Canadian Journal of Microbiology

Canadian Journal of Microbiology

Published by Canadian Science Publishing

Online ISSN: 0008-4166

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Print ISSN: 1480-3275

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101 reads in the past 30 days

Phylogenetic tree showing members of the genus Morchella, inferred from Bayesian analysis of the combined ITS, RPB1, RPB2, and TEF1 dataset. Morchella sp. Mel-25 was used to root the tree. The Mel-8 collections in this dataset have possibly been misidentified due to their phylogenetic positioning. Sequences generated in this paper appear in bold typeface.
Tryphan blue stained Leucadendron coniferum roots collected from underneath Morchella capensis sp. nov. fruiting bodies showing mycorrhizal activity. Scale bars at 100 µm (A and C) and 50 µm (B and D). Photos: B. Van der Merwe
Morchella capensis sp. nov. ascomata at varying stages of maturity, showing a range of phenotypic expressions. Photos: J. Williams (A and D) and B. Foster (B and C).
Scanning electron microscopy imaging of the ascospores and asci of Morchella capensis sp. nov. Photos: J. Theron.
Light microscopy images of the stipe hairs (A), asci (B), and ascospores (C) of Morchella capensis. (D) showing the female cone-bearing Leucadendron coniferum, and the typical Hangklip sand Fynbos environment where the Leucadendron and M. capensis are found (E). Scale bar at 50 µm (A) and 10 µm (C). Photos: B. Van der Merwe (A–C) and B. Foster (D–E).

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Morchella capensis sp. nov., the Fynbos morel—the first description of an endemic Morchella species from Africa

February 2025

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201 Reads

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J. Williams

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E. Le Roux

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Aims and scope


The Canadian Journal of Microbiology contains new research in the field of microbiology, including:

Applied microbiology and biotechnology Microbial ecology, microbial structure, physiology and metabolism Virology, genetics and molecular biology Infection and immunity Fungi and other eucaryotic protists Microbiological methods

Recent articles


Biocontrol agent characteristics and its use in Brazil to control Witches’ Broom disease in cocoa plantation caused by Moniliophthora perniciosa. (A) Microorganisms used as biocontrol agents (BCAs) and their mechanisms of action. (B) Trichoderma stromaticum conidia in rice compose the biofungicide Tricovab used in Brazil to control Witches’ Broom disease in cocoa plantation caused by Moniliophthora perniciosa. Illustration created using Microsoft PowerPoint Version2311 with free images from Flaticon.com.
Trichoderma mechanism of action. (A) Mycoparasitism: (1) peptides and other small molecules produced by the host fungi are (2) recognized by Grp1 receptors and nitrogen sensors, (3) they activate the MAPK pathway to (4) induce transcription factors (TF) in the nucleus and (5) increase the expression of membrane and cell-wall degrading enzymes and secondary metabolites responsible for (6) degrading the cell wall of the host fungus, inhibiting their mycelial growth, promoting nutrient uptake and Trichoderma survives. These enzymes are also used to (7) remodel the Trichoderma cell wall. (B) Effects of Trichoderma on plant immune system: molecules produced by Trichoderma, including microbe-associated molecular patterns (MAMPS) and effector molecules and effector molecules, are recognized by pattern recognition receptors (PRR) and intracellular receptors (IR). This recognition leads to the production of nitrogen and oxygen reactive species, which activate mitogen-activated protein kinases (MAPKs). MAPKs, in turn, induce various plant defense mechanisms. Figure created using Microsoft PowerPoint Version2311.
Trichodermosis cases and Trichoderma virulence factors. (A) Common sites of Trichoderma infections (dos Santos and dos Santos 2023). (B) The Trichoderma species used in agriculture and reported to cause Trichodermosis, and their respective mechanisms of virulence implicated in human infection and modulation of mammalian immune system. Illustration created using Microsoft PowerPoint Version2311 with free images from Flaticon.com. TLR, Toll-like receptors; CLR, C-type lectin receptors.
Trichoderma infection models. (A) Immunosuppressed OF-1 mice exhibited elevated mortality rates corresponding to higher doses of Trichoderma longibrachiatum inoculation (Paredes et al. 2016). (B) The model of skin infection in immunosuppressed ICR mice indicates the presence of lesions and inflammation (Zhang and Li 2022). (C) A hypothesized mechanism for Trichoderma infection: (1) the infection initiates upon skin damage; (2) conidia germination and hyphal growth ensue, leading to (3) tissue invasion; (4) proteases, mycotoxins, and secondary metabolites secreted by Trichoderma cause; (5) damage to the host cell membrane and tissue; fungal antigens and damage-associated molecular patterns (DAMPs) released by dead cells activate; (6) macrophages to phagocytose the fungus and recruit; (7) neutrophils to induce neutrophil extracellular trap (NET) formation for Trichoderma elimination and preventing of fungal dissemination; (8) the immune response and Trichoderma-induced damage contribute to necrotic regions in the tissue; (9) Trichoderma conidia present in the tissue inhibit leukocyte recruitment and may serve as a pathway to fungal dissemination through adjacent blood vessels in infected tissue. Illustration created using Microsoft PowerPoint Version2311 with free images from Flaticon.com.
Common virulence factors from Trichoderma species involved in biocontrol agent (BCA), mycoparasitism, and relationship to human infectionsa.
Lessons from the field: Trichoderma in agriculture and human health
  • Literature Review
  • Publisher preview available

April 2025

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14 Reads

The use of Trichoderma in agriculture as both a biocontrol agent and biofertilizer hinges on its ability to colonize the rhizosphere, promote plant growth, endure adverse environments, compete for space and nutrients, and produce enzymes and secondary metabolites to mycoparasitize and infect other fungus. In humans, Trichoderma exhibits the capacity to infect various bodily tissues, leading to Trichodermosis. There has been a notable increase in cases ranging from superficial to fatal, invasive, and disseminated infections, particularly among immunocompromised individuals. Trichoderma species employ diverse strategies to colonize and survive in various environments, infecting phytopathogens; however, the mechanisms and virulence factors contributing to human infections remain poorly understood. In this mini review, we provide a brief overview and contextualization of the virulence mechanisms employed by Trichoderma in parasitizing other fungi, as well as those implicated in modulating plant immunity and inducing human infections. Furthermore, we discuss the similarity of these virulence factors capable of modulating the mammalian immune system and their potential implications for human infection.


From Patterns to Prediction: Machine Learning and Antifungal Resistance Biomarker Discovery

April 2025

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3 Reads

Fungal pathogens significantly impact human health, agriculture, and ecosystems, with infections leading to high morbidity and mortality, especially among immunocompromised individuals. The increasing prevalence of antifungal resistance (AFR) exacerbates these challenges, limiting the effectiveness of current treatments. Identifying robust biomarkers associated AFR could accelerate targeted diagnosis, shorten decision time for treatment strategies, and improve patient health. This paper examines traditional avenues of AFR biomarker detection, contrasting them with the increasingly effective role of machine learning (ML) in advancing diagnostic and therapeutic strategies. The integration of ML with technologies such as mass spectrometry, molecular dynamics, and various omics-based approaches, often result in the discovery of diverse and novel resistance biomarkers. ML's capability to analyse complex data patterns enhances the identification of resistance biomarkers and potential drug targets, offering innovative solutions to AFR management. This paper highlights the importance of interdisciplinary approaches and continued innovation in leveraging ML to combat AFR, aiming for more effective and targeted treatments for fungal infections.


Survey of fungal endophytes in barley under Fusarium head blight infection

Fusarium head blight (FHB) is a devastating fungal disease caused by Fusarium graminearum, which affects barley (Hordeum vulgare L.) and other small cereal grains. Fungal endophytes are microorganisms that reside inside tissues and considered that they may have been involved in various roles of the plants. This study involved the comparison of fungal endophytes between “non-infected/clean” and “FHB-infected” barley genotypes in various tissues collected at different plant developmental stages and were grown under different conditions (i.e., greenhouse, research field, and FHB-field nursery). We hypothesized that fungal endophytes diversity and abundance may differ between plant tissues in various barley genotypes that were non-infected and FHB-infected. The 18S-internal transcribed spacer sequencing analysis revealed a greater number of fungal operational taxonomic units (OTUs) and endophyte species in FHB-infected barley compared to clean barley. A one-way ANOVA and Tukey's pairwise comparison test (p ≤ 0.05) were performed to test significant differences. Higher seed endophyte diversity was found in FHB-infected (120 OTUs) compared to non-infected (113 OTUs) harvested in 2021. The increase in diversity of endophytes that contributes to different roles in plant protection and defense, such as biocontrol agents, may prevent the growth of Fusarium species and decrease FHB-infection.


Engineering Conjugative Plasmids for Inducible Horizontal DNA Transfer

April 2025

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9 Reads

Rapidly developing microbial resistance to existing antimicrobials poses a growing threat to public health and global food security. Current chemical-based treatments target cells by inhibiting growth or metabolic function, but their effectiveness is diminishing. To address the growing antimicrobial resistance crisis, there is an urgent need for innovative therapies. Conjugative plasmids, a natural mechanism of horizontal gene transfer in bacteria, have been repurposed to deliver toxic genetic cargo to recipient cells, showing promise as next-generation antimicrobial agents. However, the ecological risks posed by unintended gene transfer require robust biocontainment strategies. In this study, we developed inducible conjugative plasmids to solve these challenges. Utilizing an arabinose-inducible promoter, we evaluated 13 plasmids with single essential gene deletions, identifying trbC and trbF as strong candidates for stringent regulation. These plasmids demonstrated inducibility in both cis and trans configurations, with induction resulting in up to a 5-log increase in conjugation efficiency compared to uninduced conditions. Although challenges such as reduced conjugation efficiency and promoter leakiness persist, this work establishes a foundation for the controlled transfer of plasmids, paving the way for safer and more effective antimicrobial technologies.


Diversity and distribution characteristics of myxobacteria in the rhizosphere and nonrhizosphere soils of the halophyte Haloxylon ammodendron in the high saline-alkaline Ebinur Lake Wetland

This study employed Illumina HiSeq high-throughput sequencing technology to analyze the V4–V5 regions of myxobacterial 16S rRNA in rhizosphere and nonrhizosphere soils of Haloxylon ammodendron in the saline-alkaline wetland of Ebinur Lake, with the aim of investigating the community structure and diversity of myxobacteria. Results indicated that myxobacterial communities in rhizosphere soils exhibited greater diversity and richness compared to nonrhizosphere soils. Soil physicochemical properties, particularly moisture content, were identified as key environmental factors influencing myxobacterial diversity. The halotolerant genus Haliangium was found to be predominant under saline-alkaline conditions. Additionally, myxobacteria demonstrated distinct ecological specificity and environmental adaptability between rhizosphere and nonrhizosphere soils. For example, the genus Enhygromyxa exhibited a negative correlation with soil moisture content in rhizosphere soils but a positive correlation with soil electrical conductivity in nonrhizosphere soils. Co-occurrence network analysis revealed complex interaction patterns among myxobacterial genera and other bacterial genera, with closer interactions observed in rhizosphere soils. This study highlights the importance of environmental factors in regulating microbial community structure and function in saline-alkaline wetlands, providing new insights into the ecological roles and interaction mechanisms of myxobacteria within the ecosystem.


An epidemiological framework for improving the accuracy of whole-genome sequence-based antimicrobial resistance surveillance in Salmonella

March 2025

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12 Reads

Whole-genome sequence-based surveillance of bacteria for determinants of antimicrobial resistance (AMR) promises many advantages over traditional, wet-lab approaches. However, adjustments to parameters used to identify genetic determinants from sequencing data can affect results and interpretation of the important determinants in circulation. Using a dataset of whole-genome sequences from 1633 isolates of Salmonella Heidelberg and S. Kentucky collected from surveillance of Canadian poultry production, we queried the genomic data using an in silico AMR detection tool, StarAMR, applying a range of parameter values required for the detection pipeline to test for differences in detection accuracy. We compared the results from each iteration to phenotypic antimicrobial susceptibility results, and generated estimates of sensitivity and specificity using regression models that controlled for the effects of multiple sampling events and variables, and interactions between covariates. Results from our analyses revealed small, yet significant effects of the input parameters on the sensitivity and specificity of the AMR detection tool, and these effects differed based on the serovar and drug class in question. Findings from this study may have implications for the incorporation of whole-genome sequence-based approaches to the surveillance of AMR determinants in bacteria sampled from food products and animals related to food production.


Interactions between common scab-inducing strains on potato suberin

March 2025

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31 Reads

Potato tuber periderm is armored with suberin, that consists of two domains, an aliphatic domain composed of fatty acid polyesters and an aromatic domain composed of cinnamic acids. Streptomyces scabies 87.22, a predominant causal agent of potato common scab, was compared for adaptation to tuber suberin with Streptomyces acidiscabies ATCC 49003 and Streptomyces turgidiscabies Car8 belonging to emerging pathogenic species. Streptomyces scabies 87.22 showed higher growth in the suberin supplemented medium than the two other strains. When co-cultured in a rich nutrient medium, S. acidiscabies ATCC 49003 produced the antibiotic oxanthromicin, which inhibited growth and mycelium development of the other strains. Exposure of S. scabies 87.22 and S. acidiscabies ATCC 49003 to suberin was accompanied by the secretion of enzymes degrading cellulose, hemicellulose, fatty acids, and glycerol derivatives. Compared to the two other strains, S. scabies 87.22 showed higher esterase activity in suberin-supplemented medium and strong induction of cellulase gene expression. Both S. acidiscabies ATCC 49003 and S. turgidiscabies Car8 exhibited a poor utilization of trans-ferulic and p-coumaric acids, suggesting almost no ability to degrade the aromatic moiety of suberin. This work suggests that S. scabies 87.22 is better adapted to the potato periderm degradation than the emerging pathogens. The elucidation of pathogenic Streptomyces strains interaction may contribute to the improvement of ecologically oriented agronomic strategies for common scab management.


Workflow of phenotypic and genotyping testing of ciprofloxacin non-susceptibility in Salmonella enterica isolates. CIPARS, Canadian Integrated Program for Antimicrobial Resistance Surveillance; CLSI, Canadian Laboratory Standards Institute; WGS, whole-genome sequencing. Created in BioRender. Lerminiaux, N. (2024) https://BioRender.com/l91d653.
Heatmap of gene proportions conferring non-susceptibility to ciprofloxacin by year and isolate source. White spaces indicate that the gene was not present in that year and source.
Mechanisms of ciprofloxacin non-susceptibility in Salmonella enterica isolates from human and food/animal sources (n = 2325).
Molecular epidemiology and in silico prediction of ciprofloxacin resistance in Salmonella enterica in Canada, 2017–2022

March 2025

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16 Reads

Ciprofloxacin is important for treatment of severe or invasive Salmonella infections in humans. As laboratories transition from phenotypic to genomics-based methods for determining ciprofloxacin non-susceptibility, it is important to define the correlation between genetic determinants of resistance and phenotypic outcomes. Here, we examined ciprofloxacin resistance mechanisms in Salmonella and tested the hypothesis that isolates containing only one mechanism had intermediate resistance while isolates containing two or more mechanisms had full resistance according to breakpoints from the Clinical Laboratory Standards Institute. Among 13 750 human and food/animal Salmonella enterica isolates, 2325 were predicted to be non-susceptible to ciprofloxacin using whole genome sequencing and Staramr. The most common mechanisms of resistance were mutations in gyrA (especially S83F and D87N/D87Y) and the qnrB19 allele. Only 28% of ciprofloxacin resistant isolates had two or more resistance mechanisms; the remainder contained only one mechanism. Of isolates with two or more mechanisms, only 63% were resistant. Thus, the number of genetic determinants of ciprofloxacin resistance in an isolate could not reliably differentiate the ciprofloxacin intermediate or resistant categories when using North American breakpoints. Predicting ciprofloxacin intermediate/resistant as a single non-susceptible category would facilitate global standardization of data to inform public health surveillance, treatment guidelines, and stewardship.


Stacked bar plots of abundance of mean trimmed mean M-value (TMM) normalized phyla read counts present in beef cattle, swine feces, and broilers cecal contents.
Bar plot comparisons of fold changes of mean phylum abundance (TMM-normalized) from conventional (CONV) and natural (NAT) production systems for each livestock species. Negative values represent natural enrichment, and positive values represent conventional enrichment. (ANCOM-BC test with Benjamin–Hochberg p value adjustments; q > 0.05 = ns, q < 0.05 = *; q < 0.01 = **; q < 0.001 = ***; q < 0.0001 = ****)
Boxplot comparisons of prevalent (>1%) ARG class TMM-normalized total abundances between conventional versus natural AMU practices, separated by livestock species. (ANCOM-BC test with Benjamini–Hochberg p value adjustments; q > 0.05 = ns)
Boxplot comparisons of prevalent (>1%) ARG class TMM-normalized total abundances between livestock species, separated by conventional versus natural AMU practices. (ANCOM-BC test with Benjamini-Hochberg p value adjustments; q > 0.05 = ns; q < 0.05 = *; q < 0.001 = ***; q < 0.0001 = ****)
Boxplot comparisons of prevalent (>1%) ARG groups TMM-normalized total abundances between conventional versus natural AMU practices, separated by livestock species and ARG class. (ANCOM-BC test with Benjamini-Hochberg p value adjustments; q > 0.05 = ns; q < 0.05 = *; q < 0.01 = **, q < 0.001 = ***)
Comparative metagenomics reveals limited differences in antimicrobial resistance gene abundance across conventional and natural livestock production systems

February 2025

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24 Reads

The livestock industry has been a source of concern in terms of antimicrobial resistance (AMR) development and spread, especially from a One Health perspective. Raising livestock without antimicrobials, so called natural (NAT) production, is an increasingly popular practice. This study used metagenomics to compare this practice to conventional (CONV) antimicrobial use (AMU) on the microbiome and resistome in the feces of beef cattle and swine and the cecal contents of broiler chickens. In cattle, Bacteroidetes, Euryarchaeota, and Spirochaetes were more abundant (q < 0.01) in CONV than NAT systems, with no differences (q > 0.05) in bacterial profiles in either swine or chickens. Classes of antimicrobial resistant genes (ARG) were not impacted regardless of AMU in any of the livestock species. However, many tetracycline resistance genes were more abundant in CONV as compared to NAT swine (q < 0.05), but this difference was not observed in cattle or chickens. This study confirmed that elimination of AMU does not necessarily result in an immediate decline in the abundance or diversity of ARGs within a single livestock production cycle.


Morchella capensis sp. nov., the Fynbos morel—the first description of an endemic Morchella species from Africa

February 2025

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201 Reads

A species of Morchella was observed growing in spring, under a vulnerable member of the Proteaceae, in the Cape Floristic Region of South Africa. These fungi shared many of the cryptic characteristics common in the genus Morchella and displayed a wide range of phenotypic expression. The unique ecology of these fungi and the fact that no endemic Morchella species have been described from Africa lead to suspicions that this could be a novel species. Sequencing of key genetic regions, phylogenetics, and morphological studies confirmed that this was indeed a previously unknown species of Morchella. Roots collected underneath the fruiting bodies displayed a range of root-associated activities, alluding to a possible relationship. Further, this Morchella species has a history of traditional use on the Cape Peninsula of South Africa. The traditional use of fungi is rarely recorded in Africa. In this study, we introduce Morchella capensis sp. nov., the first endemic African morel.


Phylogenetic analyses comparing 063-A-02 to human clinical ST1/RT027 CA-CDI. (A) Minimum spanning tree using 2147 cgMLST alleles, calculated pairwise ignoring missing values, and presented on a logarithmic scale. Clostridioides difficile from retail meat is outlined in red. Isolates with a unique complex type are grouped as “Other”. (B) Unrooted maximum likelihood tree of SNV analysis of the cgMLST cluster containing complex type 5177. 94.33% of the core genome was used in the analysis and 98 variant sites were used to generate the phylogeny.
Phylogenetic analyses comparing 169-A-04 to human clinical ST10/RT015 CA-CDI. (A) Minimum spanning tree using 2147 cgMLST alleles, calculated pairwise ignoring missing values, and presented on a logarithmic scale. Grey highlight indicates clusters with ≤6 allele differences. Clostridioides difficile from retail meat is outlined in red. Isolates with a unique or unidentified complex type are grouped as “Other”. (B) Unrooted maximum likelihood tree of SNV analysis of the cgMLST cluster containing complex type 5179. 90.64% of the core genome was used in the analysis and 80 variant sites were used to generate the phylogeny.
Phylogenetic analyses comparing 180-A-01 to human clinical ST8/RT002 CA-CDI isolates. (A) Minimum spanning tree using 2147 cgMLST alleles, calculated pairwise ignoring missing values, and presented on a logarithmic scale. Grey highlight indicates a cluster with ≤6 allele differences. Clostridioides difficile from retail meat is outlined in red. Clinical isolates with a unique complex type are grouped as “Other”. (B) Unrooted maximum likelihood tree of SNV analysis including isolates with ≤6 cgMLST allele differences with the retail pork isolate. 90.00% of the core genome was used in the analysis and 173 variant sites were used to generate the phylogeny.
Antimicrobial susceptibilities (%) of Clostridioides difficile isolated from retail meat and human clinical infections.
Surveillance of Clostridioides difficile in Canadian retail meat and genomic linkages to community-associated human clinical infections in Canada

February 2025

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35 Reads

Community-associated Clostridioides difficile infections (CA-CDI) remain a concern in Canada, comprising a quarter of cases previously reported through the Canadian Nosocomial Infection Surveillance Program. Previous Canadian studies have reported toxigenic C. difficile isolated from Canadian retail meat, suggesting that it may be a source of exposure for CA-CDI in Canada. In this study, 3/219 (1.4%) of retail pork and 0/99 (0%) of retail beef samples tested positive for toxigenic C. difficile, which were molecularly characterized by PCR ribotyping and whole-genome sequencing. All three isolates were obtained from pork and belonged to sequence types (STs)/ribotypes (RTs) that have previously been isolated from human clinical CA-CDI cases in Canada: ST1/RT027, ST8/RT002, and ST10/RT015. Retail meat isolates were susceptible to the antimicrobials tested, save one isolate with intermediate resistance to clindamycin. Genomic comparison to Canadian human clinical CA-CDI isolates with the same corresponding ST/RT types showed two of the three pork isolates clustered with CA-CDI isolates via core-genome multilocus sequencing typing, with single nucleotide variant (SNV) analysis showing further genomic relatedness of 2-11 SNVs. Retail meat may therefore be a low source of CA-CDI exposure in Canada, with the potential for foodborne transmission of select clones.


Development and evaluation of a sensitive approach for detection and recovery of third-generation cephalosporin- and carbapenem-resistant Enterobacterales from ready-to-eat frozen stone fruit

February 2025

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33 Reads

Antimicrobial resistance (AMR) is a global public health threat, but the role of foods in its dissemination is poorly understood. We examined the incidence of foodborne bacteria carrying AMR genes considered high-priority research targets by the World Health Organization. Frozen, ready-to-eat, avocado, coconut, mango, and peach (n = 161) were tested for bacteria encoding extended-spectrum β-lactamases (ESBLs) and carbapenemases. Over 600 presumptive-positive isolates were recovered and analyzed with a pooled sequencing (Pool-seq) strategy. Coconut samples exhibited the highest bacterial loads and prevalence/diversity of AMR genes. Isolates harbouring the β-lactamase genes blactx-m, blatem, and blashv, identified in 14 coconut and 2 mango samples, were further characterized by whole-genome sequencing and antimicrobial susceptibility testing. The most common gene was blactx-m-15, detected in 20 unique strains. Two carbapenemase-producing strains were isolated from coconut: Enterobacter roggenkampii encoding blandm-1 and Escherichia coli encoding blandm-5. Subsequent quantitative PCR (qPCR) analysis of enrichments for blactx-m/blandm indicated a potentially higher prevalence of these genes than observed by colony screening. This study presents a practical method for recovering ESBL- and carbapenemase-producing bacteria from foods. Mapping their distribution in food products is crucial to assessing the role of foods in the global spread of AMR and developing effective public health interventions.


Cecal microbiome in broiler chicken related to antimicrobial feeding and bird’s sex

February 2025

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13 Reads

This study investigated the cecal microbiome of broilers raised under specific antimicrobial feeding programs (AFPs). A total of 2304 day-old Ross-708 male (M, n = 1152) and female (F, n = 1152) chicks were distributed into 48 floor pens which were allocated to one of three AFPs: Conventional, raised without medically important antibiotics (RWMIA), and raised without antibiotics (RWA). At 28 (D28) and 41 (D41) days of age, cecal contents were collected for culture dependent and independent analyses. At both 28 and 41 days, Enterococcus was more abundant in RWA-raised broilers than other groups with the most abundance of this bacterium being found in female birds (P < 0.05). At D41, the most abundant Eimeria tenella counts was observed in RWA-raised broiler ceca (P < 0.05). Sex effects were observed on the abundances of four of the 248 identified antimicrobial resistance genes while abundances of 10 were modulated by AFPs (P < 0.05). Ceca of females birds showed more tssB than males, and ceca of RWMIA-raised birds contained the highest abundance of chuY genes regardless of sex. This study showed that in a specific feeding program, cecal resistome can be affected by chicken’s sex contributing to understand the AMR related to the AMU.


Diagram of vector modules in the expanded BEVA2.0 vector archive. Dashed vector backbones reflect modules from BEVA1.0 (Geddes et al. 2019). Solid vector backbones reflect BEVA2.0 expansion modules presented in this work. Colored rounded rectangles reflect parts, colored by module type. Colored circles reflect sticky overhangs generated by BsmB1 and ligated by DNA ligase in Golden Gate cloning reactions. Plasmid names are indicated on the backbone with appropriate bacterial expression vector archive nomenclature below.
BEVA2.0 Broad Host Range Vectors. (A) Plasmid maps of broad host range vectors developed using BEVA2.0 parts. (B) Quantification of pBBR1 or RSF1010 plasmid maintenance with and with-out par loci. Data is expressed as the proportion of 100 colonies that grew on LB + Gm plates after patching following antibiotic-free culturing and spread-plating on LB. (C) Histograms of GFP signal from flow cytometry analysis of Escherichia coli ST18ALA bearing RSF1010, pBBR1, or RK2 plasmids expressing sfGFP. (D) Plasmid map of a curable sacB-containing pBBR1 plasmid pNDGG070, and the proportion of plasmid loss following growth on 10% sucrose based on sensitivity when 50 colonies were patched on LB + Sp media.
BEVA2.0 Genome Manipulation Plasmids. (A) Maps of plasmids developed for the double homologous recombination deletion method, and (B) schematic of the use of the plasmids to delete a target region. Steps involve: (i) cloning of homologous regions to the left and right of the target region into a BEVA2.0 vector using BsaI or BpiI Golden Gate cloning; (ii) conjugating the plasmid into the target organism and selecting for integration via single cross-over homologous recombination (using antibiotic resistance on the plasmid); (iii) introducing a replicating plasmid expressing the I-SceI endonuclease (e.g., via pDA1-ISceI-sacB) by conjugation and selecting for its maintenance to select for the second recombination event, in which the BEVA2.0 plasmid backbone is excised from the genome together with the target region; and (iv) screening resulting colonies for the presence of the deletion (based on PCR and loss of the resistance of the BEVA2.0 plasmid). (C) Maps of plasmids developed for Flp/FRT recombination methods, and (D) a schematic diagramming their use to excise and capture a target region. Steps involve: (i) cloning homologous regions to the left and right of the target adjacent to FRT site in heterologous BEVA2.0 plasmids via BsaI Golden Gate cloning; (ii) sequentially integrating the plasmids by conjugating them into the recipient and selecting for the appropriate antibiotic resistances; (iii) introducing a replicating plasmid containing flp recombinase gene (e.g., pTH2505) by conjugation and inducing Flp expression to catalyze excision of the target region via FRT recombination (optionally also introducing an E. coli recipient strain for region capture, and selecting with an alternate media that selects for the recipient with BEVA2.0 plasmids); and (iv) screening for the resulting deletion by PCR and loss of BEVA2.0 backbone resistance.
Parts to link bacterial expression vector archive (BEVA) and CIDAR. (A) Diagram of the architecture of BEVA Level 0 parts for open reading frame construction (Geddes et al. 2019). (B) Diagram of the architecture of CIDAR parts for open reading frame construction. (C) Architecture of a the BsaI multiple cloning site in Level 1 BEVA vectors. (D) Architecture of BEVA/standard European vector architecture “linker” DF terminators. (E) Fluorescence of BEVA vectors with CIDAR parts. The constitutive promoter J23106 together with the medium-strength RBS BCS12, are used to drive sfCFP (E), sfGFP (F), sfYFP (G) and mScarlet-I (H). The new linker terminator was used to assemble these marker genes in the RK2 BEVA2.0 plasmid pNDGG003. Fluorescence of the cloned plasmids in Escherichia coli (pNDMS054-57) is shown next to the empty vector control (pNDGG003) on the left side of the graphs. On the right side of the graphs, Sinorhizobium meliloti RmP110 containing pNDGG003 (RmND113) is shown compared to RmP110 containing pNDMS054-57 (RmND109-112).
Plasmids used in this work.
BEVA2.0: modular assembly of golden gate-compatible vectors with expanded utility for genetic engineering

February 2025

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31 Reads

This expansion for the modular vector assembly platform BEVA (Bacterial Expression Vector Archive) introduces 11 new BEVA parts including two new cloning site variants, two new antibiotic resistance modules, three new origins of replication, and four new accessary modules. As a result, the modular system is now doubled in size and expanded in its capacity to produce diverse replicating plasmids. Furthermore, it is now amenable to genetic engineering methods involving genome-manipulation of target strains through deletions or integrations. In addition to introducing the new modules, we provide several BEVA-derived Golden Gate cloning plasmids that are used to validate parts and that may be useful for genetic engineering of proteobacteria and other bacteria. We also introduce new parts to allow compatibility with the CIDAR MoClo parts libraries.


The application of shrub willow chip organic amendments impacts soil microbial community dynamics

February 2025

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35 Reads

Incorporating shrub willow chips into soil may improve the chemical, physical, and biological properties of soils with low organic matter but the impact on soil microbial communities and their dynamics is not known. We assessed changes in the soil microbial communities in response to willow chip applied at increasing rates (0, 20, 40, and 60 Mg ha⁻¹) in a potato-barley cropping system. Bacterial and fungal community diversity, relative abundance, and potential functions were assessed using amplicon sequencing of 16S and ITS rRNA genes at six time points. High rates (40 and 60 Mg ha⁻¹) of willow chips had no effect on bacterial alpha diversity but significantly decreased fungal alpha diversity (Shannon) while increasing fungal richness (Chao-1). At rates of 40 Mg ha⁻¹ and higher, the relative abundance of copiotrophic bacterial groups increased, while that of copiotrophic fungal groups decreased. The relative abundance of the most dominant microbial phyla and genera varied over time, with copiotrophic groups declining and oligotrophic groups increasing. High willow chip application rates increased bacterial molecular markers related to carbon fixation and degradation, nitrogen fixation, and phosphorus solubilization, while decreasing markers related to cellobiose transport and denitrification. This study demonstrates the ability of willow chips to influence the microbial community composition and potential function over time.


Assessment of viable Enterobacteriaceae and Enterococcus spp. across the agro-ecosystem of an antibiotic-free swine farm

January 2025

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23 Reads

Agricultural practices, specifically the use of antibiotics and other biocides, have repercussions on human, animal, and plant health. The aim of this study was to evaluate the levels of Enterobacteriaceae and Enterococcus, as antibiotic-resistant marker bacteria, in various matrices across the agro-ecosystem of an antibiotic-free swine farm in Quebec (Canada), namely pig feed, feces, manure, agricultural soil, water and sediment from a crossing stream, and soil from nearby forests. Samples were collected in fall 2022, spring and fall 2023, and spring 2024. All samples were subjected to counts of total, cefotaxime-, and ciprofloxacin-resistant Enterobacteriaceae as well as total and vancomycin-resistant Enterococcus spp. The frequency of total and cefotaxime-resistant Enterobacteriaceae along with the vancomycin-resistant Enterococcus decreased with age in pig feces, from weaning to the end of the fattening period. High proportions of the Enterobacteriaceae recovered from feces and environmental samples were resistant to cefotaxime. Application of manure on fields contributed a significant input of Enterococcus, but those resistant to vancomycin were under the detection limit. This study shows the prevalence of antibiotic-resistant bacteria in a farm agro-ecosystem even without the administration of antibiotics to the animals and highlights the complexity of components influencing antimicrobial resistance in the environment.


Map of the Hudson Bay Marine System, which shows the 18 sampling sites. Sampling was conducted from east to west, starting in Nain, Newfoundland and Labrador, and ending in Churchill, Manitoba. The sea ice concentration over the sampling period is shown, derived from records complied for 25 July 2022 (±3 days). Open water is any sea ice concentration <15%, and full ice is any concentration >90%. Ice concentration was plotted using ArcMap v10.8.2 (ESRI 2011) with data from Canadian Ice Service (Canadian Ice Service 2022).
Analysis of microbial communities using NMDS ordination of weighted UniFrac distance matrices. Environmental variables are shown fitted to the ordination with vectors (arrows) scaled by their significance (p value). Statistically significant (p < 0.05) fitted environmental variables (salinity, temperature) are further identified with red labels. Point colour scale represents the salinity (A) and temperature (B) of each sample. NMDS, nonmetric multidimensional scaling.
Heatmap of the 20 most abundant ASVs present in the samples. Colour coding represents the family to which each ASV belongs, and the size of each point corresponds to its relative abundance. Sites are sorted by their geographic cluster (as shown in Fig. 5), with the cluster label for each group on the top x-axis. ASV, amplicon sequence variant.
Number of ASVs versus salinity (A) and temperature (B); Faith’s PD versus salinity (C) and temperature (D); and Shannon Index versus salinity (E) and temperature (F). Spearman’s rho and p values are listed on each graph. Colours represent the respective environmental variable, with salinity and temperature in °C. ASV, amplicon sequence variant.
Environmental conditions measured by the flow-through thermosalinograph overlayed on sampling sites, as shown for (A) weighted-grid average salinity and (B) weighted-grid average temperature. Sites where DNA was collected are shown in large dots, coloured by hierarchical clusters as shown in heatmap of ASVs present in three or more samples (C). Distinct clusters represent communities in the Labrador Current (cluster 1, purple), Foxe Strait/Hudson Bay outflow (cluster 3, green), Hudson Bay offshore waters (cluster 4, orange), and the Churchill Estuary (cluster 5, peach). ASV, amplicon sequence variant.
Microbial communities in the Hudson Strait amidst rapid environmental change

January 2025

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12 Reads

Climate change is rapidly altering Arctic marine environments, leading to warmer waters, increased river discharge, and accelerated sea ice melt. The Hudson Bay Marine System experiences the fastest rate of sea ice loss in the Canadian North resulting in a prolonged open water season during the summer months. We examined microbial communities in the Hudson Strait using high throughput 16s rRNA gene sequencing during the peak of summer, in which the bay was almost completely ice-free, and air temperatures were high. We found that salinity and temperature significantly affected the taxonomic composition among microbial communities across sites. We observed a higher relative abundance of specific Polaribacter sp. Amplicon sequence variants (ASVs) at more saline sites. Shannon diversity was not significantly impacted by salinity or temperature. These results contribute to our understanding of surface water microbial community composition in the Hudson Strait and shed light on how future salinity and temperature conditions may favour certain microbial populations.


Crossing the streams: improving data quality and integration across the One Health genomics continuum with data standards and implementation strategies

January 2025

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34 Reads

The Canadian Genomics Research and Development Initiative for Antimicrobial Resistance (GRDI-AMR) uses a genomics-based approach to understand how health care, food production and the environment contribute to the development of antimicrobial resistance. Integrating genomics contextual data streams across the One Health continuum is challenging because of the diversity in data scope, content and structure. To better enable data harmonization for analyses, a contextual data standard was developed. However, development of standards does not guarantee their use. Implementation strategies are critical for putting standards into practice. This work focuses on the development of implementation strategies to better operationalize data standards across the Canadian federal genomics ecosystem. Results include improved understanding of complex data models that can create challenges for existing systems. Technical implementation strategies included spreadsheet-based solutions, new exchange formats, and direct standards integration into new databases. Data curation exercises highlighted common data collection and sharing issues, which informed improved practices and evaluation procedures. These new practices are contributing to improved data quality and sharing within the GRDI-AMR consortium as evidenced by publicly available datasets. The implementation strategies and lessons learned described in this work are generalizable for other standards and can be applied more broadly within other initiatives.


A pilot study on the effects of in-feed probiotic Lactobacillus rhamnosus ATCC 53103 (LGG) on vaccinated Atlantic salmon (Salmo salar): microbiomes and Aeromonas salmonicida challenge resilience

January 2025

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41 Reads

The use of probiotics is an alternative approach to mitigate the proliferation of antimicrobial resistance in aquaculture. In our study, we examined the effects of Lactobacillus rhamnosus GG (ATCC 53103, LGG) delivered in-feed on the weight, length, skin mucus, and faecal microbiomes of Atlantic salmon. We also challenged the salmon with Aeromonas salmonicida 2004-05MF26 (Asal2004) and assessed the mortality. Our results showed no significant change (P > 0.05) in weight or length of Atlantic salmon or their resilience to Asal2004 infection after LGG feeding. Infection changed significantly the skin mucus and faecal microbiomes: Clostridium sensu stricto increased from 3.14% to 9.20% in skin mucus and 1.39% to 3.74% in faeces (P < 0.05). Aeromonas increased from 0.02% to 0.60% in faeces (P < 0.05). Photobacterium increased from not detected (0%) to 52.16% (P < 0.01) and Aliivibrio decreased from 67.21% to 0.71% in faeces (P < 0.01). After infection, Lactococcus (9.93%) and Lactobacillus (2.11%) in skin mucus of the LGG group were significantly higher (P < 0.05) than in the skin mucus from the rest of the groups (4.14% and 1.08%, respectively). In conclusion, LGG feeding did not further increase the resilience of vaccinated Atlantic salmon. Asal2004 infection had much greater impact on skin mucus and faecal microbiomes than LGG feeding.


Acinetobacter baumannii ATCC17978-VU growth and virulence in rich versus minimal media. Difference in growth (A) and virulence (B) of A. baumannii ATCC17978-VU in rich LB media compared to M9 minimal media supplemented with 20 mmol/L sodium succinate (M9Succ).
Volcano plot of differentially expressed genes in rich versus minimal media. Genes that are differentially expressed in Acinetobacter baumannii ATCC17978-VU grown in rich LB media compared to M9 minimal media supplemented with 20 mmol/L sodium succinate (M9Succ). Significance cut offs set to a p value of 0.05 and a log2 fold change of ±1.2.
Clusters of orthologous genes (COG) categories of differentially expressed genes in rich versus minimal media. Number of genes differentially expressed sorted by COG category in Acinetobacter baumannii ATCC17978-VU grown in rich LB media compared to M9 minimal media supplemented with 20 mmol/L sodium succinate (M9Succ). Significance cut offs set to a p value of 0.05 and a log2 fold change of ±1.2. Category “None” refers to genes unable to be placed in established COG categories.
Minimum inhibitory concentrations (MIC) of Acinetobacter baumannii ATCC17978-VU grown in rich LB media compared to M9 minimal media supplemented with 20 mmol/L sodium succinate (M9Succ).
Phenotypic and transcriptomic changes in Acinetobacter baumannii in rich and minimal growth mediums

Acinetobacter baumannii is an opportunistic pathogen that is often studied in commonly used rich media in laboratories worldwide. Due to the metabolic versatility of A. baumannii, it can be cultured in different growth mediums; however, this can lead to genotypic and phenotypic variations. In this study, we compared phenotypic and transcriptomic changes in A. baumannii ATCC17978-VU cultured in M9 minimal media supplemented with 20 mmol/L sodium succinate and rich lysogeny broth media. Phenotypically, growth was significantly slowed, virulence in Galleria mellonella was attenuated, and susceptibility to a variety of antibiotic classes was reduced when A. baumannii ATCC17978-VU was grown in minimal media versus rich media. Transcriptomic analysis showed differential regulation of >700 genes—including those associated with energy production and ribosomal function—when the two growth conditions were compared, with the majority of the upregulated genes seen in minimal media of unknown function. This study showed that culture media has a profound effect on the phenotype and cellular workings of a bacteria, highlighting the need for more studies of pathogens like A. baumannii ATCC17978-VU in minimal media.


Genomic characterization of pathotype diversity and drug resistance among generic Escherichia coli isolated from broiler chickens in Canada

January 2025

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28 Reads

Escherichia coli is a Gram-negative bacterium that is ubiquitous in animals and humans, with some strains capable of causing disease. The aim of this study was to perform a comparative genomic analysis of 2732 generic E. coli isolates that were recovered from poultry samples collected from six regions in Canada as part of the National Microbiological Baseline study in Broiler Chicken. Isolates were subjected to whole genome sequencing and a subset (1122/2732) were tested for phenotypic resistance to 15 antimicrobials. These E. coli isolates were highly diverse, representing 376 serotypes, 236 sequence types and 21 pathotypes, of which 19 were hybrid pathotypes. A high concordance (>85%) between resistance phenotype and the presence of antimicrobial resistance genes and point mutations (resistance determinants) was observed for 13/15 antimicrobials. Over 95% of the β-lactam, fluoroquinolone, and phenicol resistance genes were predicted to be plasmid-borne. The number of resistance determinants per genome was highest in Quebec, while resistance genes associated with β-lactam resistance were more frequently detected in isolates from British Columbia. Generic E. coli in Canadian poultry are highly diverse, can carry pathotype-associated virulence factors and resistance determinants of clinical significance with a risk of emerging into pathogenic strains.



Tolerance mechanisms and molecular epidemiology of reduced susceptibility to chlorhexidine digluconate in different species of the Acinetobacter baumannii complex

December 2024

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11 Reads

The objective of this study was to compare chlorhexidine digluconate and other antibiotics susceptibility of four species of theAcinetobacter baumannii complex, and further investigate the chlorhexidine digluconate (CHG) tolerance mechanisms and molecular epidemic characteristics. Of 889 A. baumannii complex isolates, A. baumannii, A. nosocomialis, A. pittii, and A. seifertii accounted for 84.2%, 10.9%, 3.4%, and 1.5%. Acinetobacter baumannii was generally resistant to all tested antibiotics, while other three species were commonly more susceptible; 92.1% (313/340) CHG-tolerant A. baumannii, 19.6% (19/97) CHG-tolerant A. nosocomialis, 3.3% (1/30) CHG-tolerant A. pittii, and 15.4% (2/13) CHG-tolerant A. seifertii were identified. Furthermore, compared to A. baumannii ATCC 19606, upregulated expression was found in qacEΔ1, fabI, and efflux pump encoding genes in CHG-tolerant A. baumannii, but the expression level of oprD was reduced. Additionally, only the expression level of fabI was increased in the CHG-tolerant A. nosocomialis, and the expression level of adeG was increased in the CHG-tolerant A. pittii and A. seifertii. Furthermore, CHG-tolerant A. baumannii may have a relatively high clonal correlation, the predominant sequence type of which was ST208 (90%, 36/40). It is rather necessary to identify specific species members among the A. baumannii complex for clinical treatment options and antibiotics resistance monitoring.


Variation in plasmid conjugation among nontyphoidal Salmonella enterica serovars

December 2024

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12 Reads

Conjugation is a complex phenomenon involving multiple plasmid, bacterial, and environmental factors. Here we describe an IncI1 plasmid encoding multidrug antibiotic resistance to aminoglycosides, sulfonamides, and third-generation cephalosporins. This plasmid is widespread geographically among animal, human, and environmental sectors. We present data on the transmissibility of this plasmid from Salmonella enterica ser. Kentucky into 40 strains of S. enterica (10 strains each from serovars Enteritidis, Heidelberg, Infantis, and Typhimurium). Thirty seven out of 40 strains were able to take up the plasmid. Rates of conjugation were variable between strains ranging from 10⁻⁸ to 10⁻⁴. Overall, serovars Enteritidis and Typhimurium demonstrated the highest rates of conjugation, followed by Heidelberg, and then Infantis. No relationships were observed between the recipient cell surface and rate of conjugation. Recipient cell numbers correlated positively with conjugation rate and strains with high conjugation rates had marginally but significantly higher growth parameters compared to strains that took up the plasmid at lower frequencies. Environmental conditions known to impact cell growth, such as temperature, nutrient availability, and the presence of antibiotics, had a modulating effect on conjugation. Collectively, these results will further understanding of plasmid transmission dynamics in Salmonella, which is a critical first step towards the development of mitigation strategies.



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