Canadian journal of microbiology最新文献

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 micro-organisms that reside inside tissues and considered that 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 (ITS) 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), was 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.

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.

Whole-genome sequence-based surveillance of bacteria for determinants of antimicrobial resistance 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 antimicrobial resistance determinants in bacteria sampled from food products and animals related to food production.

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. S. scabies 87.22 showed higher growth in the suberin supplemented medium than the two other strains. When co-cultured on 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.

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.

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 bla_ctx-m, bla_tem, and bla_shv, identified in 14 coconut and 2 mango samples, were further characterized by whole-genome sequencing and antimicrobial susceptibility testing. The most common gene was bla_ctx-m-15, detected in 20 unique strains. Two carbapenemase-producing strains were isolated from coconut: Enterobacter roggenkampii encoding bla_ndm-1 and Escherichia coli encoding bla_ndm-5. Subsequent quantitative PCR (qPCR) analysis of enrichments for bla_ctx-m/bla_ndm 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.

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.

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.

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.