Browsing by Author "Harkins, Gordon W."
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Item Dispersal dynamics of SARS-CoV-2 lineages during the first epidemic wave in New York City(Public Library of Science, 2021) Dellicour, Simon; Hong, Samuel L.; Harkins, Gordon W.During the first phase of the COVID-19 epidemic, New York City rapidly became the epicenter of the pandemic in the United States. While molecular phylogenetic analyses have previously highlighted multiple introductions and a period of cryptic community transmission within New York City, little is known about the circulation of SARS-CoV-2 within and among its boroughs. We here perform phylogeographic investigations to gain insights into the circulation of viral lineages during the first months of the New York City outbreak. Our analyses describe the dispersal dynamics of viral lineages at the state and city levels, illustrating that peripheral samples likely correspond to distinct dispersal events originating from the main metropolitan city areas. In line with the high prevalence recorded in this area, our results highlight the relatively important role of the borough of Queens as a transmission hub associated with higher local circulation and dispersal of viral lineages toward the surrounding boroughs.Item The identification of biologically important secondary structures in disease-causing RNA viruses(University of the Western Cape, 2012) Tanov, Emil Pavlov; Harkins, Gordon W.; Christoffels, AlanViral genomes consist of either deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). The viral RNA molecules are responsible for two functions, firstly, their sequences contain the genetic code, which encodes the viral proteins, and secondly, they may form structural elements important in the regulation of the viral life-cycle. Using a host of computational and bioinformatics techniques we investigated how predicted secondary structure may influence the evolutionary dynamics of a group of single-stranded RNA viruses from the Picornaviridae family. We detected significant and marginally significant correlations between regions predicted to be structured and synonymous substitution constraints in these regions, suggesting that selection may be acting on those sites to maintain the integrity of certain structures. Additionally, coevolution analysis showed that nucleotides predicted to be base paired, tended to co-evolve with one another in a complimentary fashion in four out of the eleven species examined. Our analyses were then focused on individual structural elements within the genome-wide predicted structures. We ranked the predicted secondary structural elements according to their degree of evolutionary conservation, their associated synonymous substitution rates and the degree to which nucleotides predicted to be base paired coevolved with one another. Top ranking structures coincided with well characterized secondary structures that have been previously described in the literature. We also assessed the impact that genomic secondary structures had on the recombinational dynamics of picornavirus genomes, observing a strong tendency for recombination breakpoints to occur in non-coding regions. However, convincing evidence for the association between the distribution of predicted RNA structural elements and breakpoint clustering was not detected.Item The identification of biologically important secondary structures in disease-causing RNA viruses.(University of the Western Cape, 2012) Tanov, Emil Pavlov; Harkins, Gordon W.; Christoffels, AlanViral genomes consist of either deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). The viral RNA molecules are responsible for two functions, firstly, their sequences contain the genetic code, which encodes the viral proteins, and secondly, they may form structural elements important in the regulation of the viral life-cycle. Using a host of computational and bioinformatics techniques we investigated how predicted secondary structure may influence the evolutionary dynamics of a group of single-stranded RNA viruses from the Picornaviridae family. We detected significant and marginally significant correlations between regions predicted to be structured and synonymous substitution constraints in these regions, suggesting that selection may be acting on those sites to maintain the integrity of certain structures. Additionally, coevolution analysis showed that nucleotides predicted to be base paired, tended to co-evolve with one another in a complimentary fashion in four out of the eleven species examined. Our analyses were then focused on individual structural elements within the genome-wide predicted structures. We ranked the predicted secondary structural elements according to their degree of evolutionary conservation, their associated synonymous substitution rates and the degree to which nucleotides predicted to be base paired coevolved with one another. Top ranking structures coincided with well characterized secondary structures that have been previously described in the literature. We also assessed the impact that genomic secondary structures had on the recombinational dynamics of picornavirus genomes, observing a strong tendency for recombination breakpoints to occur in non-coding regions. However, convincing evidence for the association between the distribution of predicted RNA structural elements and breakpoint clustering was not detected.Item The molecular evolution and epidemiology of Rubella virus(University of the Western Cape, 2014) Cloete, Leendert J.; Harkins, Gordon W.Despite widespread rubella virus (RV) vaccination programs, annually RV still causes severe congenital defects in an estimated 100,000 children globally. A concerted attempt to eradicate RV is currently underway and analytical tools to monitor the global decline of the last remaining RV lineages will be useful for assessing the effectiveness of this endeavour. Importantly, RV evolves rapidly enough that much of its epidemiological information might be inferable from RV genomic sequence data. Using BEASTv1.8.0, I analysed publically available RV sequence data to estimate genome-wide and gene-specific nucleotide substitution rates, to test whether the current estimates of RV substitution rates are representative of the entire RV genome. During these investigations, I specifically accounted for possible confounders of nucleotide substitution rate estimates, such as temporally biased sampling, sporadic recombination, and natural selection favouring either increased or decreased genetic diversity (estimated by the PARRIS and FUBAR methods) at nucleotide sites within RV nucleic acid secondary structures (predicted by the NASP method). I determined that RV nucleotide substitution rates range from 1.19×10-3 substitutions/site/year (in the E1 region) to 7.52×10-4 substitutions/site/year (in the P150 region). I found that these differences between nucleotide substitution rate estimates in various RV gene regions are largely attributable to temporal sampling biases, such that datasets containing a higher proportion of recently sampled sequences will tend to have inflated estimates of mean substitution rates. Although there exists little evidence of positive selection or natural genetic recombination in RV, I revealed that RV genomes possess extensive biologically functional nucleic acid secondary structures and that purifying selection acting to maintain these structures contributes substantially to variations in estimated nucleotide substitution rates across RV genomes. Although both temporal sampling biases and purifying selection favouring the conservation of RV nucleic acid secondary structures have an appreciable impact on substitution rate estimates, I find that these biases do not preclude the use of RV sequence data to date ancestral sequences and evaluate the associated RV phylodynamics. The combination of uniformly high substitution rates across the RV genome and strong temporal signal within the available sequence data enabled me to analyse the epidemiological and demographical dynamics of this virus during these attempts to eradicate it. By implementing a generalized linear model (GLM) and symmetrical model of discretized phylogeographic spread, I was able to identify several predictive variables of geographical RV spread and detect transmission linkages between distinct geographical regions. These results suggest that, in addition to strengthened vaccination strategies, there also needs to be an increased effort to educate people about the effects of vaccination and risks of RV infection.Item The phylogeography, epidemiology and determinants of Maize streak virus dispersal across Africa and the adjacent Indian Ocean Islands(University of the Western Cape, 2015) Madzokere, Eugene T.; Harkins, Gordon W.; Martin, Darren P.Maize streak disease (MSD), caused by variants of the Maize streak virus (MSV) A strain, is the world's third and Africa’s most important maize foliar disease. Outbreaks of the disease occur frequently and in an erratic fashion across Africa and Islands in the Indian Ocean causing devastating yield losses such that the emergence, resurgence and rapid diffusion of MSV-A variants in this region presents a serious threat to maize production, farmer livelihoods and food security. To compliment current MSD management systems, a total of 689 MSV-A full genomes sampled over a 32 year period (1979-2011) from 20 countries across Africa and the adjacent Indian Ocean Islands, 286 of which were novel, were used to estimate: (i) the levels of genetic diversity using MEGA and the Sequence Demarcation Tool v1.2 (SDT); (ii) the times of occurrence and distribution of recombination using the recombination detection program (RDP v.4) and the genetic algorithm for recombination detection (GARD); (iii) selection pressure on codon positions using PARRIS and FUBAR methods implemented on the DATAMONKEY web server; (iv) reconstruct the history of spatio-temporal diffusion for MSV-A using the discrete phylogeographic models implemented in BEAST v1.8.1; (v) characterize source-sink dynamics and identify predictor variables driving MSV-A dispersal using the generalized linear models, again implemented in BEAST v1.8.1. Isolates used displayed low levels of genetic diversity (0.017 mean pairwise distance and ≥ 98% nucleotide sequence identities), and a well-structured geographical distribution where all of the 233 novel isolates clustered together with the -A1 strains. A total of 34 MSV inter-strain recombination events and 33 MSV-A intra-strain recombination events, 15 of which have not been reported in previous analyses (Owor et al., 2007, Varsani et al., 2008 and Monjane et al., 2011), were detected. The majority of intra-strain MSV-A recombination events detected were inferred to have occurred within the last six decades, the oldest and most conserved of these being events 19, 26 and 28 whereas the most recent events were 8, 16, 17, 21, 23, and 29. Intra-strain recombination events 20, 25 and 33, were widely distributed amongst East African MSV-A samples, whereas events 16, 21 and 23, occurred more frequently within West African MSV-A samples. Events 1, 4, 8, 10, 14, 17, 19, 22, 24, 25, 26, 28, and 29 were more widely distributed across East, West and Southern Africa and the adjacent Indian Ocean Islands. Whereas codon positions 12 and 19 within motif I in the coat protein transcript, and four out of the seven codon positions (147, 166, 195, 203, 242, 262, 267) in the Rep transcript (codons 195 and 203 in the Rb motif and codons 262 and 267 in site B of motif IV), evolved under strong positive selection pressure, those in the movement protein (MP) and RepA protein encoding genes evolved neutrally and under negative selection pressure respectively. Phylogeographic analyses revealed that MSV-A first emerged in Zimbabwe around 1938 (95% HPD 1904 - 1956), and its dispersal across Africa and the adjacent Indian Ocean Islands was achieved through approximately 34 migration events, 19 of which were statistically supported using Bayes factor (BF) tests. The higher than previously reported mean nucleotide substitution rate [9.922 × 10-4 (95% HPD 8.54 × 10-4 to 1.1317 × 10-3) substitutions per site per year)] for the full genome recombination-free MSV-A dataset H estimated was possibly a result of high nucleotide substitution rates being conserved among geminiviruses such as MSV as previously suggested. Persistence of MSV-A was highest in source locations that include Zimbabwe, followed by South Africa, Uganda, and Kenya. These locations were characterized by high average annual precipitation; moderately high average annual temperatures; high seasonal changes; high maize yield; high prevalence of undernourishment; low trade imports and exports; high GDP per capita; low vector control pesticide usage; high percentage forest land area; low percentage arable land; high population densities, and were in close proximity to sink locations. Dispersal of MSV-A was frequent between locations that received high average annual rainfall, had high percentage forest land area, occupied high latitudes and experienced similar climatic seasons, had high GDP per capita and had balanced maize import to export ratios, and were in close geographical proximity.Item Selection analysis identifies clusters of unusual mutational changes in omicron lineage ba.1 that likely impact spike function(Oxford University Press, 2022) Martin, Darren P.; Lytras, Spyros; Harkins, Gordon W.Among the 30 nonsynonymous nucleotide substitutions in the Omicron S-gene are 13 that have only rarely been seen in other SARS-CoV-2 sequences. These mutations cluster within three functionally important regions of the S-gene at sites that will likely impact (1) interactions between subunits of the Spike trimer and the predisposition of subunits to shift from down to up configurations, (2) interactions of Spike with ACE2 receptors, and (3) the priming of Spike for membrane fusion. We show here that, based on both the rarity of these 13 mutations in intrapatient sequencing reads and patterns of selection at the codon sites where the mutations occur in SARS-CoV-2 and related sarbecoviruses, prior to the emergence of Omicron the mutations would have been predicted to decrease the fitness of any virus within which they occurred. We further propose that the mutations in each of the three clusters therefore cooperatively interact to both mitigate their individual fitness costs, and, in combination with other mutations, adaptively alter the function of Spike. Given the evident epidemic growth advantages of Omicron overall previously known SARS-CoV-2 lineages, it is crucial to determine both how such complex and highly adaptive mutation constellations were assembled within the Omicron S-gene, and why, despite unprecedented global genomic surveillance efforts, the early stages of this assembly process went completely undetected.Item Self-maintaining or continuously refreshed? The genetic structure of Euphausia lucens populations in the Benguela upwelling ecosystem(Oxford University Press, 2013) Harkins, Gordon W.; D’amato, Maria E.; Gibbons, Mark J.Populations of Euphausia lucens over the shelf of the southern Benguela upwelling region could be self-maintaining. Alternatively, they could be continually refreshed by expatriates from the SWAtlantic that enter the system via South Atlantic Central Water in the south, before developing and then being lost through advection off Namibia. These two hypotheses are investigated here by examining geographic heterogeneity and molecular variation (cox1 and ND1) of the species across its distributional range in the Southern Hemisphere. Comparisons are made with E. vallentini, which is assumed to show panmixia associated with its circumglobal distribution between 50 and 608S. Phylogenetic analysis with mitochondrial 16S ribosomal RNA and cytochrome oxidase 1 (cox1) confirmed that E. lucens and E. vallentini represent sister taxa. Strong geographic structuring of cox1 and ND1 mtDNA genetic variation by ocean basin was recorded in E. lucens, indicating that neritic populations off South Africa are likely self-maintaining.Item Studies on the population genetics of Euphausiids: a comparison of patterns in plagic taxa displaying different distributions and life-histories(University of the Western Cape, 2006) Harkins, Gordon W.; D’Amato, M.E.; Gibbons, M.J.; Faculty of ScienceThe systematic and population genetic relationships were characterised for three ecologically related euphausiid species: Euphausia lucens, E. recurva and E. vallentini. These species have different geographical distributions and life histories. All three species have a circumpolar distribution in the Southern Hemisphere while E. recurva is also distributed in the North Pacific. DNA sequence variation was determined for three regions of mitochondrial DNA and a single nuclear gene. It was conclusively demonstrated that both E. lucens and E. vallentini represent valid taxonomic species with fixed differences observed in both the nuclear and mitochondrial genes and that the low divergences previously reported for these species with 16SrRNA and CO1 resulted from a species misidentification. It was also shown that previous attempts to date the divergence between Antarctic and Sub-Antarctic euphausiid species based on 16SrRNA distances suffer from a large overestimation due to a calculation error.Item Variant-specific introduction and dispersal dynamics of SARS-CoV-2 in New York City – from Alpha to Omicron(Public Library of Science, 2023) Dellicour, Simon; Hong, Samuel L.; Harkins, Gordon W.Since the latter part of 2020, SARS-CoV-2 evolution has been characterised by the emergence of viral variants associated with distinct biological characteristics. While the main research focus has centred on the ability of new variants to increase in frequency and impact the effective reproductive number of the virus, less attention has been placed on their relative ability to establish transmission chains and to spread through a geographic area. Here, we describe a phylogeographic approach to estimate and compare the introduction and dispersal dynamics of the main SARS-CoV-2 variants – Alpha, Iota, Delta, and Omicron – that circulated in the New York City area between 2020 and 2022. Notably, our results indicate that Delta had a lower ability to establish sustained transmission chains in the NYC area and that Omicron (BA.1) was the variant fastest to disseminate across the study area. The analytical approach presented here complements non-spatially-explicit analytical approaches that seek a better understanding of the epidemiological differences that exist among successive SARS-CoV-2 variants of concernItem The westward journey of alfalfa leaf curl virus(MDPI, 2018) Davoodi, Zohreh; Bejerman, Nicolás; Richet, Cécile; Filloux, Denis; Harkins, Gordon W.Alfalfa leaf curl virus (ALCV), which causes severe disease symptoms in alfalfa (Medicago sativa L.) and is transmitted by the widespread aphid species, Aphis craccivora Koch, has been found throughout the Mediterranean basin as well as in Iran and Argentina. Here we reconstruct the evolutionary history of ALCV and attempt to determine whether the recent discovery and widespread detection of ALCV is attributable either to past diagnostic biases or to the emergence and global spread of the virus over the past few years. One hundred and twenty ALCV complete genome sequences recovered from ten countries were analyzed and four ALCV genotypes (ALCV-A, ALCV-B, ALCV-C, and ALCV-D) were clearly distinguished. We further confirm that ALCV isolates are highly recombinogenic and that recombination has been a major determinant in the origins of the various genotypes. Collectively, the sequence data support the hypothesis that, of all the analyzed locations, ALCV likely emerged and diversified in the Middle East before spreading to the western Mediterranean basin and Argentina.