Timothy F. Booth's research while affiliated with National Microbiology Laboratory, Canada and other places

Publications (68)

Article
Background: Serological assays designed to detect SARS-CoV-2 antibodies are being used in serological surveys and other specialized applications. As a result, and to ensure that the outcomes of serological testing meet high quality standards, evaluations are required to assess the performance of these assays and the proficiency of laboratories per...
Article
Full-text available
Bacterial resistance to the antiseptic chlorhexidine (CHX), is a growing problem, recently shown to be caused by deleterious mutations to the phospholipid transport system component (mlaA) as well as efflux pump overexpression. Comparisons of CHX resistance mechanisms, such as porin deletions (ompCF), and over-expressed efflux pumps (acrB, qacE, ac...
Article
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Chlorhexidine (CHX) is an essential medicine used as a topical antiseptic in skin and oral healthcare treatments. The widespread use of CHX has increased concerns regarding the development of antiseptic resistance in Enterobacteria and its potential impact on cross-resistance to other antimicrobials. Similar to other cationic antiseptics, resistanc...
Article
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Outer membrane vesicles (OMVs) produced by Gram-negative bacteria are mediators of cell survival and pathogenesis by facilitating virulence factor dissemination and resistance to antimicrobials. Studies of OMV properties often focus on hypervesiculating Escherichia coli mutants that have increased OMV production when compared to their corresponding...
Article
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Biocides such as quaternary ammonium compounds (QACs) are potentially important contributors towards bacterial antimicrobial resistance development, however, their contributions are unclear due to a lack of internationally recognized biocide testing standards. Methods to detect QAC tolerance are limited to laborious traditional antimicrobial suscep...
Preprint
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Quaternary ammonium compounds (QACs) benzalkonium (BZK) and cetrimide (CET) are common disinfectants used to inhibit or eradicate Gram-negative bacteria in clinical and agricultural products. QAC tolerance in Escherichia coli and other Enterobacterales species can confer cross-resistance to various clinically used antibiotics, making it important t...
Article
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Enteroviruses are single-stranded positive-sense RNA viruses that primarily cause self-limiting gastrointestinal or respiratory illness. In some cases, these viruses can invade the central nervous system, causing life-threatening neurological diseases including encephalitis, meningitis and acute flaccid paralysis (AFP). As we near the global eradic...
Article
Auritidibacter ignavus is a Gram-stain-positive bacillus derived from otorrhea. Four strains derived from ear discharges in Canada and Switzerland, with features consistent with but distinguishable from Auritidibacter ignavus IMMIB L-1656T (accession number FN554542) by 16S rRNA gene sequencing (97.5 % similarity), were thought to represent a novel...
Chapter
Filoviruses are highly filamentous enveloped animal viruses that can cause severe haemorrhagic fevers. The filovirus ribonucleoprotein forms a highly organized double-layered helical nucleocapsid (NC) containing five different virally encoded proteins. The inner layer consists of NP, the RNA binding protein, complexed with the monopartite linear ge...
Article
Background: In August 2014, a large North American outbreak of enterovirus D68 (EV-D68) was observed in association with severe respiratory illnesses in children as well as a possible association with acute flaccid paralysis. The full spectrum of clinical illness associated with EV-D68 is not well known. Objective: To compare children positive and...
Article
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The Zika virus (ZIKV) epidemic is an ongoing public health concern. ZIKV is a flavivirus reported to be associated with microcephaly, and recent work in animal models demonstrates the ability of the virus to cross the placenta and affect fetal brain development. Recent findings suggest that the virus preferentially infects neural stem cells and the...
Article
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We present the structure of the surface Ebola virus (EBOV) trimeric glycoprotein (GP) spike at 11 Å resolution, in situ within the viral plasma membrane of purified virus particles. GP functions in cellular attachment, endosomal entry, and membrane fusion to initiate infection, and is a key therapeutic target. Nevertheless, only about half of the G...
Article
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Important roles of microRNAs (miRNAs) in regulating the host response during viral infection have begun to be defined. However, little is known about the functional roles of miRNAs within an in vivo acute viral encephalitis model. We therefore identified global changes in miRNA expression during acute herpes simplex virus type 1 (HSV-1) encephaliti...
Article
Acute flaccid paralysis (AFP) is notifiable in Canada with a differential diagnosis that includes a number of conditions. This analysis describes the epidemiology of AFP in Canadian youth less than 15 years old. Methods: Monthly active surveillance for AFP was conducted as part of the Canadian AFP Surveillance System. Results: From 1996 to 2014, 85...
Article
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Despite being an excellent tool for investigating ultrastructure, scanning electron microscopy (SEM) is less frequently used than transmission electron microscopy for microbes such as viruses or bacteria. Here we describe rapid methods that allow SEM imaging of fully hydrated, unfixed microbes without using conventional sample preparation methods....
Article
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Discrepancy in synaptic structural plasticity is one of the earliest manifestations of the neurodegenerative state. In prion diseases, a reduction in synapses and dendritic spine densities is observed during preclinical disease in neurons of the cortex and hippocampus. The underlying molecular mechanisms of these alterations have not been identifie...
Article
The poliovirus is very close to being eradicated from the world. To this end, the four main objectives of the World Health Organization's Polio Eradication & Endgame Strategic Plan 2013-2018 are to: detect and interrupt all poliovirus transmission; strengthen immunization systems and withdraw oral polio vaccine; contain poliovirus and certify inter...
Article
A widespread outbreak of enterovirus D68 (EV-D68) was detected in association with respiratory illness in children across Canada and the United States during the autumn of 2014. The majority of cases were mild, but some were associated with more severe illness requiring hospitalization; some of the cases also had neurological symptoms including par...
Article
Full-text available
Abstract Background: Enterovirus D68 (EV-D68) has been detected infrequently and has not been associated with severe disease in Canada. In the early fall of 2014, following an unusual case increase in the United States, clusters of EV-D68 among children and some adults manifesting severe symptoms were reported in Canada. Objective: To provide an in...
Chapter
Filoviruses are unusual in being filamentous animal viruses. Like other mononegaviruses, filoviruses have helical nucleocapsids (NCs), which are the most complex amongst this virus order, containing 5 proteins: L, NP, VP35, VP30 and VP24 along with the 18.9 kb monopartite, negative-sense RNA genome. The filovirus NC is also unique in that it forms...
Article
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Clostridium difficile are Gram-positive, spore forming anaerobic bacteria that are the leading cause of healthcare-associated diarrhea, usually associated with antibiotic usage. Metronidazole is currently the first-line treatment for mild to moderate C. difficile diarrhea however recurrence occurs at rates of 15-35%. There are few reports of C. dif...
Article
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Background We report the first multi-site rotavirus genotype analysis in Canada. Prior to this study, there was a dearth of rotavirus G and P genotyping data in Canada. Publically funded universal rotavirus vaccination in Canada started in 2011 and has been introduced by four provinces to date. Uptake of rotavirus vaccines in Canada prior to 2012 h...
Article
We identified the interaction between HBV X (HBx) protein and the oncogene AIB1 (amplified in breast cancer 1). A serine/proline motif (SSPSPS) in HBx was found to be required for the interaction. Two LXD motifs [LLXX(X)L, X means any amino acids], LLRNSL and LLDQLHTLL in AIB1 were also found to be involved in the HBx-AIB1 interaction. The HBx-AIB1...
Article
Administrative databases are often used to determine burden of rotavirus disease in emergency departments (EDs). Our objective was to describe rotavirus ED visits to include healthcare utilization pre- and postvisit to estimate true societal costs. During rotavirus seasons in 2007, 2008, and 2009, a convenience sample of children <3 years of age wi...
Data
Surface spike distribution in the Ebola VLP. Longitudinal Z-slices through the top and middle of the particle are shown, as well as the end-on view (A). The tomogram is shown as a shaded surface at a density threshold that indicates the spikes (B). The volume from one side of the tomogram has been extracted, and a red-blue color scheme shows the de...
Data
Tomographic slices of Ebola virus. Slices in “Z” of Ebola virus tomogram reveal the components of the virus and nucleocapsid (A). The insets show 2D averages of the nucleocapsid. The slice through the top of the virus reveals the envelope and glycoprotein spikes. The next slice cuts through the top of the nucleocapsid (NC) revealing the banding pat...
Data
Tomogram of Ebola virus. This movie shows slices in “Z” through a cryo-electron tomogram of a linear region of Ebola virus. The slices which are in the Z-plane pass back and forth through the virus showing various components such as the surface glycoprotein spikes and the nucleocapsid in the interior of the virus. (MOV)
Data
Extraction of Ebola nucleocapsid structure for sub-tomographic analysis. The tomogram of a linear region of the Ebola virus was used as the first reference for sub-tomogram analysis (A). When viewed along the helical axis (Y) or from the end perspectives (X,Z) the basic components are visible. The tomographic volume was also cylindrically masked al...
Data
High magnification images showing linear regions of Ebola virus. The diameter of EBOV is constant in linear regions of the virus containing a nucleocapsid (A). The viral filaments are not perfectly straight, and are often curved, complicating helical image processing. Glycoprotein spikes (GP), envelope (E), and nucleocapsid (NC) are all clearly vis...
Article
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Filoviruses, including Ebola virus, are unusual in being filamentous animal viruses. Structural data on the arrangement, stoichiometry and organisation of the component molecules of filoviruses has until now been lacking, partially due to the need to work under level 4 biological containment. The present study provides unique insights into the stru...
Data
Ebola virus spike distribution. This movie shows one surface of a cryo-electron tomogram of an Ebola virus-like particle, generated by expressing the VP40 and GP proteins. The structure rotates showing the distribution of spikes. The locations of individual spikes are identified by white spheres, and the reconstruction is replaced by a cylinder to...
Data
Representative low-magnification images of Ebola virus. Frozen hydrated virus is clearly visible with sections of the filamentous virus over both the support film and across the holes in the quantifoil film. Individual G1 (single genome copy) virus is circled in red, several sections containing a nucleocapsid are indicated by a blue arrowhead, and...
Data
3-D reconstruction of Ebola virus nucleocapsid. This movie shows the three-dimensional structure of the of the Ebola virus nucleocapsid as shaded surface representation. The surface is set at a density threshold which would include one copy of NP, VP24,VP30, VP35, and the RNA. The nucleocapsid rotates, and then is sliced through the Z-axis to show...
Data
3-D reconstruction of Ebola virus spike. This movie shows the three-dimensional structure of the of the Ebola virus GP spike. The structure rotates showing views from different angles, and indicates the location of the docked 3CSY.pdb [42] structure with the GP1 and GP2 domains and KZ52 antibody (purple). (MOV)
Data
Length analysis of “continuous” Ebola virus particles. The length of 2090 EBOV particles were measured using ImageJ [52]. The values in the “model length” column are based on multiples of the G1 mean length. The values in the in the “mean length” column were calculated directly from the data. Only full particles containing a continuously packaged n...
Data
Modeling of RNA in the Ebola nucleocapsid. Two previously determined atomic resolution structures of negative stranded RNA viruses (VSV (2GIC.pdb) [58], and RSV (2WJ8.pdb) [32]) were used to estimate the EBOV nucleocapsid length and number of nucleotides per nucleoprotein. Images of VSV (A) and RSV (B) are shown as a molecular surface with the prot...
Article
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We have determined the structure of the core capsid of an unusual variant of hepatitis B virus, genotype G (HBV/G) at 14Å resolution, using cryo-electron microscopy. The structure reveals surface features not present in the prototype HBV/A genotype. HBV/G is novel in that it has a unique 36-bp insertion downstream of the core gene start codon. This...
Data
Microscopy results show that the SARS-9b protein localizes in both cytoplasm as well as nucleus when expressed in transfected mammalian cells. Vero cells were transfected with either pEYFPN1-9b or pEYFPN1 alone. After 36 hrs, cells were fixed and mounted. Arrow in panel (i) indicates that some protein also enters into the nucleus. Panel (iv) shows...
Article
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Background 9b is an accessory protein of the SARS-CoV. It is a small protein of 98 amino acids and its structure has been solved recently. 9b is known to localize in the extra-nuclear region and has been postulated to possess a nuclear export signal (NES), however the role of NES in 9b functioning is not well understood. Principal Findings/Methodo...
Data
SARS-CoV 9b pulls down some specific proteins from nuclear extract of Vero cells. A. The pCDNA3.1/V5-His TOPO-9b was used for in-vitro transcription and translation. When ran on a 15% SDS-acrylamide gel, dried and processed by autoradiography, a band of approx. 17 kDa was seen on the autoradiogram (lane 2). Lane 1 shows the mock lysate. B. The TNT...
Article
Biological Sciences SymposiaTransmission Electron Microscopy of the Ebola virusArticle author querybeniac d [PubMed] [Google Scholar]melito p [PubMed] [Google Scholar]jones s [PubMed] [Google Scholar]booth t [PubMed] [Google Scholar]DR Beniaca1, PL Melitoa1, SM Jonesa1 and TF Bootha1a1 National Microbiology Laboratory, Canada
Article
of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.
Chapter
Coronavirus spikes have the largest mass of any known viral spike molecule. The spike is a type 1 viral fusion protein, a class of trimeric surface glycoprotein proteins from diverse viral families that share many common structural and functional characteristics. Fusion proteins are mainly responsible for host cell receptor recognition and subseque...
Article
of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008
Article
Ebola virus is a filovirus that causes hemorrhagic fever in humans and is associated with case fatality rates of up to 90%. The lack of therapeutic interventions in combination with the threat of weaponizing this organism has enhanced research investigations. The expression of key viral proteins and the production of virus-like particles in mammali...
Article
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Of 4,268 wild ducks sampled in Canada in 2005, real-time reverse transcriptase-PCR detected influenza A matrix protein (M1) gene sequence in 37% and H5 gene sequence in 5%. Mallards accounted for 61% of samples, 73% of M1-positive ducks, and 90% of H5-positive ducks. Ducks hatched in 2005 accounted for 80% of the sample.
Article
In June 2006, 150 wild common carp were sampled from Hamilton Harbour, Lake Ontario, Canada. Tissue pools consisting of kidney, spleen and encephalon were screened for viruses as a condition facilitating the export of live carp to France. Cytopathic effect (CPE), indicative of a viral infection, became evident after 8 days of incubation at 15 degre...
Article
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In summer 2003, a respiratory outbreak was investigated in British Columbia, during which nucleic acid tests and serology unexpectedly indicated reactivity for severe acute respiratory syndrome coronavirus (SARS-CoV). Cases at a care facility were epidemiologically characterized and sequentially investigated for conventional agents of respiratory i...
Article
of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005
Article
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Cellular prion protein (PrP(C)) is a glycophosphatidylinositol (GPI)-anchored protein, of unknown function, found in a number of tissues throughout the body, including several blood components of which platelets constitute the largest reservoir in humans. It is widely believed that a misfolded, protease-resistant form of PrP(C), PrP(Sc), is respons...
Article
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Severe acute respiratory syndrome (SARS) is characterized by a risk of nosocomial transmission; however, the risk of airborne transmission of SARS is unknown. During the Toronto outbreaks of SARS, we investigated environmental contamination in SARS units, by employing novel air sampling and conventional surface swabbing. Two polymerase chain reacti...
Article
The human coronavirus, associated with severe acute respiratory syndrome (SARS-CoV), was identified and molecularly characterized in 2003. Sequence analysis of the virus indicates that there is only 20% amino acid (aa) identity with known coronaviruses. Previous studies indicate that protein-protein interactions amongst various coronavirus proteins...
Article
Severe Acute Respiratory Syndrome (SARS), an emerging disease characterized by atypical pneumonia, has recently been attributed to a novel coronavirus. The genome of SARS Coronavirus (SARS-CoV) has recently been sequenced, and a number of genes identified, including that of the nucleocapsid protein (N). It is noted, however, that the N protein of S...
Article
Full-text available
An outbreak of severe acute respiratory syndrome (SARS) began in Canada in February 2003. The initial diagnosis of SARS was based on clinical and epidemiological criteria. During the outbreak, molecular and serologic tests for the SARS-associated coronavirus (SARS-CoV) became available. However, without a "gold standard," it was impossible to deter...
Article
Full-text available
We sequenced the 29,751-base genome of the severe acute respiratory syndrome (SARS)-associated coronavirus known as the Tor2 isolate. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses, including two human coronaviruses, HCoV-OC43 and HCoV-229E. Phylogenetic analysis of the predicted viral prot...
Article
Tang, P Louie, M Richardson, SE Smieja, M Simor, AE Jamieson, F Fearon, M Poutanen, SM Mazzulli, T Tellier, R Mahony, J Loeb, M Petrich, A Chernesky, M McGeer, A Low, DE Phillips, E Jones, S Bastien, N Li, Y Dick, D Grolla, A Fernando, L Booth, TF Henry, B Rachlis, AR Matukas, LM Rose, DB Lovinsky, R Walmsley, S Gold, WL Krajden, S

Citations

... Seropositivity was defined as an OD 490 ratio greater than one with the experimentally determined cut-off, which corresponds to the mean of 10 negative controls plus three times the standard deviation of the negative controls for each experiment. This anti-RBD IgG ELISA was validated on the Canadian National Microbiology Laboratory SARS-CoV-2 National Serology Panel (23,26). Absence of previous infection to SARS-CoV-2 was confirmed in the naïve cohort by performing an ELISA detecting specific IgG antibodies for the Nucleoprotein (anti-N) of SARS-CoV-2 and the anti-RBD ELISA before vaccination. ...
... CzcRS has been shown to not only effect expression of czcCBA but also the expression of the outer membrane porin gene oprD in P. aeruginosa [120] leading to increased resistance to a number of antibiotics. Indeed, porin loss is an important component in increased tolerance to several biocides such as chlorhexidine [127] or benzalkonium chloride [67] as well as for several antibiotics [128]. The transcriptional activators SoxS, RobA and RamA are triggered under general stress conditions, which in turn leads to increased acrAB-TolC expression [129]. ...
... as compared to CHXR1, is likely due to the presence of other gene mutations in the adapted CHXR1 isolate as noted in its recent multi-omic analysis study 22 . In CHXR1, fimE, gadE, and cdaR all demonstrated minor (twofold) CHX MIC changes when they were either individually complemented in CHXR1 or they were examined as single E. coli gene deletion mutants 22 . ...
... The difficult separation and purification processes necessary to get significant amounts of these microscopic vesicular structures are one of the primary challenges of investigating OMVs. The majority of investigations identify ultracentrifugation and ultrafiltration as techniques [192]. Notably, the isolation process can impact the shape and yield of OMVs, increase OMV aggregation, and/ or collect lipoproteins and other undesirable cell debris. ...
... All previous RFDMIA analyses used stationary phase cells (Fig. 1). Stationary phase cells were selected for their speed and accuracy in past RFDMIA after a comparison of stationary and mid-log phase cells 28 . AcrAB and AceI are both secondary active proton motive force driven pumps that are most active when cells are growing with a respirable carbon source 19,40 . ...
... Once these viruses enter the interstitial matrix and drain into the lymphatic system, they become circulating viruses, and can then access the regional lymph nodes and disseminate to organs throughout the body (Wiig, Keskin andKalluri 2010, Choi, Lee andHong 2012) according to their host tropism and receptor distribution (Jang et al. 2009, Jiao et al. 2014, Op de Beeck and Eizirik 2016, Majer, McGreevy and Booth 2020, Oechslin, Moradpour and Gouttenoire 2020. ...
... Une étude pilote menée au Canada durant le mois de septembre 2014 dans sept provinces et territoires a permis de recueillir des données épidémiologiques provenant de 268 cas d'hospitalisation causés par une infection à EV-D68, parmi lesquels seulement 3 cas neurologiques ont été observés et aucun décès n'a été constaté (26). La rareté de ces cas neurologiques signifie que des études populationnelles beaucoup plus vastes devront être menées pour étudier davantage le lien possible entre l'EV-D68 et la paralysie. ...
... herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), human herpes virus type 6 (HHV-6), enteroviruses (EVs), and human paraechoviruses (HPeVs)]. [1,[19][20][21][22][23] For viral exanthems, testing often relies on several consecutive PCR assays to assess possible etiologies. The FilmArray meningitis/encephalitis (ME) panel (Biofire Diagnostics LLC, Salt Lake City, UT) is a multiplexed PCR technology that allows simultaneous detection and discrimination of viruses causing exanthems (HHV-1, HHV-2, HHV-6, VZV, EVs, and HPeVs); however, the assay is only licensed for use on cerebral spinal fluid (CSF). ...
... The VP40 monomer consists of distinct N-terminal domains (NTDs) and C-terminal domains (CTDs) that are joined through a flexible linker [22]. NTD-NTD interactions can lead to the formation of cyclic octamers that remain in the cytosol and bind RNA for regulation of EBOV replication and transcription [20,[22][23][24][25][26][27][28][29][30][31][32][33][34][35]. VP40 requires three crucial interactions to promote formation of viral particles ( Figure 1): VP40 must first form homodimers via NTD-NTD interactions for which residue L117 is crucial; such dimers accumulate at the plasma membrane due to VP40 domain rearrangement by which the CTD is flipped away from the NTD. ...
... MiRNAs play important roles during flavivirus-host interactions (Kozak et al., 2017) and it has been reported that miRNA biogenesis proteins can directly interact with flavivirus replication machinery. After infection with DENV, human cells showed a reduction in the expression levels of Drosha, Dicer, DGCR8, and AGO proteins and the silencing of these genes in virus-infected cells increases DENV replication (Kakumani et al., 2013;Casseb et al., 2016). ...