Canadian Journal of Microbiology

The Cryptococcus neoformans and Cryptococcus gattii species complexes are the etiological agents of cryptococcosis, a disease responsible for 181 000 deaths annually worldwide due to late diagnosis and limited treatment options. Studies focusing on the identification of new substances with antifungal activity, such as essential oils (EOs), are urgently needed. While the antifungal effects of EO have already been suggested, their mechanism of action at the molecular level still requires evaluation. In this work, we assessed the molecular changes induced by the exposure of Cryptococus neoformans (H99) and Cryptococcus deuterogatti (R265) to lavender essential oil (LEO) using a morphological and proteomics approach. The identified proteins were categorized by Gene Ontology according to biological processes and molecular functions, and Kyoto Encyclopedia of Genes and Genomes pathway analysis was also conducted. Our findings indicate that LEO creates a stressful environment in both strains; however, the response to this stimulus differs between the two species. In C. neoformans, changes were observed in energy metabolism and pathways related to alternative sources of energy and oxidative stress response. In C. deuterogatti, changes were identified in pathways related to cellular architecture, implying that the cell underwent morphological changes such as membrane and cell wall stiffening.

The pathogenic Pseudomonas aeruginosa utilizes a quorum-sensing pathway for biofilm formation. The quorum-sensing proteins LasI and LasR of the Las system, alongside the swarming motility protein BswR, play a crucial role in the biofilm-mediated antibiotic resistance phenomenon. In this in-silico study, LasI, LasR, and BswR were the prime targets for binding studies by promising drug candidates like linalool, ferutinin, citronellal, and carvacrol. These monoterpenoid compounds are carefully considered for this study due to their reported anti-microbial activity. Among all, carvacrol exhibited the highest binding energies with LasI (-5.932 kcal/mol), LasR (-7.469 kcal/mol), and BswR (-4.42 kcal/mol). Furthermore, the MMGBSA scores between carvacrol and LasI, LasR, and BswR individually are -33.14 kcal/mol, -54.22 kcal/mol, and -41.86 kcal/mol, which further corroborated the strong binding. During 100ns of simulation, the ligand binds to the active sites of these proteins through the H-bonds at Ile107 of LasI, Tyr47 of LasR, and Leu57 of BswR. In addition, the RMSD values of the ligand-protein complex are within the appropriate range of less than 5Å. ADME/T analysis confirmed that carvacrol has the most negligible toxicity to mammalian cells. Hence, this finding is the first report to show that carvacrol can inhibit the Pseudomonas aeruginosa biofilms

Many arthropods, including economically important pests of stored grains, host intracellular bacterial symbionts. These symbionts can have diverse impacts on host morphology, stress tolerance, and reproductive success. The ability to rapidly determine the infection status of host insects and the identity of intracellular symbionts, if present, is vital to understanding the biology and ecology of these organisms. We used a microbiome profiling method based on amplicon sequencing to rapidly screen 35 captive insect colonies. This method effectively revealed single and mixed infections by intracellular bacterial symbionts, as well as the presence or absence of a dominant symbiont, when that was the case. Because no a priori decisions are required about probable host-symbiont pairing, this method is able to quickly identify novel associations. This work highlights the frequency of endosymbionts, indicates some unexpected pairings that should be investigated further, such as dominant bacterial taxa that are not among the canonical genera of endosymbionts, and reveals different colonies of the same host insect species that differ in the presence and identity of endosymbiotic bacteria.

Vibrio parahaemolyticus produces a key virulent factor known as thermostable direct hemolysin (TDH). TDH exhibits diverse biological activities, including hemolytic activity. The β-type hemolysis observed on Wagatsuma agar due to TDH is recognized as the Kanagawa phenomenon (KP). The tdh2 gene is primarily responsible for TDH production and the associated KP. AcsS was originally identified as an activator of swimming and swarming motility in V. parahaemolyticus. However, the extent of its potential to regulate other cellular pathways remains unclear. In this study, we explored the regulatory effects of AcsS on the hemolytic activity and tdh2 expression in V. parahaemolyticus. The data showed that V. parahaemolyticus hemolytic activity and tdh2 transcription were under the negative control of AcsS. Additionally, in vitro binding assays revealed that His-AcsS could not bind to the regulatory DNA region of tdh2. However, overexpression of AcsS in an Escherichia coli strain suppressed the expression of tdh2. Collectively, these results suggested that AcsS suppresses the hemolytic activity of V. parahaemolyticus through the downregulation of tdh2 transcription. The data enhanced our understanding of the regulatory networks governing tdh2 expression and the roles of AcsS in this bacterium.

This study characterized the genome of three Salmonella enterica serovar Heidelberg (S. Heidelberg) strains with different antimicrobial resistance (AMR) profile that were inoculated as a cocktail into fresh pine wood shavings (PWS). The strains were isolated from feces (SH-AAFC), carcass (SH-ARS), and thigh (SH-FSIS) of broiler chicken. SH-AAFC harbored an antimicrobial resistant gene (ARG) blaCMY-2 on an IncI1 plasmid while SH-FSIS harbored multiple ARGs (floR, cmlA1, tet(A), blaTEM-1B, ant(2″)-Ia, aph(6)-Id, aph(3″)-Ib, and sul2) on an IncC plasmid. SH-ARS was pan-susceptible. The die-off of Salmonella was determined at days 0, 1, 7, 14, and 21. Antibiotic susceptibility tests and whole genome sequencing were performed on 77 isolates. At 21 days post-inoculation, Salmonella abundance decreased by 4.4 Log10 CFU/g with the water activity of PWS being correlated with Salmonella survival. SH-AAFC clonal populations survived longer in PWS than SH-FSIS and SH-ARS populations. SH-AAFC clones persisting in litter carried higher copy number of Col plasmids than their ancestors, while some SH-ARS clones acquired a lysogenic bacteriophage from SH-FSIS populations. These results suggest that mobile genetic determinants such as plasmids (which could carry ARGs) and bacteriophage plays roles in the persistence of S. Heidelberg in the PWS used as broiler litter. Highlights S. Heidelberg survived up to 21 days in PWS which is often used as broiler bedding. S. Heidelberg abundance and survival was correlated with the water activity of PWS. S. Heidelberg strains that carried higher copy numbers of small Col plasmids were the dominant strains isolated from PWS at later time points. S. Heidelberg strains harboring transmissible plasmid carrying AmpC-like beta-lactamase gene persisted longer in PWS without antibiotic pressures for AMR.

Lake Guamuez is the second largest lake in Colombia and economically supports hundreds of families in the area. The main activities carried out in the region have focused on tourism, agriculture, livestock, and rainbow trout production; however, these activities have been associated with contamination of the lake. This research aimed to evaluate the water quality of Lake Guamuez using somatic coliphages (SCs) as bioindicators. For this purpose, periodic sampling was carried out for 6 months at nine strategic points of the lake. For the detection of SCs, the method described in 9211 D of the Standard Methods for the Examination of Water and Wastewater was used. The genomic variability and presence of virulence genes in the isolated SCs were determined. Water contamination in the lake is evident, and the SCs titer is greater in areas with a high flow of anthropogenic activities. An important degree of genetic diversity and a high prevalence of virulence genes could be observed among the SCs analyzed. The results when compared with guidelines and water quality standards from various countries showed concentrations of SCs higher than those allowed. The high prevalence of gastrointestinal diseases in the region suggests a link to water contamination.

This study aimed to evaluate the antifungal activities of cannabidiol (CBD) against C. albicans. Yeast cells were treated once or twice with different concentrations (from 0 to 20 mg/ml) of CBD, showing a significant (p < 0.05) decreased the growth of C. albicans, with cell concentrations ranging from 5.1 × 106 cell/mL in the control to 1.8 × 106 cell/mL after one exposure to 20 µg/mL CBD. This growth reduction was greater after two exposures to CBD. After two exposures to 20 µg/mL CBD, the cell concentration was only 1.1 × 106 cell/mL. Such a growth decrease in C. albicans was confirmed by a reduced number of CFUs and a lower MTT value compared to the control. The growth inhibition was supported by a significant (p < 0.001) decrease in the yeast-to-hyphae transition, ranging from 20 ± 0.2% in the control to 2 ± 0.5% after exposure to 20 µg/mL CBD. Biofilm formation was also significantly reduced following CBD exposure. CBD at 10 and 20 µg/mL promoted the death of C. albicans through an apoptosis/necrotic pathway. Altogether, our results suggest the possible use of CBD, a cannabis derivative, to control C. albicans infection, including oral candidiasis.

Aspergillus fumigatus is a globally distributed mold and a major cause of opportunistic infections in humans. Because most infections are from environmental exposure, it is critical to understand environmental populations of A. fumigatus. Soil is a major ecological niche for A. fumigatus. Here, we analyzed 748 soil isolates from 21 locations in six provinces and one territory in Canada. All isolates were genotyped using nine microsatellite markers. Due to small sample size and/or close proximities for some local samples, these isolates were grouped into 16 local geographic and ecological populations. Our results indicated high allelic and genotypic diversities within most local and provincial populations. Interestingly, low but statistically significant genetic differentiations were found among geographic populations within Canada, with relatively similar proportions of strains and genotypes belonging to two large genetic clusters. In Hamilton, Ontario, and Vancouver, BC, where two and three ecological populations were analyzed, respectively, we found limited genetic difference among them. Most local and provincial populations showed evidence of both clonality and recombination, with no population showing random recombination. Of the 748 soil isolates analyzed here, two were resistant to triazole antifungals. We discuss the implications of our results to the evolution and epidemiology of A. fumigatus.

Bacteria encounter various stressful conditions within a variety of dynamic environments, which they must overcome for survival. One way they achieve this is by developing phenotypic heterogeneity to introduce diversity within their population. Such distinct subpopulations can arise through endogenous fluctuations in regulatory components, wherein bacteria can express diverse phenotypes and switch between them, sometimes in a heritable and reversible manner. This switching may also lead to antigenic variation, enabling pathogenic bacteria to evade the host immune response. Therefore, phenotypic heterogeneity plays a significant role in microbial pathogenesis, immune evasion, antibiotic resistance, host niche tissue establishment, and environmental persistence. This heterogeneity can result from stochastic and responsive switches, as well as various genetic and epigenetic mechanisms. The development of phenotypic heterogeneity may create clonal populations that differ in their level of virulence, contribute to the formation of biofilms, and allow for antibiotic persistence within select morphological variants. This review delves into the current understanding of the molecular switching mechanisms underlying phenotypic heterogeneity, highlighting their roles in establishing infections caused by select bacterial pathogens.

Highly pathogenic avian influenza (HPAI) H5N1 has caused the deaths of more than 100 million birds since 2021, and human cases since 1997 have been associated with significant morbidity and mortality. Given recent detections of HPAI H5N1 in dairy cattle and H5N1 RNA detections in pasteurized retail milk in the United States, we established the pan-Canadian Milk Network in April 2024. Through our network of collaborators from across Canada, retail milk was procured longitudinally, approximately every 2 weeks, and sent to a central laboratory to test for the presence of influenza A virus RNA. Between 29 April and 17 July 2024, we tested 109 retail milk samples from all 10 Canadian provinces (NL, NS, PEI, NB, QC, ON, MB, SK, AB, and BC). All samples tested negative for influenza A virus RNA. This nationwide initiative was established for rapid retail milk screening as per the earliest reports of similar undertakings in the United States. Our independent testing results have aligned with reporting from federal retail milk testing initiatives. Despite no known HPAI infections of dairy cattle in Canada to date, H5N1 poses a significant threat to the health of both humans and other animals. By performing routine surveillance of retail milk on a national scale, we have shown that academic networks and initiatives can rapidly establish nationwide emerging infectious disease surveillance that is cost-effective, standardized, scalable, and easily accessible. Our network can serve as an early detection system to help inform containment and mitigation activities if positive samples are identified and can be readily reactivated should HPAI H5N1 or other emerging zoonotic viruses be identified in agricultural or livestock settings, including Canadian dairy cattle.

Rabies is a zoonotic infectious disease that targets the nervous system of human and animals and has about 100% fatality rate without treatment. Rabies virus is a bullet-like viral particle composed of five structural proteins, including nucleoprotein (N), phosphorylated protein (P), matrix protein (M), glycoprotein (G), and large subunit (L) of RNA-dependent RNA polymerase. These multifunctional viral proteins also play critical roles in the immune escape by inhibiting specific immune responses in the host, resulting in massive replication of the virus in the nervous system and abnormal behaviors of patients such as brain dysfunction and hydrophobia, which ultimately lead to the death of patients. Herein, the role of five structural proteins of rabies virus in the viral replication and immune escape and its implication for the development of vaccines were systemically reviewed, so as to shed light on the understanding of pathogenic mechanism of rabies virus.

Frankia represent a unique group of filamentous, sporangia-forming bacteria, renowned for their exceptional capacity to establish symbiotic partnerships with actinorhizal plants. The objective of this paper is to offer quantitative insights into the current state of frankia research and its future potential. A comprehensive bibliometric analysis covering the years 2000–2022 was conducted using Scopus and SciVal. A steady increase in both annual publication and international collaboration has been observed, particularly since 2013. Research performance metrics for the last 5 years (2018–2022) indicate China and India as leaders with high Field-Weighted Citation Impact scores. This analysis highlighted prominent authors, research groups, and the evolving research landscape, suggesting an increasing focus on molecular and genomic aspects. The genomic era has transformed our understanding of frankia biology, highlighting their significance in diverse ecological and agricultural contexts. This study comprehensively maps the evolving landscape of frankia research, emphasizing key milestones that have catalysed international interest in frankia–actinorhizal research, expanding our perception of frankia’s capabilities beyond its traditional symbiotic role. As research in this field progresses, a deeper comprehension of frankia–plant interactions, symbiotic signalling, and the intricacies of metabolic pathways holds the promise of revealing innovative techniques for optimizing nitrogen fixation and broadening the spectrum of host plants.

The change and mechanism of soil and soil bacterial diversity during the change of herbaceous litter thickness in desert areas is crucial to understand. In the study, the dominant herbaceous litter mixture in Baijitan National Nature Reserve was selected as the research material, and an experiment was established by adjusting the litter depth. The results showed that the measured values of soil physicochemical factors (total nitrogen, total protein, total potassium, available phosphorus, available potassium, pH, and soil water content) increased with the increase of herbaceous litter mixture thickness in 0–5 cm soil layer. Actinobacteria, Proteobacteria, Acidobacteria, Chloroflexi, and Gemmatimonadetes were the dominant bacterial phyla under different thicknesses of herbaceous litter in 0–5 and 5–10 cm soil layers. Balneimonas, Rubrobacter, and Geodermatophilus were the dominant bacterial genera under different thicknesses of herbaceous litter in 0–5 and 5–10 cm soil layers. There was no obvious change in the α-diversity index of bacterial community the same soil layer, but the α-diversity index in the 0–5 cm soil layer was lower compared to the 5–10 cm soil layer. The results of this study revealed that the change of herbaceous litter thickness had no significant effect on soil bacterial community structure in desert areas.

Mine environments in boreal and sub-boreal zones are expected to experience extreme weather events, increases in temperature, and shifts in precipitation patterns. Climate change impacts on geochemical stability of tailings contaminants and reclamation structures have been identified as important climate-related challenges to Canadian mining sector. Adapting current reclamation strategies for climate change will improve long-term efficiency and viability of mine tailings remediation/restoration strategies under a changing climate. Accordingly, mesocosm experiments were conducted to investigate associations of climate-driven shifts in microbial communities and functions with changes in the geochemistry of organic covers and underlying tailings. Our results show that warming appears to significantly reduce C:N of organic cover and promote infiltration of nitrogen into deeper, unoxidized strata of underlying tailings. We also observed an increase in the abundance of some nitrate reducers and sulfide oxidizers in microbial communities in underlying tailings. These results raise the concern that warming might trigger oxidation of sulfide minerals (linked to nitrate reduction) in deeper unoxidized strata where the oxygen has been eliminated. Therefore, it would be necessary to have monitoring programs to track functionality of covers in response to climate change conditions. These findings have implications for development of climate resilient mine tailings remediation/restoration strategies.

Klebsiella pneumoniae is a ubiquitous opportunistic pathogen of the family Enterobacteriaceae. K. pneumoniae is a member of the ESKAPEE pathogens (Enterococcus faecium, Staphylococcus aureus, K. pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., and Escherichia coli), a group of bacteria that cause nosocomial infections and are able to resist killing by commonly relied upon antimicrobial agents. The acquisition of antimicrobial resistance (AMR) genes is increasing among community and clinical isolates of K. pneumoniae, making K. pneumoniae a rising threat to human health. In addition to the increase in AMR, K. pneumoniae is also thought to disseminate AMR genes to other bacterial species. In this review, the known mechanisms of K. pneumoniae AMR will be described and the current state of AMR K. pneumoniae within Canada will be discussed, including the impact of the coronavirus disease-2019 pandemic, current perspectives, and outlook for the future.

The principal methods to maintain soil fertility in Sahel soils are largely allowing fields to go fallow and manure addition. These methods are not currently sufficient to improve soil fertility. To promote biological amendments, we aimed to understand the plant-growth promoting traits of various soil microbial isolates. The soils collected in different areas in Senegal exhibited a similar eDNA profile of bacteria; the dominant microbes were Firmicutes, followed by Proteobacteria and Actinobacteria. Of 17 isolates identified and tested, the vast majority solubilized rock phosphate and a large number grew on culture medium containing 6% salt, but very few degraded starches or hydrolysed carboxymethyl cellulose or produced siderophores. Upon single inoculation, Peribacillus asahii RC16 and Dietzia cinnamea 55 significantly increased pearl millet growth and yield parameters. For cowpea, plant shoot length was significantly increased by Pseudarthrobacter phenanthrenivorans MKAG7 co-inoculated with Bradyrhizobium elkanii 20TpCR5, and nearly all rhizobacteria tested significantly improved cowpea dry weight and pod weight. Additionally, the double inoculation of Dietzia cinnamea 55 and MKAG7 significantly increased shoot length, dry weight, and seed head weight of pearl millet. These isolates are promising inoculants because they are ecologically-friendly, cost-effective, sustainable, and have fewer negative effects on the soil and its inhabitants.

Plant growth promotion by Pseudomonas sp. 31-12 incorporated into a lettuce seed pelleting matrix was studied. We examined (1) the effect of five rhizosphere derived bacterial strains on green oak lettuce (Lactuca sativa L.) seed germination, root and shoot growth, as a strain selection step for seed coating and seed pelletizing studies, (2) population stability of Pseudomonas sp. 31-12 incorporated into a pelleting matrix on lettuce seed stored three months at 4 °C, and (3) lettuce growth promotion in the laboratory and greenhouse by Pseudomonas sp. 31-12 coated and pelletized seed. A spontaneous streptomycin mutant of Pseudomonas sp. 31-12 (str) was used to determine population size on seed and roots of 15- and 30-day-old lettuce. The population of Pseudomonas sp. 31-12str on coated and pelleted seed decreased from 10⁴ cfu/seed to 10³ cfu/seed after 3 months storage at 4 °C. However, the population exceeded 10⁴ cfu/g root dry mass and 10⁵/g root dry mass after 15 days and 30 days in the greenhouse. Leaf fresh mass was significantly increased (P ≤ 0.05) with Pseudomonas sp. 31-12 seed treatment as compared to noninoculated seed. In conclusion, pelletized lettuce seed with Pseudomonas sp. 31-12 promoted growth and yield in the greenhouse.

Less than 1% of native prairie lands remain in the United States. Located in eastern Washington, the rare habitat called Palouse prairie was largely converted to wheat monocropping. With this conversion came numerous physical, chemical, and biological changes to the soil that may ultimately contribute to reduced wheat yields. Here, we explored how wheat (Tritcum aestivum L.) seedling establishment, plant size, and heading, signifying the developmental transition to flowering, were affected by being planted in prairie soil versus agricultural soils. We then sought to understand whether the observed effects were the result of changes to the soil microbiota due to agricultural intensification. We found that prairie soil enhanced both the probability of wheat seedling survival and heading compared to agricultural soil; however, wheat growth was largely unaffected by soil source. We did not detect effects on wheat developmental transitions or phenotype when inoculated with prairie microbes compared with agricultural microbes, but we did observe general antagonistic effects of microbes on plant size, regardless of soil source. This work indicates that agricultural intensification has affected soils in a way that changes early seedling establishment and the timing of heading for wheat, but these effects may not be caused by microbes, and instead may be caused by soil nutrient conditions.

Antimicrobial resistance is an ever-increasing threat. The widespread usage of ciprofloxacin has led to the manifestation of resistance due to chromosomal mutations or the acquisition of plasmid-mediated quinolone resistance (PMQR) traits. Some particular PMQR traits, qnr genes, have been identified globally in clinical and environmental isolates. This study aimed to determine the prevalence of ciprofloxacin-resistant bacteria in aquatic environments in southern Ontario and investigate the extent of dissemination of ciprofloxacin resistance traits among the bacterial communities. We surveyed the prevalence of plasmid encoding qnr genes using a multiplex PCR assay of associated PMQR genes, qnrA, qnrB, and qnrS, on 202 isolates. Despite the absence of significant impacts on minimum inhibitory concentration levels, the presence of qnr genes correlates with heightened resistance to quinolones and nalidixic acid in some isolates. Taxonomic analysis highlights distinct differences in the composition and diversity of ciprofloxacin-sensitive (CipS) and ciprofloxacin-resistant (CipR) populations, with Proteobacteria dominating both groups. Importantly, CipR populations exhibit lower genetic diversity but higher prevalence of multiple antibiotic resistances, suggesting co-selection mechanisms. Co-occurrence analysis highlights significant associations between ciprofloxacin resistance and other antibiotic resistances, implicating complex genetic linkages. The results of our study signified the critical role of environmental monitoring in public health.

Boric acid is a broad-spectrum antimicrobial used to treat vulvovaginal candidiasis when patients relapse on the primary azole drug fluconazole. Candida albicans is the most common cause of vulvovaginal candidiasis, colloquially referred to as a “vaginal yeast infection”. Little is known about the propensity of C. albicans to develop BA resistance or tolerance (the ability of a subpopulation to grow slowly in high levels of drug). We evolved 96 replicates from eight diverse C. albicans strains to increasing BA concentrations to test the evolvability of BA resistance and tolerance. Replicate growth was individually assessed daily, with replicates passaged when they had reached an optical density consistent with exponential growth. Many replicates went extinct quickly. Although some replicates could grow in much higher levels of BA than the ancestral strains, evolved populations isolated from the highest terminal BA levels (after 11 weeks of passages) surprisingly showed only modest growth improvements and only at low levels of BA. No large increases in resistance or tolerance were observed in the evolved replicates. Overall, our findings illustrate that there may be evolutionary constraints limiting the emergence of BA resistance and tolerance, which could explain why it remains an effective treatment for recurrent yeast infections.

With antimicrobial resistance (AMR) rapidly evolving in pathogens, quick and accurate identification of genetic determinants of phenotypic resistance is essential for improving surveillance, stewardship, and clinical mitigation. Machine learning (ML) models show promise for AMR prediction in diagnostics but require a deep understanding of internal processes to use effectively. Our study utilised AMR gene, pangenomic, and predicted plasmid features from 647 Enterococcus faecium and Enterococcus faecalis genomes across the One Health continuum, along with corresponding resistance phenotypes, to develop interpretive ML classifiers. Vancomycin resistance could be predicted with 99% accuracy with AMR gene features, 98% with pangenome features, and 96% with plasmid clusters. Top pangenome features overlapped with the resistance genes of the vanA operon, which are often laterally transmitted via plasmids. Doxycycline resistance prediction achieved approximately 92% accuracy with pangenome features, with the top feature being elements of Tn916 conjugative transposon, a tet(M) carrier. Erythromycin resistance prediction models achieved about 90% accuracy, but top features were negatively correlated with resistance due to the confounding effect of population structure. This work demonstrates the importance of reviewing ML models’ features to discern biological relevance even when achieving high-performance metrics. Our workflow offers the potential to propose hypotheses for experimental testing, enhancing the understanding of AMR mechanisms, which are crucial for combating the AMR crisis.

The rise of vancomycin-resistant enterococci (VRE) due to antibiotic overuse poses a significant threat to long-term care patients and those with impaired immune systems. Therefore, it is imperative to seek alternatives to overcome multidrug resistance. This study aimed to evaluate totarol, a natural compound derived from Podocarpus totara, for its antibacterial activity against vancomycin-resistant Enterococcus faecalis (VREF). Totarol exhibited potent antibacterial activity at a very low concentration of 0.25 µg/mL and demonstrated antibiofilm effects through biofilm inhibitory concentration and biofilm eradication concentration assays. Confocal laser scanning microscopy confirmed that totarol inhibited not only biofilm mass but also bacterial cell viability. The combinatorial use of sublethal concentrations of totarol and vancomycin showed antibacterial activity, as observed in the time-kill assay. Quantitative polymerase chain reaction assays revealed a concentration-dependent downregulation of key virulence genes (vanA, ace, asa, efaA, and esp) in VREF when exposed to totarol. In summary, totarol emerges as a promising adjuvant with vancomycin for inhibiting VREF, addressing vancomycin resistance and biofilm formation—critical challenges associated with VRE infection. Since this was an in vitro study, the role of totarol in the clinical implications of VREF treatment remains to be demonstrated.

Salmonella enterica serovars are zoonotic bacterial that cause foodborne enteritis. Due to bacteria’s antibiotic resistance, using bacteriophages for biocontrol and treatment is a new therapeutic approach. In this study, we isolated, characterized, and analyzed the genome of vB_SenS_TUMS_E19 (E19), a broad host range Salmonella bacteriophage, and evaluated the influence of E19 on liquid eggs infected with Salmonella enterica serovar Enteritidis. Transmission electron microscopy showed that the isolated bacteriophage had a siphovirus morphotype. E19 showed rapid adsorption (92% in 5 min), a short latent period (18 min), a large burst size (156 PFU per cell), and a broad host range against different Salmonella enterica serovars. Whole-genome sequencing analysis indicated that the isolated phage had a 42 813 bp long genome with 49.8% G + C content. Neither tRNA genes nor those associated with antibiotic resistance, virulence factors, or lysogenic formation were detected in the genome. The efficacy of E19 was evaluated in liquid eggs inoculated with S. Enteritidis at 4 and 25 °C, and results showed that it could effectively eradicate S. Enteritidis in just 30 min and prevented its growth up to 72 h. Our findings indicate that E19 can be an alternative to a preservative to control Salmonella in food samples and help prevent and treat salmonellosis.

The rising antimicrobial resistance rates and declining antimicrobial discovery necessitate alternative strategies to combat multidrug-resistant pathogens. Targeting microbial virulence is an emerging area of interest. Traditionally, virulence factors were largely restricted to bacteria-derived toxins, adhesins, capsules, quorum sensing systems, secretion systems, factors required to sense, respond to, acquire, or synthesize, and utilize trace elements (such as iron and other metals) and micronutrients (such as vitamins), and other factors bacteria use to establish infection, form biofilms, or damage the host tissues and regulatory elements thereof. However, this traditional definition overlooks bacterial virulence that may be induced or influenced by host-produced metabolites or other chemicals that bacteria may encounter at the infection site. This review will discuss virulence from a non-traditional perspective, shedding light on chemical-mediated host–pathogen interactions and outlining currently available mechanistic insight into increased bacterial virulence in response to host factors. This review aims to define a possibly underestimated theme of chemically mediated host–pathogen interactions and encourage future validation and characterization of the contribution of host chemicals to microbial virulence in vivo. From this perspective, we discuss proposed antivirulence compounds and suggest new potential targets for antimicrobials that prevent chemical-mediated virulence. We also explore proposed host-targeting therapeutics reducing the level of host chemicals that induce microbial virulence, serving as virulence attenuators. Understanding the host chemical-mediated virulence may enable new antimicrobial solutions to fight multidrug-resistant pathogens.

The number of copies of each chromosome, or ploidy, of an organism is a major genomic factor affecting adaptation. We set out to determine how ploidy can impact the outcome of evolution, as well as the likelihood of evolutionary rescue, using short-term experiments with yeast (Saccharomyces cerevisiae) in a high concentration of the fungicide nystatin. In similar experiments using haploid yeast, the genetic changes underlying evolutionary rescue were highly repeatable, with all rescued lines containing a single mutation in the ergosterol biosynthetic pathway. All of these beneficial mutations were recessive, which led to the expectation that diploids would find alternative genetic routes to adaptation. To test this, we repeated the experiment using both haploid and diploid strains and found that diploid populations did not evolve resistance. Although diploids are able to adapt at the same rate as haploids to a lower, not fully inhibitory, concentration of nystatin, the present study suggests that diploids are limited in their ability to adapt to an inhibitory concentration of nystatin, while haploids may undergo evolutionary rescue. These results demonstrate that ploidy can tip the balance between adaptation and extinction when organisms face an extreme environmental change.