Development of droplet digital PCR assays to quantify genes involved in nitrification and denitrification, comparison with quantitative real-time PCR and validation of assays in vineyard soil
This study optimized eight ddPCR assays to quantify total bacteria and archaea as well as the nitrification (bacterial and archaeal amoA) and denitrification (nirS, nirK, nosZI, nosZII) genes involved in the generation or reduction of the greenhouse gas nitrous oxide. Detection and quantification thresholds were compared with those of quantitative real-time PCR and were equal to, or improved, in ddPCR. To validate the assays using environmental samples, soil DNA was isolated from two vineyards in the Okanagan valley in British Columbia, Canada, over the 2017 growing season. Soil properties related to the observed gene abun...
Source: Canadian Journal of Microbiology - September 10, 2020 Category: Microbiology Authors: Tanja M. Voegel Melissa M. Larrabee Louise M. Nelson Source Type: research
Phosphate efflux as a test of plasma membrane leakage in Saccharomyces cerevisiae cells
Canadian Journal of Microbiology, Ahead of Print. Plasma membrane integrity is a key to cell viability. Currently, the main approach to assessing plasma membrane integrity is the detection of penetration of special dyes, such as trypan blue and propidium iodide, into the cells. However, this method needs expensive equipment: a fluorescent microscope or a flow cytometer. Besides, staining with propidium iodide occasionally gives false-positive results. Here, we suggest the phosphate (Pi) leakage assay as an approach to assess the increase in permeability of the plasma membrane of yeast cells. We studied the dependence of ph...
Source: Canadian Journal of Microbiology - September 10, 2020 Category: Microbiology Authors: Ludmila Trilisenko Airat Valiakhmetov Tatiana Kulakovskaya Source Type: research
A bacteriophage infecting Mesorhizobium species has a prolate capsid and shows similarities to a family of Caulobacter crescentus phages
Canadian Journal of Microbiology, Ahead of Print. Mesorhizobium phage vB_MloS_Cp1R7A-A1 was isolated from soil planted with chickpea in Saskatchewan. It is dissimilar in sequence and morphology to previously described rhizobiophages. It is a B3 morphotype virus with a distinct prolate capsid and belongs to the tailed phage family Siphoviridae. Its genome has a GC content of 60.3% and 238 predicted genes. Putative functions were predicted for 57 genes, which include 27 tRNA genes with anticodons corresponding to 18 amino acids. This represents the highest number of tRNA genes reported yet in a rhizobiophage. The gene arrang...
Source: Canadian Journal of Microbiology - September 9, 2020 Category: Microbiology Authors: K.M. Damitha Gunathilake Anupama P. Halmillawewa Keith D. MacKenzie Benjamin J. Perry Christopher K. Yost Michael F. Hynes Source Type: research
Quantitative proteomic profiling of shake flask versus bioreactor growth reveals distinct responses of Agrobacterium tumefaciens for preparation in molecular pharming
Canadian Journal of Microbiology, Ahead of Print. The preparation of Agrobacterium tumefaciens cultures with strains encoding proteins intended for therapeutic or industrial purposes is an important activity prior to treatment of plants for transient expression of valuable protein products. The rising demand for biologic products such as these underscores the expansion of molecular pharming and warrants the need to produce transformed plants at an industrial scale. This requires large quantities of A. tumefaciens culture, which is challenging using traditional growth methods (e.g., shake flask). To overcome this limitation...
Source: Canadian Journal of Microbiology - August 26, 2020 Category: Microbiology Authors: N. Prudhomme C. Gianetto-Hill R. Pastora W.-F. Cheung E. Allen-Vercoe M.D. McLean D. Cossar J. Geddes-McAlister Source Type: research
Functional analysis of the ocnE gene involved in nicotine-degradation pathways in Ochrobactrum intermedium SCUEC4 and its enzymatic properties
In conclusion, the ocnE gene is responsible for the ability of 2,5-dihydroxypyridine dioxy genase. These findings will be beneficial in clarifying the mechanisms of nicotine degradation in O. intermedium SCUEC4. (Source: Canadian Journal of Microbiology)
Source: Canadian Journal of Microbiology - August 25, 2020 Category: Microbiology Authors: Meng-fei Yu Zhen-zhen Xia Jia-cheng Yao Zhe Feng Ding-hua Li Tao Liu Guo-jun Cheng Dong-lan He Xiao-hua Li Source Type: research
Evaluation of biochars as carriers for Rhizobium leguminosarum
Canadian Journal of Microbiology, Ahead of Print. Peat is the standard carrier material used for commercial microbial inoculants produced in Canada and the United States. Peat is a slowly renewable resource and its production is extremely vulnerable to variable weather conditions. Furthermore, it may not be widely available in all countries. We investigated the potential to develop biochar as a carrier material. Our goal was to evaluate if different biochars perform comparably in supporting rhizobial survival, and what characteristics contribute to their ability to support rhizobial survival. Evaluation included characteri...
Source: Canadian Journal of Microbiology - August 19, 2020 Category: Microbiology Authors: Kimberly Hardy J. Diane Knight Source Type: research
Rates of fluoroquinolone resistance in domestically acquired Campylobacter jejuni are increasing in people living within a model study location in Canada
Canadian Journal of Microbiology, Ahead of Print. Antimicrobial resistance was evaluated in Campylobacter jejuni isolated from 1291 diarrheic people over a 15-year period (2004 –2018) in southwestern Alberta, a model location in Canada with a high rate of campylobacteriosis. The prevalence of resistance to chloramphenicol, clindamycin, erythromycin, and gentamicin was low during the examination period (≤4.8%). Resistance to tetracycline remained consistently high (41.6 %–65.1%), and resistance was primarily conferred by plasmid-borne tetO (96.2%). Resistance rates to ciprofloxacin and nalidixic acid increased substan...
Source: Canadian Journal of Microbiology - August 17, 2020 Category: Microbiology Authors: G. Douglas Inglis Eduardo N. Taboada Valerie F. Boras Source Type: research
d-Amino acids in antimicrobial peptides: a potential approach to treat and combat antimicrobial resistance
Canadian Journal of Microbiology, Ahead of Print. Antimicrobial resistance is one of the leading challenges in the human healthcare segment. Advances in antimicrobial resistance have triggered exploration of natural alternatives to stabilize its seriousness. Antimicrobial peptides are small, positively charged oligopeptides that are as potent as commercially available antibiotics against a wide spectrum of organisms, such as Gram-positive bacteria, Gram-negative bacteria, viruses, and fungal strains. In addition to their antibiotic capabilities, these peptides possess anticancer activity, activate the immune response, and ...
Source: Canadian Journal of Microbiology - August 12, 2020 Category: Microbiology Authors: Shikha Kapil Vipasha Sharma Source Type: research
Fungal endophyte diversity in table grapes
In this study, we determined the fungal microbial endophyte diversity in North American table grapes found at a Winnipeg, Manitoba, market. The amplicon internal transcribed spacer (ITS) metagenomics approach was used to profile the fungal communities of the fruit endophyte microbiome of three table grape types. The data supported endophyte diversity in different table grapes, including possible bioactive, saprophytic, and pathogenic fungi. Culturable endophytes were isolated and identified by morphology and ITS amplicon sequencing. The majority of the isolated culturable strains included Alternaria spp. and Cladosporium s...
Source: Canadian Journal of Microbiology - August 10, 2020 Category: Microbiology Authors: Champa Wijekoon Zoe Quill Source Type: research
Fungal endophyte diversity in table grapes
In this study, we determined the fungal microbial endophyte diversity in North American table grapes found at a Winnipeg, Manitoba, market. The amplicon internal transcribed spacer (ITS) metagenomics approach was used to profile the fungal communities of the fruit endophyte microbiome of three table grape types. The data supported endophyte diversity in different table grapes, including possible bioactive, saprophytic, and pathogenic fungi. Culturable endophytes were isolated and identified by morphology and ITS amplicon sequencing. The majority of the isolated culturable strains included Alternaria spp. and Cladosporium s...
Source: Canadian Journal of Microbiology - August 10, 2020 Category: Microbiology Authors: Champa Wijekoon Zoe Quill Source Type: research
Lactobacillus rhamnosus cell-free extract targets virulence and antifungal drug resistance in Candida albicans
Canadian Journal of Microbiology, Ahead of Print. Candidiasis caused by multidrug-resistant Candida species continues to be difficult to eradicate. The use of live probiotic bacteria has gained a lot of interest in the treatment of candidiasis; however, whole-cell probiotic use can often be associated with a high risk of sepsis. Strategies manipulating cell-free methods using probiotic strains could lead to the development of novel antifungal solutions. Therefore, we evaluated the effect of three probiotic cell-free extracts (CFEs) on the growth, virulence traits, and drug efflux pumps in C. albicans. On the basis of its m...
Source: Canadian Journal of Microbiology - August 10, 2020 Category: Microbiology Authors: Yvonne Dube Amber Khan Musa Marimani Aijaz Ahmad Source Type: research
CcpN: a moonlighting protein regulating catabolite repression of gluconeogenic genes in Bacillus subtilis also affects cell length and interacts with DivIVA
We report the interaction of CcpN with DivIVA of B. subtilis as determined using bacterial two-hybrid and glutathione S-transferase pull-down assays. Insertional inactivation of CcpN leads to cell elongation and formation of straight chains of cells. These findings suggest that CcpN is a moonlighting protein involved in both gluconeogenesis and cell elongation. (Source: Canadian Journal of Microbiology)
Source: Canadian Journal of Microbiology - August 6, 2020 Category: Microbiology Authors: Kusum Sharma Taranum Sultana Tanya E.S. Dahms Jo-Anne R. Dillon Source Type: research
CcpN: a moonlighting protein regulating catabolite repression of gluconeogenic genes in Bacillus subtilis also affects cell length and interacts with DivIVA
We report the interaction of CcpN with DivIVA of B. subtilis as determined using bacterial two-hybrid and glutathione S-transferase pull-down assays. Insertional inactivation of CcpN leads to cell elongation and formation of straight chains of cells. These findings suggest that CcpN is a moonlighting protein involved in both gluconeogenesis and cell elongation. (Source: Canadian Journal of Microbiology)
Source: Canadian Journal of Microbiology - August 6, 2020 Category: Microbiology Authors: Kusum Sharma Taranum Sultana Tanya E.S. Dahms Jo-Anne R. Dillon Source Type: research
