Academic literature on the topic 'Whole genome sequences (WGS)'

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Journal articles on the topic "Whole genome sequences (WGS)"

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Asif, Kinza, Denise O’Rourke, Alistair R. Legione, Pollob Shil, Marc S. Marenda, and Amir H. Noormohammadi. "Whole-genome based strain identification of fowlpox virus directly from cutaneous tissue and propagated virus." PLOS ONE 16, no. 12 (December 16, 2021): e0261122. http://dx.doi.org/10.1371/journal.pone.0261122.

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Fowlpox (FP) is an economically important viral disease of commercial poultry. The fowlpox virus (FPV) is primarily characterised by immunoblotting, restriction enzyme analysis in combination with PCR, and/or nucleotide sequencing of amplicons. Whole-genome sequencing (WGS) of FPV directly from clinical specimens prevents the risk of potential genome modifications associated with in vitro culturing of the virus. Only one study has sequenced FPV genomes directly from clinical samples using Nanopore sequencing, however, the study didn’t compare the sequences against Illumina sequencing or laboratory propagated sequences. Here, the suitability of WGS for strain identification of FPV directly from cutaneous tissue was evaluated, using a combination of Illumina and Nanopore sequencing technologies. Sequencing results were compared with the sequence obtained from FPV grown in chorioallantoic membranes (CAMs) of chicken embryos. Complete genome sequence of FPV was obtained directly from affected comb tissue using a map to reference approach. FPV sequence from cutaneous tissue was highly similar to that of the virus grown in CAMs with a nucleotide identity of 99.8%. Detailed polymorphism analysis revealed the presence of a highly comparable number of single nucleotide polymorphisms (SNPs) in the two sequences when compared to the reference genome, providing essentially the same strain identification information. Comparative genome analysis of the map to reference consensus sequences from the two genomes revealed that this field isolate had the highest nucleotide identity of 99.5% with an FPV strain from the USA (Fowlpox virus isolate, FWPV-MN00.2, MH709124) and 98.8% identity with the Australian FPV vaccine strain (FWPV-S, MW142017). Sequencing results showed that WGS directly from cutaneous tissues is not only rapid and cost-effective but also provides essentially the same strain identification information as in-vitro grown virus, thus circumventing in vitro culturing.
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Therrien, Dustin A., Kranti Konganti, Jason J. Gill, Brian W. Davis, Andrew E. Hillhouse, Jordyn Michalik, H. Russell Cross, Gary C. Smith, Thomas M. Taylor, and Penny K. Riggs. "Complete Whole Genome Sequences of Escherichia coli Surrogate Strains and Comparison of Sequence Methods with Application to the Food Industry." Microorganisms 9, no. 3 (March 16, 2021): 608. http://dx.doi.org/10.3390/microorganisms9030608.

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In 2013, the U.S. Department of Agriculture Food Safety and Inspection Service (USDA-FSIS) began transitioning to whole genome sequencing (WGS) for foodborne disease outbreak- and recall-associated isolate identification of select bacterial species. While WGS offers greater precision, certain hurdles must be overcome before widespread application within the food industry is plausible. Challenges include diversity of sequencing platform outputs and lack of standardized bioinformatics workflows for data analyses. We sequenced DNA from USDA-FSIS approved, non-pathogenic E. coli surrogates and a derivative group of rifampicin-resistant mutants (rifR) via both Oxford Nanopore MinION and Illumina MiSeq platforms to generate and annotate complete genomes. Genome sequences from each clone were assembled separately so long-read, short-read, and combined sequence assemblies could be directly compared. The combined sequence data approach provides more accurate completed genomes. The genomes from these isolates were verified to lack functional key E. coli elements commonly associated with pathogenesis. Genetic alterations known to confer rifR were also identified. As the food industry adopts WGS within its food safety programs, these data provide completed genomes for commonly used surrogate strains, with a direct comparison of sequence platforms and assembly strategies relevant to research/testing workflows applicable for both processors and regulators.
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Vestal, Grant, Steven Bruzek, Amanda Lasher, Amorce Lima, and Suzane Silbert. "Whole-Genome Sequencing for Bacterial Strain Typing Using the iSeq100 Platform." Infection Control & Hospital Epidemiology 41, S1 (October 2020): s434. http://dx.doi.org/10.1017/ice.2020.1098.

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Background: Hospital-acquired infections pose a significant threat to patient health. Laboratories are starting to consider whole-genome sequencing (WGS) as a molecular method for outbreak detection and epidemiological surveillance. The objective of this study was to assess the use of the iSeq100 platform (Illumina, San Diego, CA) for accurate sequencing and WGS-based outbreak detection using the bioMérieux EPISEQ CS, a novel cloud-based software for sequence assembly and data analysis. Methods: In total, 25 isolates, including 19 MRSA isolates and 6 ATCC strains were evaluated in this study: A. baumannii ATCC 19606, B. cepacia ATCC 25416, E. faecalis ATCC 29212, E. coli ATCC 25922, P. aeruginosa ATCC 27853 and S. aureus ATCC 25923. DNA extraction of all isolates was performed on the QIAcube (Qiagen, Hilden, Germany) using the DNEasy Ultra Clean Microbial kit extraction protocol. DNA libraries were prepared for WGS using the Nextera DNA Flex Library Prep Kit (Illumina) and sequenced at 2×150-bp on the iSeq100 according to the manufacturer’s instructions. The 19 MRSA isolates were previously characterized by the DiversiLab system (bioMérieux, France). Upon validation of the iSeq100 platform, a new outbreak analysis was performed using WGS analysis using EPISEQ CS. ATCC sequences were compared to assembled reference genomes from the NCBI GenBank to assess the accuracy of the iSeq100 platform. The FASTQ files were aligned via BowTie2 version 2.2.6 software, using default parameters, and FreeBayes version 1.1.0.46-0 was used to call homozygous single-nucleotide polymorphisms (SNPs) with a minimum coverage of 5 and an allele frequency of 0.87 using default parameters. ATCC sequences were analyzed using ResFinder version 3.2 and were compared in silico to the reference genome. Results: EPISEQ CS classified 8 MRSA isolates as unrelated and grouped 11 isolates into 2 separate clusters: cluster A (5 isolates) and cluster B (6 isolates) with similarity scores of ≥99.63% and ≥99.50%, respectively. This finding contrasted with the previous characterization by DiversiLab, which identified 3 clusters of 2, 8, and 11 isolates, respectively. The EPISEQ CS resistome data detected the mecA gene in 18 of 19 MRSA isolates. Comparative analysis of the ATCCsequences to the reference genomes showed 99.9986% concordance of SNPs and 100.00% concordance between the resistance genes present. Conclusions: The iSeq100 platform accurately sequenced the bacterial isolates and could be an affordable alternative in conjunction with EPISEQ CS for epidemiological surveillance analysis and infection prevention.Funding: NoneDisclosures: None
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Chattaway, Marie A., Ulf Schaefer, Rediat Tewolde, Timothy J. Dallman, and Claire Jenkins. "Identification of Escherichia coli and Shigella Species from Whole-Genome Sequences." Journal of Clinical Microbiology 55, no. 2 (December 14, 2016): 616–23. http://dx.doi.org/10.1128/jcm.01790-16.

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ABSTRACTEscherichia coliandShigellaspecies are closely related and genetically constitute the same species. Differentiating between these two pathogens and accurately identifying the four species ofShigellaare therefore challenging. The organism-specific bioinformatics whole-genome sequencing (WGS) typing pipelines at Public Health England are dependent on the initial identification of the bacterial species by use of a kmer-based approach. Of the 1,982Escherichia coliandShigellasp. isolates analyzed in this study, 1,957 (98.4%) had concordant results by both traditional biochemistry and serology (TB&S) and the kmer identification (ID) derived from the WGS data. Of the 25 mismatches identified, 10 were enteroinvasiveE. coliisolates that were misidentified asShigella flexneriorS. boydiiby the kmer ID, and 8 wereS. flexneriisolates misidentified by TB&S asS. boydiidue to nonfunctionalS. flexneriO antigen biosynthesis genes. Analysis of the population structure based on multilocus sequence typing (MLST) data derived from the WGS data showed that the remaining discrepant results belonged to clonal complex 288 (CC288), comprising bothS. boydiiandS. dysenteriaestrains. Mismatches between the TB&S and kmer ID results were explained by the close phylogenetic relationship between the two species and were resolved with reference to the MLST data.Shigellacan be differentiated fromE. coliand accurately identified to the species level by use of kmer comparisons and MLST. Analysis of the WGS data provided explanations for the discordant results between TB&S and WGS data, revealed the true phylogenetic relationships between different species ofShigella, and identified emerging pathoadapted lineages.
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Rabinowitz, Peter, Bar Zilberman, Yair Motro, Marilyn C. Roberts, Alex Greninger, Lior Nesher, Shalom Ben-Shimol, et al. "Whole Genome Sequence Analysis of Brucella melitensis Phylogeny and Virulence Factors." Microbiology Research 12, no. 3 (August 24, 2021): 698–710. http://dx.doi.org/10.3390/microbiolres12030050.

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Brucellosis has a wide range of clinical severity in humans that remains poorly understood. Whole genome sequencing (WGS) analysis may be able to detect variation in virulence genes. We used Brucella melitensis sequences in the NCBI Sequence Read Archive (SRA) database to assemble 248 whole genomes, and additionally, assembled 27 B. melitensis genomes from samples of human patients in Southern Israel. We searched the 275 assembled genomes for the 43 B. melitensis virulence genes in the Virulence Factors of Pathogenic Bacteria Database (VFDB) and 10 other published putative virulence genes. We explored pan-genome variation across the genomes and in a pilot analysis, explored single nucleotide polymorphism (SNP) variation among the ten putative virulence genes. More than 99% of the genomes had sequences for all Brucella melitensis virulence genes included in the VFDB. The 10 other virulence genes of interest were present across all the genomes, but three of these genes had SNP variation associated with particular Brucella melitensis genotypes. SNP variation was also seen within the Israeli genomes obtained from a small geographic region. While the Brucella genome is highly conserved, this novel and large whole genome study of Brucella demonstrates the ability of whole genome and pan-genome analysis to screen multiple genomes and identify SNP variation in both known and novel virulence genes that could be associated with differential disease virulence. Further development of whole genome techniques and linkage with clinical metadata on disease outcomes could shed light on whether such variation in the Brucella genome plays a role in pathogenesis.
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Gunasekara, A. W. A. C. W. R., L. G. T. G. Rajapaksha, and T. L. Tung. "Whole-genome sequence analysis through online web interfaces: a review." Genomics & Informatics 20, no. 1 (March 31, 2022): e3. http://dx.doi.org/10.5808/gi.20038.

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The recent development of whole-genome sequencing technologies paved the way for understanding the genomes of microorganisms. Every whole-genome sequencing (WGS) project requires a considerable cost and a massive effort to address the questions at hand. The final step of WGS is data analysis. The analysis of whole-genome sequence is dependent on highly sophisticated bioinformatics tools that the research personal have to buy. However, many laboratories and research institutions do not have the bioinformatics capabilities to analyze the genomic data and therefore, are unable to take maximum advantage of whole-genome sequencing. In this aspect, this study provides a guide for research personals on a set of bioinformatics tools available online that can be used to analyze whole-genome sequence data of bacterial genomes. The web interfaces described here have many advantages and, in most cases exempting the need for costly analysis tools and intensive computing resources.
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Atxaerandio-Landa, Aitor, Ainhoa Arrieta-Gisasola, Lorena Laorden, Joseba Bikandi, Javier Garaizar, Irati Martinez-Malaxetxebarria, and Ilargi Martinez-Ballesteros. "A Practical Bioinformatics Workflow for Routine Analysis of Bacterial WGS Data." Microorganisms 10, no. 12 (November 29, 2022): 2364. http://dx.doi.org/10.3390/microorganisms10122364.

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The use of whole-genome sequencing (WGS) for bacterial characterisation has increased substantially in the last decade. Its high throughput and decreasing cost have led to significant changes in outbreak investigations and surveillance of a wide variety of microbial pathogens. Despite the innumerable advantages of WGS, several drawbacks concerning data analysis and management, as well as a general lack of standardisation, hinder its integration in routine use. In this work, a bioinformatics workflow for (Illumina) WGS data is presented for bacterial characterisation including genome annotation, species identification, serotype prediction, antimicrobial resistance prediction, virulence-related genes and plasmid replicon detection, core-genome-based or single nucleotide polymorphism (SNP)-based phylogenetic clustering and sequence typing. Workflow was tested using a collection of 22 in-house sequences of Salmonella enterica isolates belonging to a local outbreak, coupled with a collection of 182 Salmonella genomes publicly available. No errors were reported during the execution period, and all genomes were analysed. The bioinformatics workflow can be tailored to other pathogens of interest and is freely available for academic and non-profit use as an uploadable file to the Galaxy platform.
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Ruppitsch, Werner, Ariane Pietzka, Karola Prior, Stefan Bletz, Haizpea Lasa Fernandez, Franz Allerberger, Dag Harmsen, and Alexander Mellmann. "Defining and Evaluating a Core Genome Multilocus Sequence Typing Scheme for Whole-Genome Sequence-Based Typing of Listeria monocytogenes." Journal of Clinical Microbiology 53, no. 9 (July 1, 2015): 2869–76. http://dx.doi.org/10.1128/jcm.01193-15.

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Whole-genome sequencing (WGS) has emerged today as an ultimate typing tool to characterizeListeria monocytogenesoutbreaks. However, data analysis and interlaboratory comparability of WGS data are still challenging for most public health laboratories. Therefore, we have developed and evaluated a newL. monocytogenestyping scheme based on genome-wide gene-by-gene comparisons (core genome multilocus the sequence typing [cgMLST]) to allow for a unique typing nomenclature. Initially, we determined the breadth of theL. monocytogenespopulation based on MLST data with a Bayesian approach. Based on the genome sequence data of representative isolates for the whole population, cgMLST target genes were defined and reappraised with 67L. monocytogenesisolates from two outbreaks and serotype reference strains. The Bayesian population analysis generated fiveL. monocytogenesgroups. Using all available NCBI RefSeq genomes (n= 36) and six additionally sequenced strains, all genetic groups were covered. Pairwise comparisons of these 42 genome sequences resulted in 1,701 cgMLST targets present in all 42 genomes with 100% overlap and ≥90% sequence similarity. Overall, ≥99.1% of the cgMLST targets were present in 67 outbreak and serotype reference strains, underlining the representativeness of the cgMLST scheme. Moreover, cgMLST enabled clustering of outbreak isolates with ≤10 alleles difference and unambiguous separation from unrelated outgroup isolates. In conclusion, the novel cgMLST scheme not only improves outbreak investigations but also enables, due to the availability of the automatically curated cgMLST nomenclature, interlaboratory exchange of data that are crucial, especially for rapid responses during transsectorial outbreaks.
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Chiaverini, Alexandra, Mostafa Y. Abdel-Glil, Jörg Linde, Domenico Galante, Valeria Rondinone, Antonio Fasanella, Cesare Cammà, Nicola D’Alterio, Giuliano Garofolo, and Herbert Tomaso. "Whole Genome Sequencing for Studying Bacillus anthracis from an Outbreak in the Abruzzo Region of Italy." Microorganisms 8, no. 1 (January 8, 2020): 87. http://dx.doi.org/10.3390/microorganisms8010087.

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Anthrax is a serious infectious disease caused by the gram-positive and spore-forming bacterium Bacillus anthracis. In Italy, anthrax is an endemic disease with sporadic cases each year and few outbreaks, especially in Southern Italy. However, new foci have been discovered in zones without previous history of anthrax. During summer 2016, an outbreak of anthrax caused the death of four goats in the Abruzzo region, where the disease had not been reported before. In order to investigate the outbreak, we sequenced one strain and compared it to 19 Italian B. anthracis genomes. Furthermore, we downloaded 71 whole genome sequences representing the global distribution of canonical SNP lineages and used them to verify the phylogenetic positioning. To this end, we analyzed and compared the genome sequences using canonical SNPs and the whole genome SNP-based analysis. Our results demonstrate that the outbreak strain belonged to the Trans-Eurasian (TEA) group A.Br.011/009, which is the predominant clade in Central-Southern Italy. In conclusion, the high genomic relatedness of the Italian TEA strains suggests their evolution from a common ancestor, while the spread is supposedly driven by trade as well as human and transhumance activities. Here, we demonstrated the capabilities of whole genome sequencing (WGS), which can be used as a tool for outbreak analyses and surveillance activities.
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Petronella, Nicholas, Palni Kundra, Olivia Auclair, Karine Hébert, Mary Rao, Kyle Kingsley, Katrien De Bruyne, et al. "Changes detected in the genome sequences of Escherichia coli, Listeria monocytogenes, Vibrio parahaemolyticus, and Salmonella enterica after serial subculturing." Canadian Journal of Microbiology 65, no. 11 (November 2019): 842–50. http://dx.doi.org/10.1139/cjm-2019-0235.

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Whole genome sequencing (WGS) is rapidly replacing other molecular techniques for identifying and subtyping bacterial isolates. The resolution or discrimination offered by WGS is significantly higher than that offered by other molecular techniques, and WGS readily allows infrequent differences that occur between 2 closely related strains to be found. In this investigation, WGS was used to identify the changes that occurred in the genomes of 13 strains of bacterial foodborne pathogens after 100 serial subcultures. Pure cultures of Shiga-toxin-producing Escherichia coli, Salmonella enterica, Listeria monocytogenes, and Vibrio parahaemolyticus were subcultured daily for 100 successive days. The 1st and 100th subcultures were whole-genome sequenced using short-read sequencing. Single nucleotide polymorphisms (SNPs) were identified between the 1st and final culture using 2 different approaches, and multilocus sequence typing of the whole genome was also performed to detect any changes at the allelic level. The number of observed genomic changes varied by strain, species, and the SNP caller used. This study provides insight into the genomic variation that can be detected using next-generation sequencing and analysis methods after repeated subculturing of 4 important bacterial pathogens.
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Dissertations / Theses on the topic "Whole genome sequences (WGS)"

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Haimel, Matthias. "Development of computational approaches for whole-genome sequence variation and deep phenotyping." Thesis, University of Cambridge, 2019. https://www.repository.cam.ac.uk/handle/1810/283563.

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The rare disease pulmonary arterial hypertension (PAH) results in high blood pressure in the lung caused by narrowing of lung arteries. Genes causative in PAH were discovered through family studies and very often harbour rare variants. However, the genetic cause in heritable (31%) and idiopathic (79%) PAH cases is not yet known but are speculated to be caused by rare variants. Advances in high-throughput sequencing (HTS) technologies made it possible to detect variants in 98% of the human genome. A drop in sequencing costs made it feasible to sequence 10,000 individuals including 1,250 subjects diagnosed with PAH and relatives as part of the NIHR Bioresource - Rare (BR-RD) disease study. This large cohort allows the genome-wide identification of rare variants to discover novel causative genes associated with PAH in a case-control study to advance our understanding of the underlying aetiology. In the first part of my thesis, I establish a phenotype capture system that allows research nurses to record clinical measurements and other patient related information of PAH patients recruited to the NIHR BR-RD study. The implemented extensions provide a programmatic data transfer and an automated data release pipeline for analysis ready data. The second part is dedicated to the discovery of novel disease genes in PAH. I focus on one well characterised PAH disease gene to establish variant filter strategies to enrich for rare disease causing variants. I apply these filter strategies to all known PAH disease genes and describe the phenotypic differences based on clinically relevant values. Genome-wide results from different filter strategies are tested for association with PAH. I describe the findings of the rare variant association tests and provide a detailed interrogation of two novel disease genes. The last part describes the data characteristics of variant information, available non SQL (NoSQL) implementations and evaluates the suitability and scalability of distributed compute frameworks to store and analyse population scale variation data. Based on the evaluation, I implement a variant analysis platform that incrementally merges samples, annotates variants and enables the analysis of 10,000 individuals in minutes. An incremental design for variant merging and annotation has not been described before. Using the framework, I develop a quality score to reduce technical variation and other biases. The result from the rare variant association test is compared with traditional methods.
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Gordon, Nicola. "Whole genome sequencing (WGS) as a unified platform for outbreak identification and resistance prediction in Staphylococcus aureus." Thesis, University of Oxford, 2016. https://ora.ox.ac.uk/objects/uuid:7d705b89-c5ed-4103-98fb-3f8637e88d32.

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Staphylococcus aureus continues to present challenges to modern healthcare, due to its acquisition of antimicrobial resistance factors, its ability to cause invasive infections associated with significant morbidity and mortality, and its propensity for person-to-person transmission resulting in outbreaks. For outbreak investigation, current typing methods lack resolution, and the relatively slow turnaround times may hinder effective infection control intervention. Whole-genome sequencing (WGS) is rapidly becoming faster and more affordable, offering increased resolution in a comparable timeframe. Having the entire genome means the data can also be explored for resistance and virulence testing. In this thesis, I explore the use of WGS for investigating outbreaks and for predicting antimicrobial resistance phenotype. First, I establish the genetic diversity in the nasal S. aureus population at acquisition and after long-term carriage, to determine the effect of within-host diversity on outbreak WGS interpretation. I then use 15 well characterised S. aureus outbreaks to develop an WGS approach for outbreak investigation. I test this approach by applying WGS to a further 5 outbreaks where the infection control investigation was inconclusive. Combining the epidemiological and WGS data from all 20 outbreaks, I then evaluate whether the WGS data can be used to predict the possibility of a long-term carrier involved in maintaining an outbreak, and apply this in real-time using a rapid turnaround benchtop sequencer. Finally, I explore the use of WGS for predicting antimicrobial resistance phenotype. I conclude that, for outbreaks, WGS has enhanced resolution compared to standard techniques and can give additional information to aid the outbreak investigation. For antimicrobial resistance, WGS is as sensitive and specific as routine testing methods. WGS provides a promising alternative to traditional culture and typing methods for enhancing our understanding of S. aureus and ultimately other pathogens.
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Kozma, Radoslav. "Inferring demographic history and speciation of grouse using whole genome sequences." Doctoral thesis, Uppsala universitet, Zooekologi, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-299926.

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From an ecological perspective, knowledge of demographic history is highly valuable because population size fluctuations can be matched to known climatic events, thereby revealing great insight into a species’ reaction to past climate change. This in turn enables us to predict how they might respond to future climate scenarios. Prominently, with the advent of high-throughput sequencing it is now becoming possible to assemble genomes of non-model organisms thereby providing unprecedented resolution to the study of demographic history and speciation. This thesis utilises four species of grouse (Aves, subfamily Tetraoninae) in order to explore the demographic history and speciation within this lineage; the willow grouse, red grouse, rock ptarmigan and the black grouse. I, and my co-authors, begin by reviewing the plethora of methods used to estimate contemporary effective population size (Ne) and demographic history that are available to animal conservation practitioners. We find that their underlying assumptions and necessary input data can bias in their application, and thus we provide a summary of their applicability. I then use the whole genomes of the black grouse, willow grouse and rock ptarmigan to infer their population dynamics within the last million years. I find three dominant periods that shape their demographic history: early Pleistocene cooling (3-0.9 Mya), the mid-Brunhes event (430 kya) and the last glacial period (110-10 kya). I also find strong signals of local population history – recolonization and subdivision events – affecting their demography. In the subsequent study, I explore the grouse dynamics within the last glacial period in more detail by including more distant samples and using ecological modelling to track habitat distribution changes. I further uncover strong signals of local population history, with multiple fringe populations undergoing severe bottlenecks. I also determine that future climate change is expected to drastically constrict the distribution of the studied grouse. Lastly, I use whole genome sequencing to uncover 6 highly differentiated regions, containing 7 genes, hinting at their role in adaptation and speciation in three grouse taxa. I also locate a region of low differentiation, containing the Agouti pigmentation gene, indicating its role in the grouse plumage coloration.
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Jakt, Lars Martin. "Isolation of mouse Hoxb-3 protein binding sequences : a whole genome approach /." Thesis, Hong Kong : University of Hong Kong, 1999. http://sunzi.lib.hku.hk/hkuto/record.jsp?B21185505.

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Khoo, Choon-Kiat. "Chicken genome variations and selection : from sequences to consequences." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/28934.

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Chicken is a major protein source and intensively selected for economically important traits by humans. As such, this generated a huge range of phenotypes that representing a diverse spectrum of genetic variation. Understanding the functional basis of the genetic variants that underlie these traits, however, remains a formidable endeavour particularly for complex traits. Nonetheless, molecular phenotyping of an organism from sequenced data is doable with the advances in bioinformatics analysis and unparalleled surveys of genome wide genetic variants. This provides the opportunity to gain insights into the genome architecture and assists in identifying chromosomal regions underlying selection through a “sequences to consequences” approach. Combining a whole genome re-sequencing (WGS) approach with the knowledge of selection history, this thesis aimed to study the chromosomal regions and genetic variants underlying traits of interest in various selected chicken populations. To achieve this, genetic (quantitative and population genetics), genomic and bioinformatics approaches were employed and integrated to investigate the genome wide selection signatures in a number of different lines of chicken selected for different complex traits. This includes analysing: (i) divergently selected broilers for fatness traits (Chapter 2), (ii) a closed population of layer chickens (Chapter 3), (iii) selection signatures unique to broiler or layer chickens (Chapter 4) and (iv) selection signatures in colony stimulating factor 1 (CSF1) associated with gene expression differences in broiler and layer populations (Chapter 5). Candidate genes and nucleotides underlying potential selection regions were identified, and attempts were made to further elucidate the potential interplay between genes and the biological pathways involved in regulating traits in these selected chicken lines. Incorporating integrative approaches, variants within selection signatures were annotated to provide further evidence of their functional consequences. Overall, non-coding regions were enriched in selection signatures implied that causative variants may have regulatory roles. Capitalising on the millions of genetic variants discovered from WGS, chromosomal regions subject to selection were detected using a number of population genetics statistics. In broiler chicken lines divergently selected for very low-density plasma lipoprotein (VLDL) (Chapter 2), incorporating signatures of selection helped to improve the resolution of previously mapped quantitative traits loci (QTL) intervals. This research demonstrated that the integration of the analysis of selection signatures with functional annotation of genetic variants enabled refinement and characterisation of the QTL for fatness traits. In a closed population of brown leghorn layers (Chapter 3), evidence of selection signatures was found through Tajima’s D analysis. The analysis unravelled selection signatures encoding genes involved in numerous pathways and genes having key roles such as in behaviour, including feather pecking. Combining population differentiation statistic (FST) and Tajima’s D, a number of regions subject to divergent selection between broilers and white egg layers were identified (Chapter 4). Selection signatures were found to harbour mutations involved in cellular and tissue development, including genes having important roles in growth, fatness, egg shell strength and muscle development. These regions and the overlapping genes thereby may be potentially contributing to the different phenotypic variations observed between broilers and layers. In Chapter 5, a revised gene model for colony stimulating factor 1 (CSF1) showing complex pattern of alternate transcripts was predicted from transcriptome analysis of RNA isolated from 21 different tissues. In parallel, selection signatures analysis with the FST statistic, identified selection signatures that differentiate broilers to white egg layers (3 regions) or brown egg layers to white egg layers (4 regions). All these selection signatures were located within non-coding regions, indicating potential divergent selection of CSF1 within regulatory regions. Overall, the results presented in this thesis using the “sequences to consequences” approach, link several genomic regions and genes to phenotypic variation in domesticated chicken lines. The work reported here serves as a foundation for further study to decipher the relationship between “genotype and phenotype” and its functional consequences due to selection.
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Gladstein, Ariella. "Inference of Recent Demographic History of Population Isolates Using Genome-Wide High Density SNP Arrays and Whole Genome Sequences." Thesis, The University of Arizona, 2018. http://pqdtopen.proquest.com/#viewpdf?dispub=10839026.

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In this dissertation I addressed the problem of SNP array bias when finding runs of ho- mozygosity. I demonstrated the pitfalls of using uninformed methods for finding runs of homozygosity and provide better alternatives, including a more reliable algorithm for identi- fying runs of homozygosity than the most commonly used program. I then provide a review of Ashkenazi population genetics. Next, I developed software to efficiently run millions of whole chromosome simulations, which is publicly available through GitHub, DockerHub, and on the CyVerse Discovery Environment. I applied my computational method to use Approximate Bayesian Computation to test models of Ashkenazi Jewish demographic his- tory. I found that the Ashkenazi Jews are comprised of genetically distinct subgroups from Eastern and Western Europe, as a result of massive population growth in the Eastern Ashkenazi Jews, but not in the Western Ashkenazi Jews. I further confirmed that the Ashkenazi Jews do not primarily originate from Khazaria. Finally, I created a correction for SNP array ascertainment bias in the median and total length of runs of homozygosity, and applied this correction to world-wide human populations. However, I found that ascertainment bias plays a minor role compared to SNP array bias in human populations.

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Alosaimi, Shatha Mobarak. "Leveraging Whole Genome Sequences to Compare Mutational Mechanism and Identify Medically Relevant Variation in African versus Non-African Descend Populations." Master's thesis, Faculty of Health Sciences, 2020. http://hdl.handle.net/11427/32191.

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Whole-Genome Sequencing (WGS) is ushering a new era in healthcare and research in identifying genetic variation in all populations. However, the African populations are still under-represented. Since African populations are being the most genetically diverse with high heterogeneity rate, we need to benchmark the Whole Genome Sequence (WGS) analysis pipeline to ensure reliable mutation detection. Therefore, it is essential to ensure that all steps of WGS downstream analysis are accurate, mainly the variant calling (VC). Current VC tools may produce falsepositive/negative results; such result may produce misleading conclusions in prioritisation of mutation, clinical relevancy and actionability of genes. With such many VC tools, two questions have arisen. Firstly, which tool has a high rate of sensitivity and precision in low either high coverage African sequences, given they have high genetic diversity and heterogeneity? Secondly, does the improvement of the VC result will advance the accuracy of detecting mutation and incidental finding (actionable genes) in African populations? In this project, a total of 100 DNA sequence samples was simulated (of which every 50 samples mimicked the genetics background of African and European, respectively) at different coverage (high and low). In particular, the sensitivity to discover polymorphisms was done by nine different VC tools. These tools were assessed in term of false positive/negative call rate given the simulated golden variants. Combining our result on sensitivity and positive predictive value (PPV). Lofreq performs best in African population data (sens=0.85, PPV=0.983, F-score=0.91) on high/low coverage data; as a result, we chose Lofreq to perform variant calling, and Gene-based annotation is performed to conduct in-sillico predication of mutation on publicly available data (the African Genome Variation and 1000 Genome Project). In doing so, we have leveraged WGS to examine and validate four of burden diseases in the African content, such as communicable diseases: HIV/AIDS, Malaria, Tuberculosis (TB), and Non-communicable diseases: such as Sickle cell disease, these diseases have uniquely shaped ethnic-specific and continental genomics variation and therefore provides unprecedented opportunities to map disease genes across the African continent. Moreover, the current actionable gene recommended by The American College of Medical Genetics and Genomics (ACMG) in the African population and update on additional African-specific actionable genes. Our result suggests African and African diaspora ethnic groups, particularly Bantu and Khoesan ethnics have gene diversity, high proportion of derived allele at low minor allele frequency (0.0 − 01) and the highest proportion of pathogenic variants within HIV, TB, Malaria, Sickle-Cell disease, while non-African ethnic groups including Latin America, Afro-Asiatic European related ethnic groups have high proportion of pathogenic variants within current actionable gene list. Overall, given the observed highest genetic diversity found in African ethnics and African diaspora related ethnics at these four Africa burden diseases and current actionable gene associated, our results support (1) the use of personalised medicine as beneficial to both African continent and worldwide; (2) a recommendation for African-specific actionable list of genes to further improve African and diaspora healthcare.
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Wiredu, Boakye Dominic. "Life in the nucleus : the genomic basis of energy exploitation by intranuclear Microsporidia." Thesis, University of Exeter, 2016. http://hdl.handle.net/10871/26108.

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The Microsporidia are obligate intracellular parasites that have jettisoned oxidation phosphorylative capabilities during their early evolutionary history and so rely on ATP import from their host and glycolysis for their energy needs. Some species form tight associations with the host’s mitochondria and this is thought to facilitate ATP sequestration by the developing intracellular microsporidian. The human parasite, Enterocytozoon bieneusi has however lost glycolytic capabilities and may rely entirely on ATP import from its host for energy. E. bieneusi belongs to the Enterocytozoonidae microsporidian family and recent rDNA-based phylogenetic studies have suggested it has close evolutionary ties with Enterospora canceri, a crab-infecting intranuclear parasite. Such a close evolutionary relationship implied that glycolysis might also be absent in the intranuclear parasite raising questions as to how this parasite obtains energy from its unusual niche that is physically walled off from the host mitochondria, the main source of ATP in the host cell. In this study, draft genomes of four species of the Enterocytozoonidae namely, Ent. canceri, E. hepatopenaei, Hepatospora eriocheir and Hepatospora eriocheir canceri and one non-Enterocytozoonidae species, Thelohania sp. were assembled and annotated (The genome assembly of Hepatospora eriocheir was provided by Dr. Bryony Williams). Phylogenomics performed with this and publicly available genomic data confirmed the close evolutionary ties between Ent. canceri and E. bieneusi. Comparative genomic analyses also revealed that glycolysis is indeed lost in all members of the Enterocytozoonidae family sequenced in this study, hinting to the relaxation of evolutionary pressures to maintain this pathway at the base of this microsporidian family. Despite this absence, the hexokinase gene was retained in all aglycolytic genomes analysed, and that of Ent. canceri was fused to a PTPA gene. Functional assays and yeast complementation assays suggest that this chimera is able to recognise glucose as a substrate but the heterologously expressed homolog of H. eriocheir cannot. Finally, phylogenomics have been used here to demonstrate that despite the morphological differences between three Hepatospora-like organisms parasitizing different crab hosts, they are the same species. This finding adds more weight to current evidence suggesting that morphology is not an ideal marker for taxonomical classification in the Microsporidia.
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Jäger, Sarah Christina [Verfasser]. "Hybrid Assembly of Whole Genome Shotgun Sequences of Two Sugar Beet (Beta vulgaris L.) Translocation Lines Carrying the Beet Cyst Nematode Resistance Gene Hs1-2 and Functional Analysis of Candidate Genes / Sarah Christina Jäger." Kiel : Universitätsbibliothek Kiel, 2013. http://d-nb.info/1054661898/34.

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Font, Porterias Neus 1994. "Genomic insights into an underrepresented population : the Romani." Doctoral thesis, TDX (Tesis Doctorals en Xarxa), 2022. http://hdl.handle.net/10803/673740.

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La població romaní és el grup ètnic minoritari transnacional més gran d'Europa. Tenen un origen sud asiàtic i, durant la seva diàspora cap a Europa, van experimentar múltiples efectes fundadors i una barreja genètica extensa. En aquesta tesi, l’anàlisi de marcadors genètics autosòmics mostren que tots els romaní comparteixen un component genètic comú del sud d’Àsia, de l’Orient Mitjà i dels Balcans, mentre que els grups romaní de la Península Ibèrica i del Bàltic han tingut una barreja addicional amb les altres poblacions de l'entorn. Després de caracteritzar el seu panorama genètic, l’estudi d’exomes complets de romanís espanyols suggereix que el flux de gens no romanís ha contrarestat l’augment de mutacions deletèries causada pels efectes fundadors. A més, les variants clínicament rellevants es troben tant a haplotips europeus com sud-asiàtics consistent amb l’extensa barreja genètica. Aquest treball representa un pas endavant per entendre de manera exhaustiva la història demogràfica romaní i posa de manifest la necessitat d’estudiar poblacions històricament excloses per poder descriure la variació humana en la seva totalitat.
The Romani people are the largest transnational minority ethnic group in Europe. They have a South Asian origin and during their diaspora to Europe, they experienced multiple founder effects and gene flow events. In this thesis, the analysis of genome-wide array data shows that Romani groups share a common South Asian, Middle Eastern and Balkan ancestry, while Iberian and Baltic groups experienced additional admixture with the surrounding non-Roma European populations. After characterising their genetic landscape, the study of whole-exome sequences from Spanish Romani suggests that non-Roma gene flow has counteracted the increase in mutational load caused by the founder effects. In addition, clinically relevant variants are traced back to both European and South Asian ancestral haplotypes consistent with the extensive gene flow. Thus, the present work represents a step forward to comprehensively depict the Romani demographic history and emphasises the need to study other underrepresented and historically excluded populations to fully capture human variation.
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Books on the topic "Whole genome sequences (WGS)"

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Crawford, Michael, and Rohina C. Rubicz. Molecular Genetic Evidence from Contemporary Populations for the Origins of Native North Americans. Edited by Max Friesen and Owen Mason. Oxford University Press, 2016. http://dx.doi.org/10.1093/oxfordhb/9780199766956.013.4.

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An overview of the current molecular genetic evidence for the origins of North American populations is presented, including specific examples from the authors’ work with the Aleutian Island inhabitants. Shared mitochondrial DNA and Y-chromosome DNA markers among Siberians and Native Americans point to a Pleistocene migration from Siberia into the Americas via Beringia. There was likely a later migration from Siberia to Alaska, based on the analysis of whole-genome sequence data from a Greenland Paleoeskimo that clusters this individual with Siberian populations. Coalescence date estimates for Native American mitochondrial DNA and Y-chromosome DNA haplogroups indicate that there was a population expansion approximately 15,000–18,000 that was associated with a pre-Clovis settlement of the Americas and coastal migration, and then a later expansion of circum-Arctic populations. Settlement of the Aleutian Archipelago took place via east-to-west migration of Aleut kin groups, accompanied by a clinal loss in mitochondrial DNA haplotype diversity.
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Muir, Stephanie. Studying City of God. Liverpool University Press, 2008. http://dx.doi.org/10.3828/liverpool/9781903663592.001.0001.

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A leading example of a resurgent Latin American cinema — ‘la buena onda’ — in the early twenty-first century, City of God was a huge international popular and critical success. A combination of intoxicating, Hollywood-style genre film-making and hard-hitting, social-realist subject matter, it was hailed as a masterpiece at Cannes in 2002 and seen by over 3 million people in Brazil, including the Brazilian cabinet. This book considers: The historical and industrial context of City of God — a brief history of Latin American cinema is followed by a more detailed account of film-making in Brazil — from light-hearted travelogues to Cinema Novo and after — all in the context of increasing globalisation; Narrative and Genre — how the film uses the components of narrative in a complex way, experimentally manipulating time while using traditional genre conventions that are highly recognisable to mainstream audiences; Film language — the formal elements of the film are dissected through a detailed illustrated analysis of the kinetic, scene setting opening sequence; Audience responses — from establishment critical reaction to fan-based internet sites and student feedback; Representation and Ideology — just how ‘authentic’ can a film such as City of God hope to be? Does its style overwhelm its subject matter?
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Ayala, Francisco J., and Camilo J. Cela-Conde. Neanderthals and modern humans. Oxford University Press, 2017. http://dx.doi.org/10.1093/oso/9780198739906.003.0011.

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This chapter deals with the similarities and differences between Homo neanderthalensis and Homo sapiens, by considering genetic, brain, and cognitive evidence. The genetic differentiation emerges from fossil genetic evidence obtained first from mtDNA and later from nuclear DNA. With high throughput whole genome sequencing, sequences have been obtained from the Denisova Cave (Siberia) fossils. Nuclear DNA of a third species (“Denisovans”) has been obtained from the same cave and used to define the phylogenetic relationships among the three species during the Upper Palaeolithic. Archaeological comparisons make it possible to advance a four-mode model of the evolution of symbolism. Neanderthals and modern humans would share a “modern mind” as defined up to Symbolic Mode 3. Whether the Neanderthals reached symbolic Mode 4 remains unsettled.
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Bilański, Piotr. Trypodendron laeve Eggers w Polsce na tle wybranych aspektów morfologicznych i genetycznych drwalników (Trypodendron spp., Coleoptera, Curculionidae, Scolytinae). Publishing House of the University of Agriculture in Krakow, 2019. http://dx.doi.org/10.15576/978-83-66602-38-0.

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In Poland, there are 4 species of the liypodendron genus: T lineaium Oliv., T domestkum L., T signature Fakir. and 7: laeve Egg. Trypodendron laeve is the leastknown of this group. Many factors had influence on the state of research on this species, including taxonomic aspects. Taking into account the unsatisfactory state of knowledge regarding the prevalence of T iaeve in Poland, as well as scarce information on the morphology of this species, research was undertaken to I) document the presence, including new sites, of T laeve in Poland and define, if possible, the habitat and trophic conditions that may affect its occurrence, as well as II) determinate suitability of biometric and genetic methods for correct identification of t laeve against the background of other ambrosia beetle species. Research on the occurrence of T laeve in Poland, was carried out on 143 areas located throughout the country, representing various environmental conditions, primarily such as species composition of tree stands, terrain, altitude (from 16 to 929 meters above sea level) and their location in relation to zoogeographic regions. The research material was obtained mainly using various types of traps for catching ambrosia beetles baited with pheromone. Only in a few cases when attacking the wood of trees, the imagines of ambrosia beetles were obtained without luring agents. The research was conducted in 2007-2016. From the insect individuals identified on the grounds of morphological traits as T lineatum, T laeve, T domesticum and T signatum, originating from selected locations in Poland, 3-11 specimens were collected, for which genetic analyses were performed based on the COI gene fragments obtained by PCR. The research included tests for following paramcter: s sequence similarity, phylogenetic, evolutionary divergence and genetic. structure. As a result of research on the occurrence of ambrosia beetles in Poland, a total of 44207 individuals belonging to four species were collected: T lineatutn, 7: laeve, T domesticum and T signatum, whose share was respectively: 49.2%, 31.4%; 19.1% and 0.3%. In Poland, 1: laeve's imagines were found in 124 out of 143 examined sites. The presence of L reeve has been documented for the first time in 14 zoogeographic regions. This species was commonly found on study areas located from 118 to 929 m above sea level. In Poland the tree species attacked by T Mate include Pinus sylvestris L. and Picea abies (L) H. Karst. In Poland, T laeve as a host plant prefers sylvestris and reaches the highest population densities in the stands of this species. The work presents the exact morphological characteristics of T laeve and indicates the most important features that distinguish it from the other Trypodendrun spp. occurring in Poland. It has also been shown that the best results in the determination of species of the liypodendron genus, regardless of their sex, can be obtained using phylogenetic analysis based on a fragment of the COI gene.
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Book chapters on the topic "Whole genome sequences (WGS)"

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Stessl, Beatrix, Martin Wagner, and Werner Ruppitsch. "Multilocus Sequence Typing (MLST) and Whole Genome Sequencing (WGS) of Listeria monocytogenes and Listeria innocua." In Listeria Monocytogenes, 89–103. New York, NY: Springer US, 2020. http://dx.doi.org/10.1007/978-1-0716-0982-8_7.

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Arnemann, J. "Whole-Genome Sequenzierung (WGS)." In Springer Reference Medizin, 2509. Berlin, Heidelberg: Springer Berlin Heidelberg, 2019. http://dx.doi.org/10.1007/978-3-662-48986-4_3632.

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Arnemann, J. "Whole-Genome Sequenzierung (WGS)." In Lexikon der Medizinischen Laboratoriumsdiagnostik, 1. Berlin, Heidelberg: Springer Berlin Heidelberg, 2018. http://dx.doi.org/10.1007/978-3-662-49054-9_3632-1.

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Bharadwaj, Shiv, Vivek Dhar Dwivedi, and Nikhil Kirtipal. "Application of Whole Genome Sequencing (WGS) Approach Against Identification of Foodborne Bacteria." In Microbial Genomics in Sustainable Agroecosystems, 131–48. Singapore: Springer Singapore, 2019. http://dx.doi.org/10.1007/978-981-13-8739-5_7.

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Shirasawa, Kenta, and Hiroyasu Kitashiba. "Genetic Maps and Whole Genome Sequences of Radish." In Compendium of Plant Genomes, 31–42. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-59253-4_3.

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Gupta, Manoj Kumar, Ravindra Donde, S. Sabarinathan, Gayatri Gouda, Goutam Kumar Dash, Pallabi Pati, Sushil Kumar Rathore, et al. "Microsatellite Markers from Whole Genome and Transcriptomic Sequences." In Bioinformatics in Rice Research, 387–412. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-16-3993-7_18.

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Hahn, Georg, Sharon Marie Lutz, Julian Hecker, Dmitry Prokopenko, and Christoph Lange. "Local and Global Stratification Analysis in Whole Genome Sequencing (WGS) Studies Using LocStra." In Computational Advances in Bio and Medical Sciences, 159–70. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-46165-2_13.

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Rodriguez-R, Luis M., Ramon Rosselló-Móra, and Konstantinos T. Konstantinidis. "Identification and Classification of Prokaryotes using whole-genome sequences." In Trends in the systematics of bacteria and fungi, 217–30. Wallingford: CABI, 2021. http://dx.doi.org/10.1079/9781789244984.0217.

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Abstract This book chapter attempts to summarize the major findings from genome-based taxonomic studies in the past two decades, and briefly describe the major genome-based approaches currently available for species identification and classification with special focus on the 'uncultivated majority' and associated limitations, as well as outlines future directions towards a truly genome-based taxonomy for prokaryotes that will equally encompass cultured and uncultivated taxa. Importantly, the need for a system to catalogue uncultivated taxa is very urgent, because the genomes and ecological/functional data that are becoming available are already overwhelming, and alphanumeric identifiers and synonyms are creating confusion of Babylonian dimensions.
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Baroncelli, Riccardo, and Giovanni Cafà. "Genomic sequences for fungi." In Trends in the systematics of bacteria and fungi, 231–54. Wallingford: CABI, 2021. http://dx.doi.org/10.1079/9781789244984.0231.

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Abstract This chapter aims to give an overview of the basic knowledge, understanding and perspectives in fungal genomics. It is likely that fungal genome sequencing will soon become simpler and cheaper, and allow most research laboratories to undertake in-house, whole genome sequencing on a regular basis, as sequencers will be accessible to most laboratories. Nonetheless, most of the innovation in the next decade will be driven by theories in innovative perspectives and fields of investigation, rather than in novel technical approaches.
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Riojas, Marco A., Andrew M. Frank, Samuel R. Greenfield, Stephen P. King, Conor J. Meehan, Michael Strong, Alice R. Wattam, and Manzour Hernando Hazbón. "Identification and Characterization of Mycobacterial Species Using Whole-Genome Sequences." In Methods in Molecular Biology, 399–457. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1460-0_19.

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Conference papers on the topic "Whole genome sequences (WGS)"

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Lange, I., C. Stein, J. Rödel, A. Knabe, F. Kipp, H. Proquitté, and K. Dawczynski. "Analyse eines S. aureus-Clusters in der Neonatologie mittels Whole Genome Sequencing (WGS)." In 29. Deutscher Kongress für Perinatale Medizin. Deutsche Gesellschaft für Perinatale Medizin (DGPM) – „Hinterm Horizont geht's weiter, zusammen sind wir stark“. Georg Thieme Verlag KG, 2019. http://dx.doi.org/10.1055/s-0039-3401202.

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Guro, P., V. Safronova, A. Sazanova, I. Kuznetsova, A. Belimov, V. Yakubov, E. Chirak, A. Afonin, E. Andronov, and I. Tikhonovich. "Rhizobial microsymbionts of the narrowly endemic Oxytropis species growing in Kamchatka possess a set of genes that are associated with T3SS and T6SS secretion systems and can affect the development of symbiosis." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.099.

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A collection of rhizobial strains isolated from root nodules of the narrowly endemic legume species Oxytropis erecta, O. anadyrensis, O. kamtschatica and O. pumilio growing on the Kamchatka Peninsula (Russian Federation) was obtained. Analysis of the 16S rRNA gene sequence showed a significant diversity of isolates belonging to the families Rhizobiaceae (Rhizobium), Phyllobacteriaceae (Mesorhizobium, Phyllobacterium) and Bradyrhizobiaceae (Bosea, Tardiphaga). Pairs of taxonomically different strains in various combinations were isolated from some nodules of Oxytropis plants. Plant nodulation assays showed that only strains belonging to the genus Mesorhizobium (M. jarvisii, M. loti and M. huakuii) could form nitrogen-fixing nodules. The nitrogen-fixing activity of the strains was more associated with the host plant than with the species of strains. The whole genome sequences analysis showed that the strains M. loti 582 and M. huakuii 583 possessed symbiotic genes necessary for the formation of effective symbiosis and grouped into Sym-clusters. In contrast, the strain T. robiniae 581 had only a reduced number of fix genes, while the strains Phyllobacterium sp. 628 and R. lusitanum 629 possesed only individual symbiotic genes, which obviously did not participate in the formation of nodules. It was also stated that the strains M. loti 582 and M. huakuii 583 had a significantly larger set of genes related to the secretion systems T3SS and T6SS that can affect the host specificity of strains, compared with 6 commercial strains used as reference. These two strains formed nodules of two types (typical elongated and atypical rounded) on Oxytropis plants. We suggest that a possible cause of the observed phenomenon is the availability of different nodulation strategies in these strains (dependent and independent of Nod-factors). Thus, as a result of studying the collection of strains isolated from the narrow endemic species of Kamchatka Oxytropis, interesting objects were selected to study the functions of the T3SS and T6SS genes, and their role in the development of rhizobia-legume symbiosis. The prospects of using strains with gene systems for both symbiotic and non-symbiotic nodulation to enhance the efficiency of plant-microbe interactions by expanding the host specificity and increasing the efficiency of nodulation are discussed.
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Ganapathiraju, M., D. Weisser, R. Rosenfeld, J. Carbonell, R. Reddy, and J. Klein-Seetharaman. "Comparative n-gram analysis of whole-genome protein sequences." In the second international conference. Morristown, NJ, USA: Association for Computational Linguistics, 2002. http://dx.doi.org/10.3115/1289189.1289259.

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Kaptelova, V. V., A. S. Speranskaya, A. E. Samoilov, A. V. Valdokhina, V. P. Bulanenko, E. V. Korneenko, O. Y. Shipulina, and V. G. Akimkin. "MUTATIONS IN THE GENOMES OF SARS-COV-2 FROM CLINICAL SAMPLES OBTAINED IN LATE MARCH-EARLY APRIL FROM PATIENTS IN MOSCOW." In Molecular Diagnostics and Biosafety. Federal Budget Institute of Science 'Central Research Institute for Epidemiology', 2020. http://dx.doi.org/10.36233/978-5-9900432-9-9-147.

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Many papers suggested that D614G mutation in the viral spike (S) protein SARS-CoV-2 can influence the ability of virus transmission. In recent work [1], it was shown D614G influences the rate of disease transmission only in combination with the P323L mutation in the viral polymerase. We have sequenced 28 full genomes of SARS-CoV-2, obtained from clinical material from patients of different ages. The analyzed isolates belong to clades B.1 (GH) and B1.1 (GR). Combinations of mutations P323L and D614G were found in all genomes. These differences can be explained by sampling: the samples for the sequencing of the whole genome were selected with high viral load, it can be related to the rate of viral replication in intra-host. That, in turn, can be dependent on the presence of P323L/D614G mutations in the virus genome.
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O'hara, Patrick J., Frank A. Grant, A. Betty, J. Haldmen, and Mark J. Murray. "Structure of the Human Factor VII Gene." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643786.

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Factor VII is a member of a family of vitamin K-dependent, gamma-carboxylated plasma protein which includes factor IX, factor X, protein C, protein S and prothrombin. Activated factor VII (factor Vila) is a plasma serine protease which participates in a cascade of reactions leading to the coagulation of blood. Two overlapping genomic clones containing sequences encoding human factor VII were isolated and characterized. The complete sequence of the gene was determined and found to span 12.8 kilobases. The mRNA for factor VII as demonstrated by cDNA cloning is polyadenylated at multiple sites but contains only one AAUAAA poly-A signal sequence. The mRNA can undergo alternative splicing forming one transcript containing eight segments as exons and another with an additional exon which encodes a larger pre-pro leader sequence. The portion of the pre-pro leader coded for by the additional exon has no known counterpart in the other vitamin K-dependent proteins. The positions of the introns with respect to the amino acid sequence encoded by the eight essential exons of factor VII are the same as those present in factor IX, factor X, protein C and the first three exons of prothrombin. These exons code for domains generally conserved among members of this gene family, including a pre-pro leader (the essential exon la and alternative exon lb), a gamma-carboxylated domain (exons 2 and 3) a growth factor domain (exons 4 and 5) an activation region (exon 6) and a serine protease (exon 8). The corresponding introns in these genes are dissimilar with respect to size and sequence, with the exception of the third intron in factor VII and protein C. Four introns and a portion of exon 8 in factor VII contain regions made up of tandem repeats of oligonucleotide monomer elements. More than a quarter of the intron sequences and more than a third of the 3' untranslated portion of the mRNA transcript consist of these minisatellite tandem repeats. This type of structure is responsible for polymorphisms due to allelic variation in repeat copy number in other areas of the human genome. Tandem repeats can evolve as a result of random crossover in DNA whose sequence is not maintained by selection. This suggests that much of the sequence information present in the introns and untranslated portion of the message is dispensable.
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Wei Zhang, Erliang Zeng, Scott J. Emrich, Joshua Livermore, Dan Liu, and Stuart E. Jones. "Predicting bacterial functional traits from whole genome sequences using random forest." In 2013 IEEE 3rd International Conference on Computational Advances in Bio and Medical Sciences (ICCABS). IEEE, 2013. http://dx.doi.org/10.1109/iccabs.2013.6629212.

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Martins, Wellington S., Juan del Cuvillo, Wenwu Cui, and Guang R. Gao. "Whole Genome Alignment using a Multithreaded Parallel Implementation." In Simpósio de Arquitetura de Computadores e Processamento de Alto Desempenho. Sociedade Brasileira de Computação, 2001. http://dx.doi.org/10.5753/sbac-pad.2001.22185.

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Alignment of long DNA sequences is a challenging task due to its high demands for computational power and memory. We have developed a multithreaded parallel implementation of a sequence alignment algorithm that is able to align whole genomes with reliable output and reasonable cost. The implementation is based on a fine-grain multithreaded execution model, the EARTH model, which effectively tolerates latency through the overlapping of computation and communication. Human and mice mitochondrial genomes, human and Drosophila mitochondrial genomes are aligned respectively to demonstrate that the implementation can be used to align both closely related as well as less similar genomes. Results from Mycoplasma genitalium and Mycoplasma pneumoniae genomes, which are much larger than the tested mitochondrial genomes, are also presented. From the output, the homologous regions can be easily detected. This tool should facilitate alignment of syntenic regions, strain to strain comparisons, identification of regulatory elements and evolutionary comparisons as well.
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Pavel, Ionut, Ina Iuliana Macovei, Simona Péter, Daniela Diculencu, Florin Rusu-Cordunean, and Bogdan Dragos Grigoriu. "Detection of first and second line drug resistance mutations from multi drug resistantmycobacterium tuberculosisstrains by Ion Torrent whole genome sequencing (WGS)." In ERS International Congress 2016 abstracts. European Respiratory Society, 2016. http://dx.doi.org/10.1183/13993003.congress-2016.pa1910.

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Qian, Jiating, Mengjiao Li, Yifan Feng, Wenjuan Li, and Jie Li. "Genetic Epidemiology of Porcine Transmissible Gastroenteritis Virus Based on Whole Genome and S Gene Sequences." In 2021 IEEE 9th International Conference on Bioinformatics and Computational Biology (ICBCB). IEEE, 2021. http://dx.doi.org/10.1109/icbcb52223.2021.9459208.

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Li, X., S. Krishnamurthy, S. Kumar, S. Reddy, W. Woodward, J. Reuben, C. Hatzis, NT Ueno, M. Gerstein, and L. Pusztai. "Abstract P1-05-01: Landscape of somatic mutations in inflammatory breast cancer whole-genome sequences." In Abstracts: 2016 San Antonio Breast Cancer Symposium; December 6-10, 2016; San Antonio, Texas. American Association for Cancer Research, 2017. http://dx.doi.org/10.1158/1538-7445.sabcs16-p1-05-01.

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Reports on the topic "Whole genome sequences (WGS)"

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Gur, Amit, Edward Buckler, Joseph Burger, Yaakov Tadmor, and Iftach Klapp. Characterization of genetic variation and yield heterosis in Cucumis melo. United States Department of Agriculture, January 2016. http://dx.doi.org/10.32747/2016.7600047.bard.

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Project objectives: 1) Characterization of variation for yield heterosis in melon using Half-Diallele (HDA) design. 2) Development and implementation of image-based yield phenotyping in melon. 3) Characterization of genetic, epigenetic and transcriptional variation across 25 founder lines and selected hybrids. The epigentic part of this objective was modified during the course of the project: instead of characterization of chromatin structure in a single melon line through genome-wide mapping of nucleosomes using MNase-seq approach, we took advantage of rapid advancements in single-molecule sequencing and shifted the focus to Nanoporelong-read sequencing of all 25 founder lines. This analysis provides invaluable information on genome-wide structural variation across our diversity 4) Integrated analyses and development of prediction models Agricultural heterosis relates to hybrids that outperform their inbred parents for yield. First generation (F1) hybrids are produced in many crop species and it is estimated that heterosis increases yield by 15-30% globally. Melon (Cucumismelo) is an economically important species of The Cucurbitaceae family and is among the most important fleshy fruits for fresh consumption Worldwide. The major goal of this project was to explore the patterns and magnitude of yield heterosis in melon and link it to whole genome sequence variation. A core subset of 25 diverse lines was selected from the Newe-Yaar melon diversity panel for whole-genome re-sequencing (WGS) and test-crosses, to produce structured half-diallele design of 300 F1 hybrids (MelHDA25). Yield variation was measured in replicated yield trials at the whole-plant and at the rootstock levels (through a common-scion grafted experiments), across the F1s and parental lines. As part of this project we also developed an algorithmic pipeline for detection and yield estimation of melons from aerial-images, towards future implementation of such high throughput, cost-effective method for remote yield evaluation in open-field melons. We found extensive, highly heritable root-derived yield variation across the diallele population that was characterized by prominent best-parent heterosis (BPH), where hybrids rootstocks outperformed their parents by 38% and 56 % under optimal irrigation and drought- stress, respectively. Through integration of the genotypic data (~4,000,000 SNPs) and yield analyses we show that root-derived hybrids yield is independent of parental genetic distance. However, we mapped novel root-derived yield QTLs through genome-wide association (GWA) analysis and a multi-QTLs model explained more than 45% of the hybrids yield variation, providing a potential route for marker-assisted hybrid rootstock breeding. Four selected hybrid rootstocks are further studied under multiple scion varieties and their validated positive effect on yield performance is now leading to ongoing evaluation of their commercial potential. On the genomic level, this project resulted in 3 layers of data: 1) whole-genome short-read Illumina sequencing (30X) of the 25 founder lines provided us with 25 genome alignments and high-density melon HapMap that is already shown to be an effective resource for QTL annotation and candidate gene analysis in melon. 2) fast advancements in long-read single-molecule sequencing allowed us to shift focus towards this technology and generate ~50X Nanoporesequencing of the 25 founders which in combination with the short-read data now enable de novo assembly of the 25 genomes that will soon lead to construction of the first melon pan-genome. 3) Transcriptomic (3' RNA-Seq) analysis of several selected hybrids and their parents provide preliminary information on differentially expressed genes that can be further used to explain the root-derived yield variation. Taken together, this project expanded our view on yield heterosis in melon with novel specific insights on root-derived yield heterosis. To our knowledge, thus far this is the largest systematic genetic analysis of rootstock effects on yield heterosis in cucurbits or any other crop plant, and our results are now translated into potential breeding applications. The genomic resources that were developed as part of this project are putting melon in the forefront of genomic research and will continue to be useful tool for the cucurbits community in years to come.
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Sherman, Amir, Rebecca Grumet, Ron Ophir, Nurit Katzir, and Yiqun Weng. Whole genome approach for genetic analysis in cucumber: Fruit size as a test case. United States Department of Agriculture, December 2013. http://dx.doi.org/10.32747/2013.7594399.bard.

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The Cucurbitaceae family includes a broad array of economically and nutritionally important crop species that are consumed as vegetables, staple starches and desserts. Fruit of these species, and types within species, exhibit extensive diversity as evidenced by variation in size, shape, color, flavor, and others. Fruit size and shape are critical quality determinants that delineate uses and market classes and are key traits under selection in breeding programs. However, the underlying genetic bases for variation in fruit size remain to be determined. A few species the Cucurbitaceae family were sequenced during the time of this project (cucumber was already sequenced when the project started watermelon and melon sequence became available during the project) but functional genomic tools are still missing. This research program had three major goals: 1. Develop whole genome cucumber and melon SNP arrays. 2. Develop and characterize cucumber populations segregating for fruit size. 3. Combine genomic tools, segregating populations, and phenotypic characterization to identify loci associated with fruit size. As suggested by the reviewers the work concentrated mostly in cucumber and not both in cucumber and melon. In order to develop a SNP (single nucleotide polymorphism) array for cucumber, available and newly generated sequence from two cucumber cultivars with extreme differences in shape and size, pickling GY14 and Chinese long 9930, were analyzed for variation (SNPs). A large set of high quality SNPs was discovered between the two parents of the RILs population (GY14 and 9930) and used to design a custom SNP array with 35000 SNPs using Agilent technology. The array was validated using 9930, Gy14 and F1 progeny of the two parents. Several mapping populations were developed for linkage mapping of quantitative trait loci (QTL) for fruit size These includes 145 F3 families and 150 recombinant inbred line (RILs F7 or F8 (Gy14 X 9930) and third population contained 450 F2 plants from a cross between Gy14 and a wild plant from India. The main population that was used in this study is the RILs population of Gy14 X 9930. Phenotypic and morphological analyses of 9930, Gy14, and their segregating F2 and RIL progeny indicated that several, likely independent, factors influence cucumber fruit size and shape, including factors that act both pre-anthesis and post-pollination. These include: amount, rate, duration, and plane of cell division pre- and post-anthesis and orientation of cell expansion. Analysis of F2 and RIL progeny indicated that factors influencing fruit length were largely determined pre-anthesis, while fruit diameter was more strongly influenced by environment and growth factors post-anthesis. These results suggest involvement of multiple genetically segregating factors expected to map independently onto the cucumber genome. Using the SNP array and the phenotypic data two major QTLs for fruit size of cucumber were mapped in very high accuracy (around 300 Kb) with large set of markers that should facilitate identification and cloning of major genes that contribute to fruit size in cucumber. In addition, a highly accurate haplotype map of all RILS was created to allow fine mapping of other traits segregating in this population. A detailed cucumber genetic map with 6000 markers was also established (currently the most detailed genetic map of cucumber). The integration of genetics physiology and genomic approaches in this project yielded new major infrastructure tools that can be used for understanding fruit size and many other traits of importance in cucumber. The SNP array and genetic population with an ultra-fine map can be used for future breeding efforts, high resolution mapping and cloning of traits of interest that segregate in this population. The genetic map that was developed can be used for other breeding efforts in other populations. The study of fruit development that was done during this project will be important in dissecting function of genes that that contribute to the fruit size QTLs. The SNP array can be used as tool for mapping different traits in cucumber. The development of the tools and knowledge will thus promote genetic improvement of cucumber and related cucurbits.
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3

Weller, Joel I., Harris A. Lewin, and Micha Ron. Determination of Allele Frequencies for Quantitative Trait Loci in Commercial Animal Populations. United States Department of Agriculture, February 2005. http://dx.doi.org/10.32747/2005.7586473.bard.

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Individual loci affecting economic traits in dairy cattle (ETL) have been detected via linkage to genetic markers by application of the granddaughter design in the US population and the daughter design in the Israeli population. From these analyses it is not possible to determine allelic frequencies in the population at large, or whether the same alleles are segregating in different families. We proposed to answer this question by application of the "modified granddaughter design", in which granddaughters with a common maternal grandsire are both genotyped and analyzed for the economic traits. The objectives of the proposal were: 1) to fine map three segregating ETL previously detected by a daughter design analysis of the Israeli dairy cattle population; 2) to determine the effects of ETL alleles in different families relative to the population mean; 3) for each ETL, to determine the number of alleles and allele frequencies. The ETL on Bostaurusautosome (BT A) 6 chiefly affecting protein concentration was localized to a 4 cM chromosomal segment centered on the microsatellite BM143 by the daughter design. The modified granddaughter design was applied to a single family. The frequency of the allele increasing protein percent was estimated at 0.63+0.06. The hypothesis of equal allelic frequencies was rejected at p<0.05. Segregation of this ETL in the Israeli population was confirmed. The genes IBSP, SPP1, and LAP3 located adjacent to BM143 in the whole genome cattle- human comparative map were used as anchors for the human genome sequence and bovine BAC clones. Fifteen genes within 2 cM upstream of BM143 were located in the orthologous syntenic groups on HSA4q22 and HSA4p15. Only a single gene, SLIT2, was located within 2 cM downstream of BM143 in the orthologous HSA4p15 region. The order of these genes, as derived from physical mapping of BAC end sequences, was identical to the order within the orthologous syntenic groups on HSA4: FAM13A1, HERC3. CEB1, FLJ20637, PP2C-like, ABCG2, PKD2. SPP, MEP, IBSP, LAP3, EG1. KIAA1276, HCAPG, MLR1, BM143, and SLIT2. Four hundred and twenty AI bulls with genetic evaluations were genotyped for 12 SNPs identified in 10 of these genes, and for BM143. Seven SNPs displayed highly significant linkage disequilibrium effects on protein percentage (P<0.000l) with the greatest effect for SPP1. None of SNP genotypes for two sires heterozygous for the ETL, and six sires homozygous for the ETL completely corresponded to the causative mutation. The expression of SPP 1 and ABCG2 in the mammary gland corresponded to the lactation curve, as determined by microarray and QPCR assays, but not in the liver. Anti-sense SPP1 transgenic mice displayed abnormal mammary gland differentiation and milk secretion. Thus SPP 1 is a prime candidate gene for this ETL. We confirmed that DGAT1 is the ETL segregating on BTA 14 that chiefly effects fat concentration, and that the polymorphism is due to a missense mutation in an exon. Four hundred Israeli Holstein bulls were genotyped for this polymorphism, and the change in allelic frequency over the last 20 years was monitored.
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Fluhr, Robert, and Volker Brendel. Harnessing the genetic diversity engendered by alternative gene splicing. United States Department of Agriculture, December 2005. http://dx.doi.org/10.32747/2005.7696517.bard.

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Our original objectives were to assess the unexplored dimension of alternative splicing as a source of genetic variation. In particular, we sought to initially establish an alternative splicing database for Arabidopsis, the only plant for which a near-complete genome has been assembled. Our goal was to then use the database, in part, to advance plant gene prediction programs that are currently a limiting factor in annotating genomic sequence data and thus will facilitate the exploitation of the ever increasing quantity of raw genomic data accumulating for plants. Additionally, the database was to be used to generate probes for establishing high-throughput alternative transcriptome analysis in the form of a splicing-specific oligonucleotide microarray. We achieved the first goal and established a database and web site termed Alternative Splicing In Plants (ASIP, http://www.plantgdb.org/ASIP/). We also thoroughly reviewed the extent of alternative splicing in plants (Arabidopsis and rice) and proposed mechanisms for transcript processing. We noted that the repertoire of plant alternative splicing differs from that encountered in animals. For example, intron retention turned out to be the major type. This surprising development was proven by direct RNA isolation techniques. We further analyzed EST databases available from many plants and developed a process to assess their alternative splicing rate. Our results show that the lager genome-sized plant species have enhanced rates of alternative splicing. We did advance gene prediction accuracy in plants by incorporating scoring for non-canonical introns. Our data and programs are now being used in the continuing annotation of plant genomes of agronomic importance, including corn, soybean, and tomato. Based on the gene annotation data developed in the early part of the project, it turned out that specific probes for different exons could not be scaled up to a large array because no uniform hybridization conditions could be found. Therefore, we modified our original objective to design and produce an oligonucleotide microarray for probing alternative splicing and realized that it may be reasonable to investigate the extent of alternative splicing using novel commercial whole genome arrays. This possibility was directly examined by establishing algorithms for the analysis of such arrays. The predictive value of the algorithms was then shown by isolation and verification of alternative splicing predictions from the published whole genome array databases. The BARD-funded work provides a significant advance in understanding the extent and possible roles of alternative splicing in plants as well as a foundation for advances in computational gene prediction.
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5

Jorgensen, Frieda, Andre Charlett, Craig Swift, Anais Painset, and Nicolae Corcionivoschi. A survey of the levels of Campylobacter spp. contamination and prevalence of selected antimicrobial resistance determinants in fresh whole UK-produced chilled chickens at retail sale (non-major retailers). Food Standards Agency, June 2021. http://dx.doi.org/10.46756/sci.fsa.xls618.

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Campylobacter spp. are the most common bacterial cause of foodborne illness in the UK, with chicken considered to be the most important vehicle for this organism. The UK Food Standards Agency (FSA) agreed with industry to reduce Campylobacter spp. contamination in raw chicken and issued a target to reduce the prevalence of the most contaminated chickens (those with more than 1000 cfu per g chicken neck skin) to below 10 % at the end of the slaughter process, initially by 2016. To help monitor progress, a series of UK-wide surveys were undertaken to determine the levels of Campylobacter spp. on whole UK-produced, fresh chicken at retail sale in the UK. The data obtained for the first four years was reported in FSA projects FS241044 (2014/15) and FS102121 (2015 to 2018). The FSA has indicated that the retail proxy target for the percentage of highly contaminated raw whole retail chickens should be less than 7% and while continued monitoring has demonstrated a sustained decline for chickens from major retailer stores, chicken on sale in other stores have yet to meet this target. This report presents results from testing chickens from non-major retailer stores (only) in a fifth survey year from 2018 to 2019. In line with previous practise, samples were collected from stores distributed throughout the UK (in proportion to the population size of each country). Testing was performed by two laboratories - a Public Health England (PHE) laboratory or the Agri-Food & Biosciences Institute (AFBI), Belfast. Enumeration of Campylobacter spp. was performed using the ISO 10272-2 standard enumeration method applied with a detection limit of 10 colony forming units (cfu) per gram (g) of neck skin. Antimicrobial resistance (AMR) to selected antimicrobials in accordance with those advised in the EU harmonised monitoring protocol was predicted from genome sequence data in Campylobacter jejuni and Campylobacter coli isolates The percentage (10.8%) of fresh, whole chicken at retail sale in stores of smaller chains (for example, Iceland, McColl’s, Budgens, Nisa, Costcutter, One Stop), independents and butchers (collectively referred to as non-major retailer stores in this report) in the UK that are highly contaminated (at more than 1000 cfu per g) with Campylobacter spp. has decreased since the previous survey year but is still higher than that found in samples from major retailers. 8 whole fresh raw chickens from non-major retailer stores were collected from August 2018 to July 2019 (n = 1009). Campylobacter spp. were detected in 55.8% of the chicken skin samples obtained from non-major retailer shops, and 10.8% of the samples had counts above 1000 cfu per g chicken skin. Comparison among production plant approval codes showed significant differences of the percentages of chicken samples with more than 1000 cfu per g, ranging from 0% to 28.1%. The percentage of samples with more than 1000 cfu of Campylobacter spp. per g was significantly higher in the period May, June and July than in the period November to April. The percentage of highly contaminated samples was significantly higher for samples taken from larger compared to smaller chickens. There was no statistical difference in the percentage of highly contaminated samples between those obtained from chicken reared with access to range (for example, free-range and organic birds) and those reared under standard regime (for example, no access to range) but the small sample size for organic and to a lesser extent free-range chickens, may have limited the ability to detect important differences should they exist. Campylobacter species was determined for isolates from 93.4% of the positive samples. C. jejuni was isolated from the majority (72.6%) of samples while C. coli was identified in 22.1% of samples. A combination of both species was found in 5.3% of samples. C. coli was more frequently isolated from samples obtained from chicken reared with access to range in comparison to those reared as standard birds. C. jejuni was less prevalent during the summer months of June, July and August compared to the remaining months of the year. Resistance to ciprofloxacin (fluoroquinolone), erythromycin (macrolide), tetracycline, (tetracyclines), gentamicin and streptomycin (aminoglycosides) was predicted from WGS data by the detection of known antimicrobial resistance determinants. Resistance to ciprofloxacin was detected in 185 (51.7%) isolates of C. jejuni and 49 (42.1%) isolates of C. coli; while 220 (61.1%) isolates of C. jejuni and 73 (62.9%) isolates of C. coli isolates were resistant to tetracycline. Three C. coli (2.6%) but none of the C. jejuni isolates harboured 23S mutations predicting reduced susceptibility to erythromycin. Multidrug resistance (MDR), defined as harbouring genetic determinants for resistance to at least three unrelated antimicrobial classes, was found in 10 (8.6%) C. coli isolates but not in any C. jejuni isolates. Co-resistance to ciprofloxacin and erythromycin was predicted in 1.7% of C. coli isolates. 9 Overall, the percentages of isolates with genetic AMR determinants found in this study were similar to those reported in the previous survey year (August 2016 to July 2017) where testing was based on phenotypic break-point testing. Multi-drug resistance was similar to that found in the previous survey years. It is recommended that trends in AMR in Campylobacter spp. isolates from retail chickens continue to be monitored to realise any increasing resistance of concern, particulary to erythromycin (macrolide). Considering that the percentage of fresh, whole chicken from non-major retailer stores in the UK that are highly contaminated (at more than 1000 cfu per g) with Campylobacter spp. continues to be above that in samples from major retailers more action including consideration of interventions such as improved biosecurity and slaughterhouse measures is needed to achieve better control of Campylobacter spp. for this section of the industry. The FSA has indicated that the retail proxy target for the percentage of highly contaminated retail chickens should be less than 7% and while continued monitoring has demonstrated a sustained decline for chickens from major retailer stores, chicken on sale in other stores have yet to meet this target.
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6

Vakharia, Vikram, Shoshana Arad, Yonathan Zohar, Yacob Weinstein, Shamila Yusuff, and Arun Ammayappan. Development of Fish Edible Vaccines on the Yeast and Redmicroalgae Platforms. United States Department of Agriculture, February 2013. http://dx.doi.org/10.32747/2013.7699839.bard.

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Betanodaviruses are causative agents of viral nervous necrosis (VNN), a devastating disease of cultured marine fish worldwide. Betanodavirus (BTN) genome is composed of two single-stranded, positive-sense RNA molecules. The larger genomic segment, RNA1 (3.1 kb), encodes the RNA-dependent RNA polymerase, while the smaller genomic segment, RNA 2 (1.4kb), encodes the coat protein. This structural protein is the host-protective antigen of VNN which assembles to form virus-like particles (VLPs). BTNs are classified into four genotypes, designated red-spotted grouper nervous necrosis virus (RGNNV), barfin flounder nervous necrosis virus (BFNNV), tiger puffer nervous necrosis virus (TPNNV), and striped jack nervous necrosis virus (SJNNV), based on phylogenetic analysis of the coat protein sequences. RGNNV type is quite important as it has a broad host-range, infecting warm-water fish species. At present, there is no commercial vaccine available to prevent VNN in fish. The general goal of this research was to develop oral fish vaccines in yeast and red microalgae (Porphyridium sp.) against the RGNNV genotype. To achieve this, we planned to clone and sequence the coat protein gene of RGNNV, express the coat protein gene of RGNNV in yeast and red microalgae and evaluate the immune response in fish fed with recombinantVLPs antigens produced in yeast and algae. The collaboration between the Israeli group and the US group, having wide experience in red microalgae biochemistry, molecular genetics and large-scale cultivation, and the development of viral vaccines and eukaryotic protein expression systems, respectively, was synergistic to produce a vaccine for fish that would be cost-effective and efficacious against the betanodavirus infection.
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Lee, Richard, Moshe Bar-Joseph, K. S. Derrick, Aliza Vardi, Roland Brlansky, Yuval Eshdat, and Charles Powell. Production of Antibodies to Citrus Tristeza Virus in Transgenic Citrus. United States Department of Agriculture, September 1995. http://dx.doi.org/10.32747/1995.7613018.bard.

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Citrus tristeza virus (CTV) is the most important virus disease of citrus in the world. CTV causes death of trees on sour orange rootstock and/or stem pitting of scions regardless of rootstock which results in trees of low vigor, reduced yield with reduction in size and quality of fruit. The purpose of this project was to produce monoclonal antibodies (MABs) to CTV coat protein (CP), develop single domain antibodies (dAbs) or Fab fragments which neutralize the infection by binding to the virus, and to produce transformed plants which express the dAbs. The objectives of this research have been met and putative transgenic tobacco and citrus plants have been developed. These putative transgenic plants are presently undergoing evaluation to determine the level of dAbs expression and to determine their resistance to CTV. Additionally, the CTV genome has been sequenced and the CP gene of several biologically characterized CTV strains molecular characterized. This has indicated a correlation between CP sequence homology and biological activity, and the finding of DI RNAs associated with some CTV strains. Several MABs have been produced which enable broad spectrum identification of CTV strains while other MABs enable differentiation between mild and severe strains. The use of selected MAbs and determination of the CP gene sequence has enabled predictions of biological activities of unknown CTV isolates. The epitopes of two MABs, one reacting selectively with severe CTV strains and the other reacting with all strains, have been characterized at the molecular level.
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8

Newton, Ronald, Joseph Riov, and John Cairney. Isolation and Functional Analysis of Drought-Induced Genes in Pinus. United States Department of Agriculture, September 1993. http://dx.doi.org/10.32747/1993.7568752.bard.

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Drought is a common factor limiting timber production in the U.S. and Israel. Loblolly (Pinus taeda) and alleppo pine (Pinus halepensis) seedling survival is reduced when out planted, and growth and reproduction are often hindered by periodic droughts during later stages of tree development. Molecular and gene responses to drought stress have not been characterized. The objectives were to characterize drought-induced gene clones from these pines, to determine the effects of a growth regulator on drought tolerance, ABA levels, and drought-induced gene expression in alleppo pine, and to develop procedures for loblolly pine transformation. Nearly 20 cDNA clones influenced by gradual, prolonged drought stress have been isolated. Many of these have been shown to be induced by drought stress, whereas several others are down-regulated. These are the first drought-induced genes isolated from a pine species. Two genomic clones (lp5-1 and lp3-1) have been sequenced and characterized, and each has been found to be associated with a gene family. Clone lp5 appears to code for a cell wall protein, and clone lp3 codes for a nuclear protein. The former may be associated with changing the elastic properties of the cell wall, while the latter may be involved in signal transduction and/or protection from desiccation in the nucleus. Clone lp3 is similar to a drought-induced gene from tomato and is regulated by ABA. Several DNA sequences that are specific to induction during growth-retardation in alleppo pine by uniconazole have been identified. The active DNA species is now being identified. Promoters from genomic clones, lp3 and lp5, have been sequenced. Both are functional when fused with the gus reporter gene and transferred to other plant tissues as well as responding to a simulated drought stress. Through exodeletion analysis, it has been established that the promoter ABRE element of lp3 responds to ABA and that drought-induction of lp3 expression may also involve ABA. Stable tobacco transformants carrying either the lp5 or the lp3 promoter fused to a reporter gus gene have been obtained. The lp5lgus fusion was expressed at several stages of tobacco development and differentiation including the reproductive stage. There was no difference in phenotype between the transformants and the wild type. Embryogenesis procedures were developed for slash pine, but attempts to couple this process with gene transfer and plantlet transformation were not successful. Transformation of pine using Agrobacterium appears tractable, but molecular data supporting stable integration of the Agrobacterium-transferred gene are still inconclusive.
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Jordan, Ramon L., Abed Gera, Hei-Ti Hsu, Andre Franck, and Gad Loebenstein. Detection and Diagnosis of Virus Diseases of Pelargonium. United States Department of Agriculture, July 1994. http://dx.doi.org/10.32747/1994.7568793.bard.

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Pelargonium (Geranium) is the number one pot plant in many areas of the United States and Europe. Israel and the U.S. send to Europe rooted cuttings, foundation stocks and finished plants to supply a certain share of the market. Geraniums are propagated mainly vegetatively from cuttings. Consequently, viral diseases have been and remain a major threat to the production and quality of the crop. Among the viruses isolated from naturally infected geraniums, 11 are not specific to Pelargonium and occur in other crops while 6 other viruses seem to be limited to geranium. However, several of these viruses are not sufficiently characterized to conclude that they are distinct agents and their nomenclature and taxonomy are confusing. The ability to separate, distinguish and detect the different viruses in geranium will overcome obstacles te developing effective detection and certification schemes. Our focus was to further characterize some of these viruses and develop better methods for their detection and control. These viruses include: isolates of pelargonium line pattern virus (PLPV), pelargonium ringspot virus (PelRSV), pelargonium flower break virus (PFBV), pelargonium leaf curl (PLCV), and tomato ringspot virus (TomRSV). Twelve hybridoma cell lines secreting monoclonal antibodies specific to a geranium isolate of TomRSV were produced. These antibodies are currently being characterized and will be tested for the ability to detect TomRSV in infected geraniums. The biological, biochemical and serological properties of four isometric viruses - PLPV, PelRSV, and PFBV (and a PelRSV-like isolate from Italy called GR57) isolated from geraniums exhibiting line and ring pattern or flower break symptoms - and an isolate ol elderbeny latent virus (ELV; which the literature indicates is the same as PelRSV) have been determined Cloned cDNA copies of the genomic RNAs of these viruses were sequenced and the sizes and locations of predicted viral proteins deduced. A portion of the putative replicase genes was also sequenced from cloned RT-PCR fragments. We have shown that, when compared to the published biochemical and serological properties, and sequences and genome organizations of other small isometric plant viruses, all of these viruses should each be considered new, distinct members of the Carmovirus group of the family Tombusviridae. Hybridization assays using recombinant DNA probes also demonstrated that PLPV, PelRSV, and ELV produce only one subgenomic RNA in infected plants. This unusual property of the gene expression of these three viruses suggests that they are unique among the Carmoviruses. The development of new technologies for the detection of these viruses in geranium was also demonstrated. Hybridization probes developed to PFBV (radioactively-labeled cRNA riboprobes) and to PLPV (non-radioactive digoxigenin-labeled cDNAs) were generally shown to be no more sensitive for the detection of virus in infected plants than the standard ELISA serology-based assays. However, a reverse transcriptase-polymerase chain reaction assay was shown to be over 1000 times more sensitive in detecting PFBV in leaf extracts of infected geranium than was ELISA. This research has lead to a better understanding of the identity of the viruses infecting pelargonium and to the development of new tools that can be used in an improved scheme of providing virus-indexed pelargonium plants. The sequence information, and the serological and cloned DNA probes generated from this work, will allow the application of these new tools for virus detection, which will be useful in domestic and international indexing programs which are essential for the production of virus-free germplasm both for domestic markets and the international exchange of plant material.
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Rodriguez, Russell, and Stanley Freeman. Characterization of fungal symbiotic lifestyle expression in Colletotrichum and generating non-pathogenic mutants that confer disease resistance, drought tolerance, and growth enhancement to plant hosts. United States Department of Agriculture, February 2005. http://dx.doi.org/10.32747/2005.7587215.bard.

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Fungal plant pathogens are responsible for extensive annual crop and revenue losses throughout the world. To better understand why fungi cause diseases, we performed gene-disruption mutagenesis on several pathogenic Colletotrichum species and demonstrated that pathogenic isolates can be converted to symbionts (mutualism, commensalism, parasitism) expressing non-pathogenic lifestyles. The objectives of this proposal were to: 1- generate crop-specific mutants by gene disruption that express mutualistic lifestyles, 2- assess the ability of the mutualists to confer disease resistance, drought tolerance, and growth enhancement to host plants, 3- compare fslm1 sequences and their genomic locations in the different species, and 4- document the colonization process of each Colletotrichum species.It was demonstrated that wildtype pathogenic Colletotrichum isolates, can be converted by mutation from expressing a pathogenic lifestyle to symbionts expressing non-pathogenic lifestyles. In the US, mutants of Colletotrichum were isolated by homologous gene disruption using a vector containing a disrupted FSlm1 sequence while in Israel, C. acutatum mutants were selected by restriction enzyme mediated integration (REMI) transformation. One group (US) of non-pathogenic mutants conferred disease protection against pathogenic species of Colletotrichum, Fusarium, and Phytophthora; drought tolerance; and growth enhancement to host plants. These mutants were defined as mutualists and disease resistance correlated to a decrease in the time required for hosts to activate defense systems when exposed to virulent fungi. The second group (Israel) of non-pathogenic mutants did not confer disease resistance and were classified as commensals. In addition, we demonstrated that wildtype pathogenic Colletotrichum species can express non-pathogenic lifestyles, including mutualism, on plants they colonize asymptomatically. The expected long term contribution of this research to agriculture in the US and Israel is threefold. Host-specific mutualists will be utilized in the various crops to confer (1) disease resistance to reduce dependence on chemical fungicides; (2) drought tolerance to reduce water consumption for irrigation; (3) growth enhancement to increase yields.
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