting characteristics among these isolated strains derived from the rhizosphere soil of H. arenarium (L.) Moench. The results, obtained through high-throughput amplification sequencing, revealed diverse bacterial communities belonging to 35 phyla, 93 orders, 215 families, 324 genera, and 231 species associated with H. arenarium (L.) Moench, as well as fungal communities comprising 14 phyla, 47 orders, 96 families, 204 genera, and 571 species present in the rhizosphere soil. Among these identified communities, Actinobacteriota emerged as the predominant bacterial phylum while Ascomycetes and Mortieromycetes were recognized as the principal fungal phyla found in the rhizospheric soil of H. arenarium (L.) Moench. Analysis of culturable bacteria’s promotion activity within this rhizospheric environment indicated that three strains—S16, S31, and S29—exhibited the highest solubility index for inorganic phosphorus; additionally, the screened strains S7 and S10 demonstrated nitrogen-fixing capabilities. Furthermore, ten strains exhibiting excellent iron-bearing capacities were identified; notably, strain S16 displayed the highest D/d value indicating, its superior iron-bearing capacity. The growth-promoting bacteria were identified as Kocuria rosea, Priestia megaterium, Bacillus mobilis, Bacillus bataviensis, three variants of Bacillus mycoides, Bacillus paramobilis, Bacillus sonorensis, and Alcaligenes faecalis. This study provides a foundational understanding of how microorganisms in the rhizosphere of H. arenarium (L.) Moench influence soil nutrient release and offers valuable insights into enhancing yield and quality cultivation by isolating, screening, and identifying growth-promoting bacteria from rhizosphere soil.

An EBSL-encoding plasmid, pESBL-PH, was identified during a nosocomial outbreak of Klebsiella pneumoniae ST628 at a United Kingdom general district hospital in 2018. The plasmid from the earliest 2018 K. pneumoniae strain discovered during the outbreak was assembled using both Oxford nanopore long reads and illumina short reads, yielding a fully closed plasmid, pESBL-PH-2018. pESBL-PH-2018 was queried against the complete NCBI RefSeq Plasmid Database, comprising 93,823 plasmids, which was downloaded on 16 July 2024. To identify structurally similar plasmids, strict thresholds were applied, including a mash similarity ≥0.98. This returned 61 plasmids belonging to 13 unique sequence types (STs) hosts. The plasmids were detected from 13 unique countries, dating from 2012 to 2023. The AMR region of the plasmids varied. Interestingly IS26-mediated tandem amplification of resistance genes, including the ESBL gene blaCTX-M-15 was identified in two independent strains, raising their copy number to three. Furthermore, the genomic background of strains carrying a pESBL-PH-2018-like plasmid were analyzed, revealing truncation of the chromosomal ompK36 porin gene and carbapenem resistance gene carriage on accessory plasmids in 17.85% and 26.78% of strains with a complete chromosome available. This analysis reveals the widespread dissemination of an ESBL-encoding plasmid in a background of resistance-encoding strains, requiring active surveillance.

Rapid environmental changes driven by human activities are contributing to a significant decline in global biodiversity, with avian species being particularly affected due to their migratory behavior. As highly mobile hosts, birds facilitate the geographic dispersal of ectoparasites, including ticks, which serve as vectors for numerous zoonotic pathogens. This study, conducted in collaboration with the Faunistic Observatory of the Asinara National Park between 2021 and 2023, aimed to investigate the potential role of migratory birds in tick dispersal and the presence of Borrelia spp. DNA. Birds were captured using mist nets during pre-breeding (April–May) and post-breeding (October–November) migration periods. Ticks were systematically collected and identified at the species level, and molecular analyses were performed using real-time and conventional PCR to detect the presence of Borrelia spp. DNA. Results showed a distinct seasonal variation in tick species composition. In autumn, Ixodes ricinus was predominant (99%), whereas Hyalomma species were more frequently observed in spring (78%). Molecular screening revealed Borrelia spp. DNA in 26.1% of the collected ticks, with Borrelia garinii being the most prevalent species. These findings underscore the ecological significance of migratory birds in the dissemination of ticks and tick-borne pathogens, highlighting their potential role in shaping disease transmission dynamics across different geographic regions. This study provides valuable insights into the seasonal fluctuations in tick populations associated with migratory avifauna and the epidemiological risks posed by these interactions. Continued surveillance of migratory birds as vectors of zoonotic pathogens is essential for informing public health strategies and mitigating the risks of emerging infectious diseases, but further investigation is needed to clarify the actual role of migratory birds in the transmission of Borrelia spp.

Vibrio parahaemolyticus is a foodborne pathogen commonly associated with the consumption of contaminated seafood, particularly oysters. While PCR and real-time PCR are widely used to detect its pathogenicity through tdh and trh gene detection, these methods may not be practical in resource-limited settings such as field environments. To address this limitation, a rapid, sensitive, and specific duplex detection method was developed using the multienzyme isothermal rapid amplification (MIRA) assay in combination with lateral flow dipstick (LFD) technology. The assay utilized specific primer sets and probes to simultaneously amplify tdh and trh fragments tagged with 3′-FAM and 5′-Digoxigenin or Biotin during MIRA amplification, enabling the detection via respective antibody capture on the LFD strip. This duplex MIRA-LFD assay demonstrated a detection limit of 100 fg of DNA, 300 CFU/reaction for bacterial culture, and 3000 CFU/reaction for seeded oyster samples at 40 °C within 20 min. Notably, the assay exhibited no cross-reactivity with nine other Vibrio species or 18 foodborne pathogens, confirming its high specificity. Due to its simplicity, rapid turnaround time, and high sensitivity, this duplex MIRA-LFD assay offers a valuable tool for the surveillance of V. parahaemolyticus pathogenicity, aiding in public health protection and supporting the local seafood industry.

The Clonostachys genus is a saprophytic soil microfungus (Ascomycota). It exhibits significant ecological adaptability and plays a crucial role in maintaining the balance of soil microorganisms. Species within this genus are natural antagonists of insects and nematodes, and they also combat phytopathogenic fungi through mycoparasitism. This process involves producing lytic enzymes and competing for space and nutrients. Clonostachys species are effective biocontrol agents in agriculture and have been utilized to manage pests affecting many high-value commercial crops, acting as a natural biopesticide. They inhabit plant tissues, boosting plant defenses and activating genes for water and nutrient uptake, enhancing plant performance. Additionally, they produce enzymes and bioactive metabolites with antimicrobial, antifungal, nematocidal, anticancer, and antioxidant properties. Clonostachys species can degrade plastic waste and remove hydrocarbons from crude oil-contaminated sites when functioning as endophytes, positioning Clonostachys as a promising candidate for reducing environmental pollution. There are still challenges and limitations, such as the continuous surveillance of the safety of Clonostachys species on plants, the establishment of commercial applications, formulation viability, and variability due to field conditions. These issues will have to be addressed. This review provides an overview of Clonostachys ecology, morphology, classification, and biotechnological applications, emphasizing its significance in various fields.

Canine monocytic ehrlichiosis (CME) is a highly infectious disease with zoonotic potential. Ehrlichia canis, the causative agent, is primarily transmitted by Rhipicephalus sanguineus. Tick infestation and tick-borne diseases (TBDs) are serious human and veterinary health problems due to the lack of efficient control measures. This review aims to provide information about CME, detailing epidemiology, pathogenesis, clinical manifestations, and current strategies for diagnosis, treatment, and prevention. The review delves into the biology of R. sanguineus, exploring its lifecycle, habitat, and mechanisms of E. canis transmission. Ehrlichia canis infection follows a three-phase transition: acute, subclinical, and chronic, with distinct clinical signs, from mild to severe and even life-threatening in some cases, with the potential for asymptomatic long-term carriers acting as reservoirs. Pathogenesis involves complex interactions between the pathogen and host immune responses, influencing disease severity and progression. Additionally, the review highlights challenges in controlling CME due to the widespread distribution of R. sanguineus. Genetic diversity within R. sanguineus populations and their varying vector competence further complicate control efforts. The role of environmental changes in tick distribution aligns the control of CME within the One Health concept, integrating approaches to improve outcomes for humans, animals, and the environment. Preventive measures on tick control and potential vaccine development are also reviewed.

The aim of our study was to evaluate the discriminative value of gene and protein expression levels of the inflammatory marker (YKL-40) and lysosome-associated membrane protein 1 and 2 (LAMP-1 and LAMP-2) in patients with central nervous system (CNS) infections. Thirty hospitalized patients with CNS infections and undefined etiology, and 22 healthy subjects as a control group, were included in the study. Gene expression levels of YKL-40, LAMP-1 and LAMP-2 were determined by qPCR. Plasma and CSF concentrations of the tree proteins of interest were detected by ELISA. Our results showed that mRNA levels of YKL-40 were significantly downregulated in WBCs of patients compared to controls, while plasma YKL-40 concentrations were higher. LAMP-1 significantly increased in patients’ plasma and CSF was found. Patients were subdivided depending on the confirmed or presumed etiological agent into two subgroups groups—patients with bacterial or with viral neuroinfection. Differences between plasma levels of YKL-40 in the subgroups when matched with controls were detected. The concentrations of the glycoprotein were higher in patients with bacterial infections compared to those with the viral ones. We revealed that LAMP-1 plasma levels were also significantly increased in patients with viral infections in comparison to healthy individuals. We could speculate that YKL-40 plasma levels might serve as a fast discriminative tool to support the presence of viral or bacterial CNS infections.

Antimicrobial resistance is increasingly becoming a serious public health issue. There is scientific evidence linking the use of antibiotics in livestock production to the emergence and spread of resistance in bacteria that are important for human health. To assess the prevalence of antimicrobial resistance in Escherichia coli and Enterococcus spp., fecal and slurry wastewater samples were collected from various cattle and swine farms, mainly located in the northern and central regions of Portugal. Samples from each farm were pooled for microbiological processing to isolate Escherichia coli and Enterococcus spp., followed by specific antibiotic susceptibility testing for each species using the disk diffusion method. The results of these analyses indicated a significant issue with tetracycline resistance in E. coli and Enterococcus spp. Furthermore, a notably higher frequency in resistant strains was observed in the majority of slurry samples compared to those derived from swine feces. This observation led to the hypothesis that slurry may provide a comprehensive historical perspective for studying the antibiotic resistance patterns present on a farm.

West Nile Virus (WNV), a mosquito-borne pathogen, is a growing public health challenge across Europe. Environmental and anthropogenic factors have led to the spread of the virus to higher geographic latitudes, as well as to increased viral circulation and genetic diversity. Aims: This study aimed to describe the epidemiological, clinical, and laboratory characteristics of WNV cases in Bulgaria during 2024 and to investigate WNV phylogenetics. Epidemiological, clinical and laboratory data from 32 patients with confirmed or probable WNV infections were collected and analysed. Complete viral genomes were obtained from two samples using whole genome sequencing (WGS). Phylogenetic analysis was performed using the Nextstrain WNV analysis pipeline. Severe disease was observed in 21 patients, with three fatalities reported in older males with comorbidities. Phylogenetic analysis revealed that Bulgarian strains clustered within the Central/Southern European clade of lineage 2, closely related to Greek strains. Evidence suggested localised viral evolution following cross-border introduction from Greece. Our study provides a detailed clinical and laboratory characterization of the human WNV cases detected in Bulgaria in 2024. Improved diagnostic workflows, expanded laboratory resources and increased molecular surveillance are essential to better understand the burden of WNV infections in Bulgaria, as well as to follow the evolution and spread of the virus.

Carbapenemase-producing Klebsiella pneumoniae is responsible for multiple serious infections with high mortality rates. K. pneumoniae carbapenemases (KPCs) are the most commonly isolated carbapenemases worldwide. To study the epidemiological and molecular characteristics of KPC-producing K. pneumoniae (KPC-KP), we conducted a retrospective study at the University General Hospital of Ioannina, Greece. A total of 177 K. pneumoniae clinical strains from the period 2014–2015 were confirmed as KPC producers by polymerase chain reaction (PCR) and were further examined for the presence of blaVIM, blaNDM, blaTEM, blaSHV, and blaCTX-M genes. Using the amplification refractory mutation system (ARMS) method, we identified the presence of the KPC-2 allele in 130 strains and the KPC-9 allele in 47. Strains from both allele groups belonged to the sequence type 258 (ST258). KPC-9 was responsible for a distinct outbreak, considered part of the broader KPC-2 outbreak. Molecular characterization of selected KPC-KP isolates from the period 2021–2022 revealed their continued presence in our hospital. Comparison of the antimicrobial susceptibility profiles of the two alleles showed a statistically significant increase in minimum inhibitory concentration (MIC) for ceftazidime (p = 0.03) and higher resistance to amikacin (p = 0.012) and colistin (p < 0.001) for KPC-9 compared to the KPC-2 allele. The two KPC alleles had similar mortality rates. This study demonstrates the heterogeneity of resistance genes in carbapenem-resistant K. pneumoniae (CR-KP) within a single-hospital setting and underscores the need for immediate containment measures.

The United States is facing outbreaks of highly pathogenic avian influenza H5N1 in birds and dairy cattle, along with threats of African swine fever, classical swine fever, and foot-and-mouth disease. While the National Animal Health Laboratory Network (NAHLN) depends on high-throughput testing, the KingFisher Duo Prime, IndiMag 48s, and IndiMag 2 are viable alternatives to aid in outbreak assessments. This study evaluates extraction platforms and the IndiMag Pathogen Kit for detecting the previous listed pathogens. Samples and reference materials were extracted using the MagMAX Viral RNA Isolation Kit, MagMAX CORE Nucleic Acid Purification Kit, and IndiMag Pathogen Kit. Real-time RT-PCR was performed following NAHLN protocols to assess analytical and diagnostic performance. Comparable limits of detection across extraction chemistries, instrumentation, and pathogens were demonstrated, with PCR efficiency ranging between 82.5% and 107.6%. The precision variability was low, with the coefficient of variation ranging from 0.16% to 1.76%. Diagnostic sensitivity and specificity were 100%, with a kappa coefficient of 1.0, indicating strong agreement between methods. These findings support the KingFisher Duo Prime, IndiMag 48s, IndiMag 2, and IndiMag Pathogen Kits as reliable options for NAHLN-approved testing, increasing equipment and reagent alternatives to enhance diagnostic resilience and improve response capabilities to emerging animal health threats.

Introduction: Antimicrobial resistance (AMR) is a critical global public health issue, leading to prolonged illness, increased morbidity and mortality, and rising healthcare costs. The effectiveness of antibiotics is diminishing due to the emergence of resistant bacterial strains. This study aimed to determine the bacterial profile and AMR patterns of clinical isolates at Uttara Adhunik Medical College Hospital (UAMCH), Dhaka. Methods: A retrospective study was conducted at UAMCH from January 2017 to December 2019. A total of 32,187 clinical specimens (urine, blood, stool, wound swabs/pus, and sputum) were processed, of which 4232 yielded positive cultures. Bacterial identification followed standard bacteriological methods, and antibiotic susceptibility was assessed using the Kirby–Bauer disc diffusion method per CLSI guidelines. Data analysis was conducted using WHONET and QAAPT. Results: The highest proportion of positive cultures was from urine (47.5%), followed by blood (35.0%) and wound swabs/pus (10.1%). The most common isolates were Escherichia coli (37.2%), Salmonella typhi (25.7%), and Klebsiella sp. (11.5%). Gram-negative bacteria showed high resistance to commonly used antibiotics such as amoxicillin/clavulanic acid, cefixime, and ceftriaxone, while the resistance rates were lower for gentamicin, amikacin, and meropenem. However, Acinetobacter sp. exhibited alarming resistance to all tested antibiotics. Conclusions: This study highlights concerning resistance patterns among bacterial isolates, emphasizing the need for ongoing AMR surveillance to inform treatment strategies and improve patient care in Bangladesh.

Foodborne diseases (FBDs) represent a significant public health concern, particularly in regions like Central America and the Caribbean (CAC), where surveillance gaps due to a lack of resources, knowledge, and technical abilities hinder control over outbreaks. This review investigates the bacterial pathogens responsible for FBDs, their prevalence, management challenges, and prevention strategies. This systematic review followed PRISMA guidelines, focusing on bacterial FBDs in CAC from 2000 to 2024. PubMed and Google Scholar were used as primary databases, supported by other sources to identify relevant studies. Inclusion criteria encompassed studies focusing on bacterial pathogens, prevalence, risk factors, and surveillance practices. Out of the 509 studies initially identified, 35 met the inclusion criteria. The most prevalent pathogens were Salmonella spp., Escherichia coli, Campylobacter spp., and Aliarcobacter spp., with contamination often associated with poultry, eggs, and vegetables. Key challenges included inadequate surveillance systems, limited resources, and inconsistent reporting practices. A more significant investment in pathogen monitoring, documentation, and education, along with technologies like whole-genome sequencing (WGS), is crucial. Institutional and governmental funding is vital to improve surveillance and strengthen regional risk analysis.

Infections from the protozoa Entamoeba histolytica (E. histolytica), Giardia lamblia (G. lamblia), and Trichomonas vaginalis (T. vaginalis) pose a public health issue, with albendazole and mebendazole serving as the second-line medications for treating these parasitic infections. However, the low aqueous solubility of these compounds has led to the exploration of new strategies to enhance their solubility, with the formation of salts being a commonly employed strategy. The sulfonates A1, A2, and A3 of albendazole, along with M1, M2, and M3 of mebendazole, were synthesized. The antiparasitic activity in vitro was assessed against the trophozoites of E. histolytica, G. lamblia, and T. vaginalis. The salts A2, A3, M2, and M3 demonstrated a greater antiparasitic effect (IC50 37.95–125.53 µM) compared to the positive controls albendazole and mebendazole. The salts A1, A3, M2, and M3 do not exhibit cytotoxic effects at concentrations of 500 µM on the Vero cell line. Taken together, these findings indicate that the formation of these new solid saline phases enhances the antiparasitic effects in vitro, which is crucial in the current search for improved, safe, and effective antiparasitic agents.

Staphylococcus aureus (S. aureus) is a disreputable symbiotic bacterium that is responsible for a range of diseases, including life-threatening pneumonia, endocarditis, septicemia, and others, which has led to an immense loss in both public health and economy, imposing a significant burden on society. To investigate the pathogenic mechanism of S. aureus and develop new treatment methods for infectious diseases caused by S. aureus, various rabbit models have been developed to simulate different infections by S. aureus, such as pneumonia models, meningitis models, and endocarditis models, etc. In this review, we summarized the application of rabbit models in S. aureus-induced infectious diseases.

Cocoa fermentation is a critical step in chocolate production, influencing the flavor, aroma, and overall quality of the final product. This review focuses on the microbial dynamics of cocoa fermentation, emphasizing the roles of yeasts, lactic acid bacteria (LAB), and acetic acid bacteria (AAB). These microorganisms interact in a well-defined succession, producing organoleptic compounds such as alcohols, organic acids, and esters, which are key to the sensory profile of cocoa. This article examines the impact of different fermentation methods, including spontaneous fermentation and the use of starter cultures, on microbial communities and flavor development. Advances in starter culture technology are highlighted, demonstrating how microbial control can enhance fermentation efficiency, reduce fermentation time, and improve the consistency of chocolate flavor. Patents related to cocoa fermentation further emphasize the growing interest in microbial management to meet market demands for high-quality, distinct chocolate. This review also outlines future research directions, including the identification of new microbial strains, optimization of fermentation conditions, and the potential of biotechnological advancements to improve the fermentation process. Understanding microbial dynamics in cocoa fermentation offers significant potential for enhancing chocolate quality, sustainability, and the development of new, region-specific flavor profiles.

The objective of this study was to determine the frequency of Salmonella in pig samples analyzed at the Veterinary Diagnostic Unit of the Faculty of Agricultural Sciences of the University of Antioquia, Colombia, between 2022 and 2023. Out of 5820 serum samples analyzed using indirect enzyme-linked immunosorbent assay, 63.76% were positive. Additionally, Salmonella was isolated and identified in a separate group of 848 samples (feces, tissues, etc.) with a positivity rate of 23.47%. Eight serotypes were identified, with the most common being Salmonella enterica subsp. enterica serotype Typhimurium (49.2%), followed by its monophasic variant I 4,[5],12:i:- (23%), and serotype Choleraesuis (18%). Antimicrobial susceptibility profiles of 105 isolates were evaluated using the Kirby–Bauer method, which demonstrated higher resistance (100%) to ampicillin, tiamulin, penicillin, tylosin, and erythromycin (these were the least tested), followed by florfenicol (44/54), doxycycline (4/5), spectinomycin (18/25), amoxicillin (32/46), chloramphenicol (2/3), tetracycline (2/3), and enrofloxacin (34/64), with lower resistance observed for fosfomycin (2/38) and ceftiofur (5/35). Multi-drug resistance was observed in 59% (62/105) of the isolates. The high proportion of Salmonella and the levels of resistance to various drugs raise significant concerns, indicating potential deficiencies in responsible antimicrobial use and management practices on pig farms in the region.

This study investigated the effect of different culture agar media, derived from Bennett’s medium, on the antimicrobial activity of 15 Streptomyces sp. and 1 Lentzea sp. strains isolated from mining environments. The media were prepared from the standard Bennett’s medium by suppressing one, two, or three ingredients—yeast extract (YE), beef extract (BE), or casein (Cas)—while maintaining glucose (Gluc) or by substituting it with fructose (Fruc) or galactose (Gal) and keeping the same suppressions. The antimicrobial activity was investigated against Candida albicans ATCC 10231, Staphylococcus aureus ATCC 29213, Bacillus subtilis ATCC 6633, and Escherichia coli K12. The antimicrobial activity of actinomycete strains was positively influenced by media modifications, though the response was actinomycete strain and target pathogen-dependent. Unexpectedly, thirteen strains exhibited poor growth on a pure agar-agar medium, including six Streptomyces strains (AS34, AS3, BS59, BS68, BS69, and DAS104) that showed notable antimicrobial activity, with inhibition zone diameters ranging from 10.75 ± 1.06 to 18.00 ± 0.00 mm. Modifications of Bennett’s medium, including replacing glucose with fructose or galactose and maintaining yeast extract or both yeast extract and beef extract, induced and enhanced the antimicrobial activity of several actinomycete strains. Notably, the new media induced antimicrobial activity in strains that showed no activity in Bennett’s medium. They led, compared to Bennett’s medium, to the detection of eight additional active strains against S. aureus, eight against B. subtilis, six against E. coli, and four against C. albicans. This study is the first to explore the modification of Bennett’s medium, either by subtraction or substitution, in order to investigate the effect on antimicrobial activity of actinomycete strains. These results highlight the importance of the composition of culture media on inducing or boosting antimicrobial activity in Streptomyces and Lentzea.

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) are a major healthcare challenge, necessitating effective antimicrobial therapy for treatment. However, the prevalence of antimicrobial resistance among UPEC strains is escalating, particularly among patients experiencing recurrent infection. The rise in UPEC strains that exhibit resistance to multiple antimicrobial agents, including the spread of extended-spectrum beta-lactamase (ESBL)-producing UPEC, intensifies the complexity of managing UTIs. Genetic variations within UPEC strains play a major role in their ability to resist antimicrobial agents and adapt to changing environments. Unveiling and understanding the genomic landscape of emerging UPEC strains is essential for comprehending the genetic basis of their resilience. Moreover, monitoring these genetic strains is crucial for identifying patterns of resistance dissemination, guiding infection control measures, and informing the development of targeted therapeutics.

Soda lakes are extreme saline–alkaline environments that harbor metabolically versatile microbial communities with significant biotechnological potential. This study employed shotgun metagenomics (NovaSeq PE150) to investigate the functional diversity and metabolic potential of microbial communities in Ethiopia’s Chitu and Shala Lakes. An analysis of gene content revealed 554,609 and 525,097 unique genes in Chitu and Shala, respectively, in addition to a substantial fraction (1,253,334 genes) shared between the two, underscoring significant functional overlap. Taxonomic analysis revealed a diverse phylogenetic composition, with bacteria (89% in Chitu Lake, 92% in Shala Lake) and archaea (4% in Chitu Lake, 0.8% in Shala Lake) as the dominant domains, alongside eukaryotes and viruses. Predominant bacterial phyla included Pseudomonadota, Actinomycetota, and Gemmatimonadota, while Euryarchaeota and Nitrososphaerota were prominent among archaea. Key genera identified in both lakes were Nitriliruptor, Halomonas, Wenzhouxiangella, Thioalkalivibrio, Aliidiomarina, Aquisalimonas, and Alkalicoccus. Functional annotation using the KEGG, eggNOG, and CAZy databases revealed that the identified unigenes were associated with various functions. Notably, genes related to amino acid, carbohydrate, and energy metabolism (KEGG levels 1–2) were predominant, indicating that conserved core metabolic functions are essential for microbial survival in extreme conditions. Higher-level pathways included quorum sensing, two-component signal transduction, and ABC transporters (KEGG level 3), facilitating environmental adaptation, stress response, and nutrient acquisition. The eggNOG annotation revealed that 13% of identified genes remain uncharacterized, representing a vast untapped reservoir of novel enzymes and biochemical pathways with potential applications in biofuels, bioremediation, and synthetic biology. This study identified 375 unique metabolic pathways, including those involved in pyruvate metabolism, xenobiotic degradation, lipid metabolism, and oxidative stress resistance, underscoring the microbial communities’ ability to thrive under fluctuating salinity and alkalinity. The presence of carbohydrate-active enzymes (CAZymes), such as glycoside hydrolases, polysaccharide lyases, and oxidoreductases, highlights their role in biomass degradation and carbon cycling. Enzymes such as alkaline proteases (Apr), lipases (Lip), and cellulases further support the lakes’ potential as sources of extremophilic biocatalysts. These findings position soda lakes as reservoirs of microbial innovation for extremophile biotechnology. Future research on unannotated genes and enzyme optimization promises sustainable solutions in bioenergy, agriculture, and environmental management.

The development of microbial-based biostimulants to enhance the growth of crops and support a healthy and sustainable soil requires the isolation and large-scale industrial culture of effective microorganisms. In this study, work was undertaken to isolate and characterize free-living nitrogen-fixing bacteria capable of acting as biostimulants alone or by incorporation into and/or supplementation with a current commercial crop biostimulant for farmers. Free-living bacteria were isolated from soil, sugar cane mulch, and plant roots following preliminary culture in a nitrogen-free media that targeted specific groups of known diazotrophs. Following the identification of each isolate by 16S rDNA sequence analysis, isolates selected for further study were identified as most closely related to Priestia megaterium, Sphingobium yanoikuyae, and Burkholderia paludis. Each isolate was investigated for its capacity to promote plant growth in nitrogen-free media. Wheat seedlings were inoculated with the isolates separately, together as a consortium, or in combination with the commercial biostimulant, Great Land Plus®. Compared to no-treatment control plants, the fresh weights were higher in both the shoots (183.2 mg vs. 330.6 mg; p < 0.05) and roots (320.4 mg vs. 731.3 mg; p < 0.05) of wheat seedlings inoculated with P. megaterium. The fresh weights were also higher in the shoots (267.8 mg; p < 0.05) and roots (610.3 mg; p = 0.05) of wheat seedlings inoculated with S. yanoikuyae. In contrast, the fresh weight of the shoot and root systems of plants inoculated with B. paludis were significantly lower (p < 0.05) than that of the no-treatment control plants. Moreover, when Great Land Plus® was supplemented with a consortium of P. megaterium and S. yanoikuyae, or a consortium of P. megaterium, S. yanoikuyae, and B. paludis no promotion of plant growth was observed.

Multidrug-resistant (MDR) bacterial pathogens pose a serious threat to global health, underscoring the urgent need for innovative therapeutic strategies. In this work, we designed and characterized thiol-modified antisense oligonucleotide-capped gold nanoparticles (ASO-AuNPs) to resensitize antibiotic-resistant bacteria. Transmission electron microscopy and UV–Vis spectroscopy confirmed the morphology, size, and optical properties of AuNPs and ASO-AuNPs. Minimum inhibitory concentrations (MIC) of ampicillin were determined for non-resistant Escherichia coli DH5α (16 ppm) and an ampicillin-resistant E. coli DH5α strain (PSK, 32,768 ppm). When co-administered with ampicillin, ASO-AuNPs (0.1 and 0.2 nM) significantly reduced bacterial growth compared to the antibiotic-alone control (p < 0.05), demonstrating the capacity of ASO-AuNPs to restore antibiotic efficacy. These findings provide a proof of concept that antisense oligonucleotide-functionalized nanomaterials can be harnessed to overcome beta-lactam resistance, setting the stage for further optimization and translation into clinical applications.

Tilapia Parvovirus (TiPV) is a rising pathogen responsible for high mortality in tilapia aquaculture. Understanding TiPV’s pathogenesis is crucial for developing effective management strategies. This study aimed to elucidate TiPV pathogenesis by evaluating its cytotoxic effects on Danio rerio gill (DRG) cell monolayers and its impact on host immune responses. PCR-confirmed TiPV-infected DRG cell monolayers were subjected to an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay at 24, 48, 72, and 96 h post-infection to assess cell viability and cytotoxicity. The MTT assay revealed a progressive decline in DRG cell viability over time, with viable cell percentages decreasing from 66.71% at 24 h to 31.28% at 96 h in TiPV-infected cultures, compared to consistently high viability in controls. Simultaneously, quantitative real-time PCR (qPCR) was used to assess the expression of key immune-related genes, including Interleukins (IL-1β, IL-8), Toll-like receptor 7 (TLR7), Major Histocompatibility Complex II (MHC-II), Tumor Necrosis Factor α (TNF-α), Nuclear Factor Kappa B (NF-κB), and Chemokine Receptors (CRs).qPCR analysis showed an upregulation of IL-8, IL-1β, TNF-α, and CRs, indicating an early inflammatory response. However, significant downregulation of TLR7, MHC-II, and NF-κB suggests TiPV’s ability to modulate host immune responses. The results highlight that TiPV induces significant cytotoxicity in DRG cells, leading to severe cellular damage. The virus also alters host immune responses by modulating the expression of key immune genes, which may contribute to its virulence and persistence. These findings enhance our understanding of TiPV pathogenesis and highlight the need for targeted research to develop effective control strategies for TiPV in aquaculture systems.

Microbial metallophores are low-molecular-weight chelating agents produced by microorganisms to acquire essential metal ions. Their biosynthesis, transport, and regulation involve complex processes, specialized enzymatic machinery, and intricate regulatory networks. This review examines the multifaceted roles of metallophores in microbial ecology and their potential applications in sustainable agriculture, emphasizing their key role in trace metal acquisition, nutrient cycling, and plant–microbe interactions. Furthermore, it explores the potential applications of metallophores in agriculture, bioremediation, and biotechnology, connecting their potential to the development of novel strategies for sustainable agriculture.

Recurrent urinary tract infection (rUTI) is a significant public health problem in women. General measures to prevent recurrence include behavioral changes and increased fluid intake, cranberry ingest, use of methenamine hippurate, antibiotic prophylaxis, D-mannose, probiotics, or vaccines. We conducted a literature review of the latest updates on preventing rUTI in December 2024. The search concluded with 27 articles that fulfilled our inclusion criteria. Our review demonstrated that behavioral changes such as correct genital hygiene, avoiding postponing micturition or defecation, urinating after sexual intercourse, and ingesting 1.5–2 L of water could prevent rUTI. The ingestion of cranberries reduces the risk of symptomatic, culture-verified urinary tract infections in women with rUTIs. Methenamine hippurate is an alternative to antibiotics to avoid rUTI. Estrogen reduces rUTI in women with hypoestrogenism. Limited evidence supports using D-mannose, probiotics, and vaccines to prevent rUTI. In conclusion, after successful treatment of the acute episode, preventative measures are needed to reduce rUTI frequency and morbidity according to each patient’s characteristics and preferences.