Search results for: phylogenetic relationship
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6732

Search results for: phylogenetic relationship

6702 Prevalence and Pathomorphological Study of Natural Coccidiosis in Japanese Quails (Coturnix coturnix japonica) in Iran

Authors: M. Khordadmehr, V. R. Ranjbar, R. Norouzi, M. Zeinoddin

Abstract:

Coccidiosis is recognized as a serious parasitic disease problem limiting quail industry recently. But the data on incidence, clinical signs, species of coccidia and pathological changes in Japanese quail are rare, especially in Iran in spite of the significant improvement of commercial quail breeding in this country in recent decades. Therefore, in the present paper was studied natural infection of quail coccidiosis in three commercial rearing farms with 80% morbidity and 3% mortality rate. For this purpose, fecal sample, oocyst examination, and morphological study were performed beside necropsy, histopathology, and PCR to confirm the diagnosis. In the affected birds, clinical signs included brown diarrhea, weakness, and pale face. In the fecal examination, three species of the genus Eimeria were identified including E. uzura, E. bateri, and E. tsunodai. At necropsy, the main gross lesions were edema, congestion and small blood spots in the small intestine. In histopathologic examination, endogenous stages of the parasites associated with hyperplasia of the intestinal glands, mild congestion, infiltration of mononuclear cells, and edema were observed in the intestine. The molecular study using BSEF and BSER specific primers confirmed the presence of the genus Eimeria in the affected birds. Interestingly, phylogenetic analysis showed relatively high bootstrap values in Japanese quail Eimeria with E. acervuline and E. maxima strains in the chicken. The present study is the first phylogenetic findings on Eimeria of quail which could be valuable for further research on Japanese quail coccidiosis.

Keywords: coccidiosis, Japanese Quail, pathomorphology, phylogenetic analysis

Procedia PDF Downloads 244
6701 Molecular Epidemiology of Circulating Adenovirus Types in Acute Conjunctivitis Cases in Chandigarh, North India

Authors: Mini P. Singh, Jagat Ram, Archit Kumar, Tripti Rungta, Jasmine Khurana, Amit Gupta, R. K. Ratho

Abstract:

Introduction: Human adenovirus is the most common agent involved in viral conjunctivitis. The clinical manifestations vary with different serotypes. The identification of the circulating strains followed by phylogenetic analysis can be helpful in understanding the origin and transmission of the disease. The present study aimed to carry out molecular epidemiology of the adenovirus types in the patients with conjunctivitis presenting to the eye centre of a tertiary care hospital in North India. Materials and Methods: The conjunctival swabs were collected from 23 suspected adenoviral conjunctivitis patients between April-August, 2014 and transported in viral transport media. The samples were subjected to nested PCR targeting hexon gene of human adenovirus. The band size of 956bp was eluted and 8 representative positive samples were subjected to sequencing. The sequences were analyzed by using CLUSTALX2.1 and MEGA 5.1 software. Results: The male: female ratio was found to be 3.6:1. The mean age of presenting patients was 43.95 years (+17.2). Approximately 52.1% (12/23) of patients presented with bilateral involvement while 47.8% (11/23) with unilateral involvement of the eye. Human adenovirus DNA could be detected in 65.2% (15/23) of the patients. The phylogenetic analysis revealed presence of serotype 8 in 7 patients and serotype 4 in one patient. The serotype 8 sequences showed 99-100% identity with Tunisian, Indian and Japanese strains. The adenovirus serotype 4 strains had 100% identity with strains from Tunisia, China and USA. Conclusion: Human adenovirus was found be an important etiological agent for conjunctivitis in our set up. The phylogenetic analysis showed that the predominant circulating strains in our epidemic keratoconjunctivitis were serotypes 8 and 4.

Keywords: conjunctivitis, human adenovirus, molecular epidemiology, phylogenetics

Procedia PDF Downloads 279
6700 Elongation Factor 1 Alpha Molecular Phylogenetic Analysis for Anastrepha fraterculus Complex

Authors: Pratibha Srivastava, Ayyamperumal Jeyaprakash, Gary Steck

Abstract:

Exotic, invasive tephritid fruit flies (Diptera: Tephritidae) are a major concern to fruit and vegetable production in the USA. Timely detection and identification of these agricultural pests facilitate the possibility of eradication from newly invaded areas. They spread primarily as larvae in infested fruits carried in commerce or personal baggage. Identification of larval stages to species level is difficult but necessary to determine pest loads and their pathways into the USA. The main focus of this study is the New World genus, Anastrepha. Many of its constituent taxa are pests of major economic importance. This study is significant for national quarantine use, as morphological diagnostics to separate larvae of the various members remain poorly developed. Elongation factor 1 alpha sequences were amplified from Anastrepha fraterculus specimens collected from South America (Ecuador and Peru). Phylogenetic analysis was performed to characterize the Anastrepha fraterculus complex at a molecular level.

Keywords: anastrepha, diptera, elongation factor, fruit fly

Procedia PDF Downloads 205
6699 Phylogenetic Analysis of Georgian Populations of Potato Cyst Nematodes Globodera Rostochiensis

Authors: Dali Gaganidze, Ekaterine Abashidze

Abstract:

Potato is one of the main agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. In traditional potato growing regions (Svaneti, Samckhet javaheti and Tsalka), the yield is higher than 30-35 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (PCN) are harmful around the world. Yield losses caused by PCN are estimated up to 30%. Rout surveys conducted in two geographically distinct regions of Georgia producing potatoes - Samtskhe - Javakheti and Svaneti revealed potato cyst nematode Globodera rostochiensi. The aim of the study was the Phylogenetic analyses of Globodera rostochiensi revealed in Georgia by the amplification and sequencing of 28S gen in the D3 region and intergenic ITS1-15.8S-ITS2 region. Identification of all the samples from the two Globodera populations (Samtskhe - Javakheti and Svaneti), i.e., G. rostochiensis (20 isolates) were confirmed by conventional multiplex PCR with ITS 5 universal and PITSp4, PITSr3 specific primers of the cyst nematodes’ (G. pallida, G. rostochiensis). The size of PCR fragment 434 bp confirms that PCN samples from two populations, Samtskhe- Javakheti and Svaneti, belong to G. rostochiensi . The ITS1–5.8S-ITS2 regions were amplified using prime pairs: rDNA1 ( 5’ -TTGATTACGTCCCTGCCCTTT-3’ and rDNA2( 5’ TTTCACTCGCCGTTACTAAGG-3’), D3 expansion regions were amplified using primer pairs: D3A (5’ GACCCCTCTTGAAACACGGA-3’) and D3B (5’-TCGGAAGGAACCAGCTACTA-3’. PCR products of each region were cleaned up and sequenced using an ABI 3500xL Genetic Analyzer. Obtained sequencing results were analyzed by computer program BLASTN (https://blast.ncbi.nlm.nih.gov/Blast.cg). Phylogenetic analyses to resolve the relationships between the isolates were conducted in MEGA7 using both distance- and character-based methods. Based on analysis of G.rostochiensis isolate`s D3 expansion regions are grouped in three major clades (A, B and C) on the phylogenetic tree. Clade A is divided into three subclades; clade C is divided into two subclades. Isolates from the Samtckhet-javakheti population are in subclade 1 of clade A and isolates in subclade 1 of clade C. Isolates) from Svaneti populations are in subclade 2 of clade A and in clad B. In Clade C, subclade two is presented by three isolates from Svaneti and by one isolate (GL17) from Samckhet-Javakheti. . Based on analysis of G.rostochiensis isolate`s ITS1–5.8S-ITS2 regions are grouped in two main clades, the first contained 20 Georgian isolates of Globodera rostochiensis from Svaneti . The second clade contained 15 isolates of Globodera rostochiensis from Samckhet javakheti. Our investigation showed of high genetic variation of D3 and ITS1–5.8S-ITS2 region of rDNA of the isolates of G. rostochiensis from different geographic origins (Svameti, Samckhet-Javakheti) of Georgia. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia : Project # FR17_235

Keywords: globodera rostochiensi, PCR, phylogenetic tree, sequencing

Procedia PDF Downloads 195
6698 Phylogenetic Diversity and Antibiotic Resistance in Sediments of Aegean Sea

Authors: Ilknur Tuncer, Nihayet Bizsel

Abstract:

The studies in bacterial diversity and antimicrobial resistance in coastal areas are important to understand the variability in the community structures and metabolic activities. In the present study, antimicrobial susceptibility and phylogenetic analysis of bacteria isolated from stations with different depths and influenced by terrestrial and marine fluxes in eastern Aegean Sea were illustrated. 51% of the isolates were found as resistant and 14% showed high MAR index indicating the high-risk sources of contamination in the environment. The resistance and the intermediate levels and high MAR index of the study area were 38–60%, 11–38% and 0–40%, respectively. According to 16S rRNA gene analysis, it was found that the isolates belonged to two phyla Firmicutes and Gammaproteobacteria with the genera Bacillus, Halomonas, Oceanobacillus, Photobacterium, Pseudoalteromonas, Psychrobacter, and Vibrio. 47% of Bacillus strains which were dominant among all isolates were resistant. In addition to phylogenetically diverse bacteria, the variability in resistance, intermediate and high MAR index levels of the study area indicated the effect of geographical differences.

Keywords: bacterial diversity, multiple antibiotic resistance, 16S rRNA genes, Aegean Sea

Procedia PDF Downloads 412
6697 Antibacterial Activity of Endophytic Bacteria against Multidrug-Resistant Bacteria: Isolation, Characterization, and Antibacterial Activity

Authors: Maryam Beiranvand, Sajad Yaghoubi

Abstract:

Background: Some microbes can colonize plants’ inner tissues without causing obvious damage and can even produce useful bioactive substances. In the present study, the diversity of the endophytic bacteria associated with medicinal plants from Iran was investigated by culturing techniques, molecular gene identification, as well as measuring them for antibacterial activity. Results: In the spring season from 2013 to 2014, 35 herb pharmacology samples were collected, sterilized, meshed, and then cultured on selective media culture. A total of 199 endophytic bacteria were successfully isolated from 35 tissue cultures of medical plants, and sixty-seven out of 199 bacterial isolates were subjected to identification by the 16S rRNA gene sequence analysis method. Based on the sequence similarity gene and phylogenetic analyses, these isolates were grouped into five classes, fourteen orders, seventeen families, twenty-one genera, and forty strains. The most abundant group of endophytic bacteria was actinobacterial, consisting of thirty-two (47%) out of 67 bacterial isolates. Ten (22.3%) out of 67 bacterial isolates remained unidentified and classified at the genus level. The signature of the 16S rRNA gene formed a distinct line in a phylogenetic tree showing that they might be new species of bacteria. One (5.2%) out of 67 bacterial isolates was still not well categorized. Forty-two out of 67 strains were candidates for antimicrobial activity tests. Nineteen (45%) out of 42 strains showed antimicrobial activity multidrug resistance (MDR); thirteen (68%) out of 19 strains were allocated to classes actinobacteria. Four (21%) out of 19 strains belonged to the Bacillaceae family, one (5.2%) out of 19 strains was the Paenibacillaceae family, and one (5.2%) out of 19 strains belonged to the Pseudomonadaceae family. The other twenty-three strains did not show inhibitory activities. Conclusions: Our research showed a high-level phylogenetic diversity and the intoxicating antibiotic activity of endophytic bacteria in the herb pharmacology of Iran.

Keywords: Antibacterial activity, endophytic bacteria, multidrug-resistant bacteria, whole genom sequencing

Procedia PDF Downloads 86
6696 Isolation, Characterization, and Antibacterial Activity of Endophytic Bacteria from Iranian Medicinal Plants

Authors: Maryam Beiranvand, Sajad Yaghoubi

Abstract:

Background: Some microbes can colonize plants’ inner tissues without causing obvious damage and can even produce useful bioactive substances. In the present study, the diversity of the endophytic bacteria associated with medicinal plants from Iran was investigated by culturing techniques, molecular gene identification, as well as measuring them for antibacterial activity. Results: In the spring season from 2013 to 2014, 35 herb pharmacology samples were collected, sterilized, meshed, and then cultured on selective media culture. A total of 199 endophytic bacteria were successfully isolated from 35 tissue cultures of medical plants, and sixty-seven out of 199 bacterial isolates were subjected to identification by the 16S rRNA gene sequence analysis method. Based on the sequence similarity gene and phylogenetic analyses, these isolates were grouped into five classes, fourteen orders, seventeen families, twenty-one genera, and forty strains. The most abundant group of endophytic bacteria was actinobacterial, consisting of thirty-two (47%) out of 67 bacterial isolates. Ten (22.3%) out of 67 bacterial isolates remained unidentified and classified at the genus level. The signature of the 16S rRNA gene formed a distinct line in a phylogenetic tree showing that they might be new species of bacteria. One (5.2%) out of 67 bacterial isolates was still not well categorized. Forty-two out of 67 strains were candidates for antimicrobial activity tests. Nineteen (45%) out of 42 strains showed antimicrobial activity multidrug-resistance (MDR); thirteen (68%) out of 19 strains were allocated to classes actinobacteria. Four (21%) out of 19 strains belonged to the Bacillaceae family, one (5.2%) out of 19 strains was the Paenibacillaceae family, and one (5.2%) out of 19 strains belonged to the Pseudomonadaceae family. The other twenty-three strains did not show inhibitory activities. Conclusions: Our research showed a high-level phylogenetic diversity and the intoxicating antibiotic activity of endophytic bacteria in the herb pharmacology of Iran.

Keywords: medical plant, endophytic bacteria, antimicrobial activity, whole genome sequencing analysis

Procedia PDF Downloads 124
6695 Molecular Characterization of Grain Storage Proteins in Some Hordeum Species

Authors: Manar Makhoul, Buthainah Alsalamah, Salam Lawand, Hassan Azzam

Abstract:

The major storage proteins in endosperm of 33 cultivated and wild barley genotypes (H.vulgare, H. spontaneum, H. bulbosum, H. murinum, H. marinum) were analyzed to demonstrate the variation in the hordein polypeptides encoded by multigene families in grains. The SDS-PAGE revealed 13 and 17 alleles at the Hor1 and the Hor2 loci respectively, with frequencies from 0.83 to 14 and 0.56 to 13.41% respectively, while seven alleles at the Hor3 locus with frequencies from 3.63 to 30.91% were recognized. The phylogenetic analysis indicated to relevance of the polymorphism in hordein patterns as successful tool in identifying the individual genotypes and discriminating the species according to genome type. We also reported in this research complete nucleotide sequence B-hordein genes of seven wild and cultivated barley genotypes. A 152bp upstream sequence of B-hordein promoter contained a TATA box, CATC box, AAAG motif, N-motif and E-motif. In silico analysis of B-Hordein sequences demonstrated that the coding regions were not interrupted by any intron, and included the complete ORF which varied between 882 and 906 bp, and encoded mature proteins with 293-301 residues characterized by high contents of glutamine (29%), and proline (18%). Comparison of the predicted polypeptide sequences with the published ones suggested that all S-rich prolamins genes are descended from common ancestor. The sequence started at N-terminal with a signal peptide, and then followed directly by two domains; a repetitive one based on the repetition of the repeat unit PQQPFPQQ and C-terminal domain. Also, it was found that positions of the eight cysteine residues were highly conserved in all the B-hordein sequences, but Hordeum bulbosum had additional unpaired one. The phylogenetic tree of B-hordein polypeptide separated the genotypes in distinct seven subgroups. In general, the high homology between B-hordeins and LMW glutenin subunits suggests similar bread-making influences for these B-hordeins.

Keywords: hordeum, phylogenetic tree, sequencing, storage protein

Procedia PDF Downloads 267
6694 In silico Comparative Analysis of Chloroplast Genome (cpDNA) and Some Individual Genes (rbcL and trnH-psbA) in Pooideae Subfamily Members

Authors: Ibrahim Ilker Ozyigit, Ertugrul Filiz, Ilhan Dogan

Abstract:

An in silico analysis of Brachypodium distachyon, Triticum aestivum, Festuca arundinacea, Lolium perenne, Hordeum vulgare subsp. vulgare of the Pooideaea was performed based on complete chloroplast genomes including rbcL coding and trnH-psbA intergenic spacer regions alone to compare phylogenetic resolving power. Neighbor-joining, Minimum Evolution, and Unweighted Pair Group Method with arithmetic mean methods were used to reconstruct phylogenies with the highest bootstrap supported the obtained data from whole chloroplast genome sequence. The highest and lowest values from nucleotide diversity (π) analysis were found to be 0.315813 and 0.043495 in rbcL coding region in chloroplast genome and complete chloroplast genome, respectively. The highest transition/transversion bias (R) value was recorded as 1.384 in complete chloroplast genomes. F. arudinacea-L. perenne clade was uncovered in all phylogenies. Sequences of rbcL and trnH-psbA regions were not able to resolve the Pooideae phylogenies due to lack of genetic variation.

Keywords: chloroplast DNA, Pooideae, phylogenetic analysis, rbcL, trnH-psbA

Procedia PDF Downloads 379
6693 Bioinformatic Study of Follicle Stimulating Hormone Receptor (FSHR) Gene in Different Buffalo Breeds

Authors: Hamid Mustafa, Adeela Ajmal, Kim EuiSoo, Noor-ul-Ain

Abstract:

World wild, buffalo production is considered as most important component of food industry. Efficient buffalo production is related with reproductive performance of this species. Lack of knowledge of reproductive efficiency and its related genes in buffalo species is a major constraint for sustainable buffalo production. In this study, we performed some bioinformatics analysis on Follicle Stimulating Hormone Receptor (FSHR) gene and explored the possible relationship of this gene among different buffalo breeds and with other farm animals. We also found the evolution pattern for this gene among these species. We investigate CDS lengths, Stop codon variation, homology search, signal peptide, isoelectic point, tertiary structure, motifs and phylogenetic tree. The results of this study indicate 4 different motif in this gene, which are Activin-recp, GS motif, STYKc Protein kinase and transmembrane. The results also indicate that this gene has very close relationship with cattle, bison, sheep and goat. Multiple alignment (MA) showed high conservation of motif which indicates constancy of this gene during evolution. The results of this study can be used and applied for better understanding of this gene for better characterization of Follicle Stimulating Hormone Receptor (FSHR) gene structure in different farm animals, which would be helpful for efficient breeding plans for animal’s production.

Keywords: buffalo, FSHR gene, bioinformatics, production

Procedia PDF Downloads 532
6692 Smelling Our Way through Names: Understanding the Potential of Floral Volatiles as Taxonomic Traits in the Fragrant Ginger Genus Hedychium

Authors: Anupama Sekhar, Preeti Saryan, Vinita Gowda

Abstract:

Plants, due to their sedentary lifestyle, have evolved mechanisms to synthesize a huge diversity of complex, specialized chemical metabolites, a majority of them being volatile organic compounds (VOCs). These VOCs are heavily involved in their biotic and abiotic interactions. Since chemical composition could be under the same selection processes as other morphological characters, we test if VOCs can be used to taxonomically distinguish species in the well-studied, fragrant ginger genus -Hedychium (Zingiberaceae). We propose that variations in the volatile profiles are suggestive of adaptation to divergent environments, and their presence could be explained by either phylogenetic conservatism or ecological factors. In this study, we investigate the volatile chemistry within Hedychium, which is endemic to Asian palaeotropics. We used an unsupervised clustering approach which clearly distinguished most taxa, and we used ancestral state reconstruction to estimate phylogenetic signals and chemical trait evolution in the genus. We propose that taxonomically, the chemical composition could aid in species identification, especially in species complexes where taxa are not morphologically distinguishable, and extensive, targeted chemical libraries will help in this effort.

Keywords: chemotaxonomy, dynamic headspace sampling, floral fragrance, floral volatile evolution, gingers, Hedychium

Procedia PDF Downloads 95
6691 Multilocus Phylogenetic Approach Reveals Informative DNA Barcodes for Studying Evolution and Taxonomy of Fusarium Fungi

Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

Abstract:

Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

Procedia PDF Downloads 324
6690 Positive Interactions among Plants in Pinegroves over Quarzitic Sands

Authors: Enrique González Pendás, Vidal Pérez Hernández, Jorge Ferro Díaz, Nelson Careaga Pendás

Abstract:

The investigation is carried out on the Protected Area of San Ubaldo, toward the interior of an open pinegrove with palm trees in a dry plainness of quar zitic sands, belonging to the Floristic Managed Reservation San Ubaldo-Sabanalamar, Guane, Pinar del Río, Cuba. This area is characterized by drastic seasonal variations, high temperatures and water evaporation, strong solar radiation, with sandy soils of almost pure quartz, which are very acid and poor in nutrients. The objective of the present work is to determine evidence of facilitation and its relationship with the structure and composition of plant communities in these peculiar ecosystems. For this study six lineal parallel transepts of 100 m are traced, in those, a general recording of the flora is carried out. To establish which plants act as nurses, is taken into account a height over 1 meter, canopy over 1.5 meter and the occurrence of several species under it. Covering was recorded using the line intercept method; the medium values of species richness for the taxa under nurses is compared with those that are located in open spaces among them. Then, it is determined which plants are better recruiter of other species (better nurses). An experiment is made to measure and compare some parameters in pine seedlings under the canopy of the Byrsonima crassifolia (L.) Kunth. and in open spaces, also the number of individuals is counted by species to calculate the frequency and total abundance in the study area. As a result, it is offered an up-to-date floristic list, a phylogenetic tree of the plant community showing a high phylodiversity, it is proven that the medium values of species richness and abundance of species under the nurses, is significantly superior to those occurring in open spaces. Furthermore, by means of phylogenetic trees it is shown that the species which cohabit under the nurses are not phylogenetically related. The former results are cited evidences of facilitation among plants, as well as it is one more time shown the importance of the nurse effect in preserving plant diversity on extreme environments.

Keywords: facilitation, nurse plants, positive interactions, quarzitic sands

Procedia PDF Downloads 341
6689 Cysteine Proteases of Plants That Act on the Coagulation Cascade: Approach from Bioinformatics

Authors: Tapiwa Brine Mutsauri

Abstract:

The MEROPS system is an information resource for proteases that classifies them into clans according to their catalytic type. Within the Plant kingdom, cysteine ​​proteases are one of the best known, as they are the catalytic type on which the first studies on plant proteases were focused. Plant cysteine ​​proteases have a similar mechanism of action to serine proteases, and some are known to have activity on factors of the blood coagulation cascade, such as a potent antithrombotic effect, and also cause increased fibrinolysis. Of a few plant cysteine ​​proteases, the three-dimensional structure is known, so a method of interest to be able to predict their potential activity on the factors of the coagulation cascade would be to know their structure. Phylogenetics is the study of the evolutionary relationships between biological entities, often species, individuals, or genes (which can be called taxa). It is essential to identify the evolutionary position and the possible distribution of these enzymes in the plant kingdom, particularly those that act on coagulation factors. Bioinformatic tools, such as Clustal Omega / Jalview and Mega6, can be used to create phylogenetic trees. From the results of the alignment, it can be seen that although there is a certain degree of conservation (Conservation) and consensus (Consensus) among the eleven sequences, the functionally important motifs (those corresponding to the active site), the degree of conservation and consensus is very low. We could then infer that although activity on coagulation is reported for these enzymes, linked to their structural and mechanistic similarity with serine proteases, this activity may not have a direct or primary relationship with the proteolytic activity associated with their common, poorly conserved active site in this case. This ultimately could be related to modifications in the reaction mechanism of several of the enzymes studied, which would require more detailed study. Also, below, we will deal with factors that may have a greater influence on this result. The results of this work enrich the understanding of how species (and molecular sequences in general) evolve. Through phylogenetics, we learn not only how sequences came to be the way they are today but also the general principles that allow us to predict how they will change in the future. For pharmaceutical sciences, phylogenetic selection of biologically related species can help identify closely related members of a species with compounds of pharmacological importance, such as plant cysteine ​​proteases, in addition to identifying structural features that may influence their pharmacological activity and which can be valuable for drug design.

Keywords: computational simulation, proteases, coagulation, bioinformatics

Procedia PDF Downloads 17
6688 Broad Survey of Fine Root Traits to Investigate the Root Economic Spectrum Hypothesis and Plant-Fire Dynamics Worldwide

Authors: Jacob Lewis Watts, Adam F. A. Pellegrini

Abstract:

Prairies, grasslands, and forests cover an expansive portion of the world’s surface and contribute significantly to Earth’s carbon cycle. The largest driver of carbon dynamics in some of these ecosystems is fire. As the global climate changes, most fire-dominated ecosystems will experience increased fire frequency and intensity, leading to increased carbon flux into the atmosphere and soil nutrient depletion. The plant communities associated with different fire regimes are important for reassimilation of carbon lost during fire and soil recovery. More frequent fires promote conservative plant functional traits aboveground; however, belowground fine root traits are poorly explored and arguably more important drivers of ecosystem function as the primary interface between the soil and plant. The root economic spectrum (RES) hypothesis describes single-dimensional covariation between important fine-root traits along a range of plant strategies from acquisitive to conservative – parallel to the well-established leaf economic spectrum (LES). However, because of the paucity of root trait data, the complex nature of the rhizosphere, and the phylogenetic conservatism of root traits, it is unknown whether the RES hypothesis accurately describes plant nutrient and water acquisition strategies. This project utilizesplants grown in common garden conditions in the Cambridge University Botanic Garden and a meta-analysis of long-term fire manipulation experiments to examine the belowground physiological traits of fire-adapted and non-fire-adapted herbaceous species to 1) test the RES hypothesis and 2) describe the effect of fire regimes on fine root functional traits – which in turn affect carbon and nutrient cycling. A suite of morphological, chemical, and biological root traits (e.g. root diameter, specific root length, percent N, percent mycorrhizal colonization, etc.) of 50 herbaceous species were measuredand tested for phylogenetic conservatism and RES dimensionality. Fire-adapted and non-fire-adapted plants traits were compared using phylogenetic PCA techniques. Preliminary evidence suggests that phylogenetic conservatism may weaken the single-dimensionality of the RES, suggesting that there may not be a single way that plants optimize nutrient and water acquisition and storage in the complex rhizosphere; additionally, fire-adapted species are expected to be more conservative than non-fire-adapted species, which may be indicative of slower carbon cycling with increasing fire frequency and intensity.

Keywords: climate change, fire regimes, root economic spectrum, fine roots

Procedia PDF Downloads 123
6687 The First Complete Mitochondrial Genome of Melon Thrips, Thrips palmi (Thripinae: Thysanoptera): Vector for Tospoviruses

Authors: Kaomud Tyagi, Rajasree Chakraborty, Shantanu Kundu, Devkant Singha, Kailash Chandra, Vikas Kumar

Abstract:

The melon thrips, Thrips palmi is a serious pest of a wide range of agriculture crops and also act as vectors for plant viruses (genus Tospovirus, family Bunyaviridae). More molecular data on this species is required to understand the cryptic speciation and evolutionary affiliations. Mitochondrial genomes have been widely used in phylogenetic and evolutionary studies in insect. So far, mitogenomes of five thrips species (Anaphothrips obscurus, Frankliniella intonsa, Frankliniella occidentalis, Scirtothrips dorsalis and Thrips imaginis) is available in the GenBank database. In this study, we sequenced the first complete mitogenome T. palmi and compared it with available thrips mitogenomes. We assembled the mitogenome from the whole genome sequencing data generated using Illumina Hiseq2500. Annotation was performed using MITOS web-server to estimate the location of protein coding genes (PCGs), transfer RNA (tRNAs), ribosomal RNAs (rRNAs) and their secondary structures. The boundaries of PCGs and rRNAs was confirmed manually in NCBI. Phylogenetic analyses were performed using the 13 PCGs data using maximum likelihood (ML) in PAUP, and Bayesian inference (BI) in MrBayes 3.2. The complete mitogenome of T. palmi was 15,333 base pairs (bp), which was greater than the genomes of A. obscurus (14,890bp), F. intonsa (15,215 bp), F. occidentalis (14,889 bp) and S. dorsalis South Asia strain (SA1) (14,283 bp), but smaller than the genomes of T. imaginis (15,407 bp) and S. dorsalis East Asia strain (EA1) (15,343bp). Like in other thrips species, the mitochondrial genome of T. palmi was represented by 37 genes, including 13 PCGs, large and small ribosomal RNA (rrnL and rrnS) genes, 22 transfer RNA (tRNAs) genes (with one extra gene for trn-Serine) and two A+T-rich control regions (CR1 and CR2). Thirty one genes were observed on heavy (H) strand and six genes on the light (L) strand. The six tRNA genes (trnG,trnK, trnY, trnW, trnF, and trnH) were found to be conserved in all thrips species mitogenomes in their locations relative to a protein-coding or rRNA gene upstream or downstream. The gene arrangements of T. palmi is very close to T. imaginis except the rearrangements in tRNAs genes: trnR (arginine), and trnE (glutamic acid) were found to be located between cox3 and CR2 in T. imaginis which were translocated between atp6 and CR1 in T. palmi; trnL1 (Leucine) and trnS1(Serine) were located between atp6 and CR1 in T. imaginis which were translocated between cox3 and CR2 in T. palmi. The location of CR1 upstream of nad5 gene was suggested to be ancestral condition of the thrips species in subfamily Thripinae, was also observed in T. palmi. Both the Maximum likelihood (ML) and Bayesian Inference (BI) phylogenetic trees generated resulted in similar topologies. The T. palmi was clustered with T. imaginis. We concluded that more molecular data on the diverse thrips species from different hierarchical level is needed, to understand the phylogenetic and evolutionary relationships among them.

Keywords: thrips, comparative mitogenomics, gene rearrangements, phylogenetic analysis

Procedia PDF Downloads 168
6686 Phylogenetic Relationships of Common Reef Fish Species in Vietnam

Authors: Dang Thuy Binh, Truong Thi Oanh, Le Phan Khanh Hung, Luong thi Tuong Vy

Abstract:

One of the greatest environmental challenges facing Asia is the management and conservation of the marine biodiversity threaten by fisheries overexploitation, pollution, habitat destruction, and climate change. To date, a few molecular taxonomical studies has been conducted on marine fauna in Vietnam. The purpose of this study was to clarify the phylogeny of economic and ecological reef fish species in Vietnam Reef fish species covering Labridae, Scaridae, Nemipteridae, Serranidae, Acanthuridae, Lutjanidae, Lethrinidae, Mullidae, Balistidae, Pseudochromidae, Pinguipedidae, Fistulariidae, Holocentridae, Synodontidae, and Pomacentridae representing 28 genera were collected from South and Center, Vietnam. Combine with Genbank sequences, a phylogenetic tree was constructed based on 16S gene of mitochondrial DNA using maximum parsimony, maximum likelihood, and Bayesian inference approaches. The phylogram showed the well-resolved clades at genus and family level. Perciformes is the major order of reef fish species in Vietnam. The monophyly of Perciformes is not strongly supported as it was clustered in the same clade with Tetraodontiformes syngnathiformes and Beryciformes. Continue sampling of commercial fish species and classification based on morphology and genetics to build DNA barcoding of fish species in Vietnam is really necessary.

Keywords: reef fish, 16s rDNA, Vietnam, phylogeny

Procedia PDF Downloads 438
6685 A Relationship Extraction Method from Literary Fiction Considering Korean Linguistic Features

Authors: Hee-Jeong Ahn, Kee-Won Kim, Seung-Hoon Kim

Abstract:

The knowledge of the relationship between characters can help readers to understand the overall story or plot of the literary fiction. In this paper, we present a method for extracting the specific relationship between characters from a Korean literary fiction. Generally, methods for extracting relationships between characters in text are statistical or computational methods based on the sentence distance between characters without considering Korean linguistic features. Furthermore, it is difficult to extract the relationship with direction from text, such as one-sided love, because they consider only the weight of relationship, without considering the direction of the relationship. Therefore, in order to identify specific relationships between characters, we propose a statistical method considering linguistic features, such as syntactic patterns and speech verbs in Korean. The result of our method is represented by a weighted directed graph of the relationship between the characters. Furthermore, we expect that proposed method could be applied to the relationship analysis between characters of other content like movie or TV drama.

Keywords: data mining, Korean linguistic feature, literary fiction, relationship extraction

Procedia PDF Downloads 381
6684 A Case Study on Parent-Child Relationship, Attachment Styles, and Romantic Relationship Quality of Illegitimate Emerging Adults

Authors: Pierre Nicole Patriarca

Abstract:

This study examined the attachment styles, parent-child relationship, and romantic relationship quality of five illegitimate emerging adults aged 18 to 23 years old. The researcher used self-report measures, inventory of parent and peer attachment, attachment style questionnaire, and network of relationship – relationship quality version in obtaining data. A semi-structured interview was also used to acquire qualitative data about the detailed perception and experiences on the attachment styles and parent-child relationship. Common themes of each variable were identified through thematic analysis. Results showed that four out of five participants depicted positive relationship to their fathers, while all of them reported to have positive relationship to their mothers. It was also found that four participants have preoccupied attachment style, while the other one has fearful attachment style. Common themes in describing their relationship with their mother include being close, influential to participants’ life, unbounded communication, favorable reason of trusting, and sometimes being inattentive. On the other hand, having distant relationship, limited communication about romantic relationship, uninfluential to participant’s life, and favorable reason of trusting were the common themes in describing relationship with father. Lastly, less trusting, being dependent, and emphasis on valuing intimacy were the common themes in describing their style of attachment.

Keywords: illegitimate, emerging adult, attachment, parent-child relationship, relationship quality

Procedia PDF Downloads 404
6683 Molecular Characterization and Phylogenetic Analysis of Influenza a(H3N2) Virus Circulating during the 2010-2011 in Riyadh, Saudi Arabia

Authors: Ghazanfar Ali, Fahad N Almajhdi

Abstract:

This study provides data on the viral diagnosis and molecular epidemiology of influenza A(H3N2) virus isolated in Riyadh, Saudi Arabia. Nasopharyngeal aspirates from 80 clinically infected patients in the peak of the 2010-2011 winter seasons were processed for viral diagnosis by RT-PCR. Sequencing of entire HA and NA genes of representative isolates and molecular epidemiological analysis were performed. A total of 06 patients were positive for influenza A, B and respiratory syncytial viruses by RT-PCR assays; out of these only one sample was positive for influenza A(H3N2) by RT-PCR. Phylogenetic analysis of the HA and NA gene sequences showed identities higher than 99-98.8 % in both genes. They were also similar to reference isolates in HA sequences (99 % identity) and in NA sequences (99 % identity). Amino acid sequences predicted for the HA gene were highly identical to reference strains. The NA amino acid substitutions identified did not include the oseltamivir-resistant H275Y substitution. Conclusion: Viral isolation and RT-PCR together were useful for diagnosis of the influenza A (H3N2) virus. Variations in HA and NA sequences are similar to those identified in worldwide reference isolates and no drug resistance was found.

Keywords: influenza A (H3N2), genetic characterization, viral isolation, RT-PCR, Saudi Arabia

Procedia PDF Downloads 262
6682 Ranking the Elements of Relationship Market Orientation Banks (Case Study: Saderat Bank of Iran)

Authors: Sahar Jami, Iman Valizadeh

Abstract:

Today banks not only should seek for new customers but also should consider previous maintenance and retention and establish a stable relationship with them. In this term, relationship-manner marketing seeks to make, maintain, and promote the relationship between customers and other stakeholders in benefits to fulfill all involved parties. This fact is possible just by interactive transaction and promises fulfillment. According to the importance of relationship-manner marketing in banks, making context to make relationship-manner marketing has high importance. Therefore, the present study aims at exploring intention condition to relationship-manner marketing in Iran Province Iran Limited bank, and also prioritizing its variables using hierarchical analysis (AHP). There is questionnaire designed in this research to paired comparison of relationship-manner marketing elements. After distributing this questionnaire among statistical society members who are 20 of Iran Limited bank experts, data analysis has been done by Expert Choice software.

Keywords: relationship marketing, relationship market orientation, Saderat Bank of Iran, hierarchical analysis

Procedia PDF Downloads 420
6681 Computational Analyses of Persian Walnut Genetic Data: Notes on Genetic Diversity and Cultivar Phylogeny

Authors: Masoud Sheidaei, Melica Tabasi, Fahimeh Koohdar, Mona Sheidaei

Abstract:

Juglans regia L. is an economically important species of edible nuts. Iran is known as a center of origin of genetically rich walnut germplasm and expected to be found a large diversity within Iranian walnut populations. A detailed population genetic of local populations is useful for developing an optimal strategy for in situ conservation and can assist the breeders in crop improvement programs. Different phylogenetic studies have been carried out in this genus, but none has been concerned with genetic changes associated with geographical divergence and the identification of adaptive SNPs. Therefore, we carried out the present study to identify discriminating ITS nucleotides among Juglans species and also reveal association between ITS SNPs and geographical variables. We used different computations approaches like DAPC, CCA, and RDA analyses for the above-mentioned tasks. We also performed population genetics analyses for population effective size changes associated with the species expansion. The results obtained suggest that latitudinal distribution has a more profound effect on the species genetic changes. Similarly, multiple analytical approaches utilized for the identification of both discriminating DNA nucleotides/ SNPs almost produced congruent results. The SNPs with different phylogenetic importance were also identified by using a parsimony approach.

Keywords: Persian walnut, adaptive SNPs, data analyses, genetic diversity

Procedia PDF Downloads 129
6680 Validation and Interpretation about Precedence Diagram for Start to Finish Relationship by Graph Theory

Authors: Naoki Ohshima, Ken Kaminishi

Abstract:

Four types of dependencies, which are 'Finish-to-start', 'Finish-to-finish', 'Start-to-start' and 'Start-to-finish (S-F)' as logical relationship are modeled based on the definition by 'the predecessor activity is defined as an activity to come before a dependent activity in a schedule' in PMBOK. However, it is found a self-contradiction in the precedence diagram for S-F relationship by PMBOK. In this paper, author would like to validate logical relationship of S-F by Graph Theory and propose a new interpretation of the precedence diagram for S-F relationship.

Keywords: project time management, sequence activity, start-to-finish relationship, precedence diagram, PMBOK

Procedia PDF Downloads 270
6679 A Framework for Customer Knowledge Management (CKM) as a Key Role in Relationship

Authors: Mehrnoosh Askarizadeh

Abstract:

The customer’s value has become obvious for the leading companies in today’s competitive environment. Therefore they are constantly trying to improve their relationship with customers. Customer Knowledge has been recognized as a strategic resource and a key to the success of any company. Talking about the Customer Knowledge Management is closely associated with Knowledge Management and Customer Relationship Management (CRM). Recent studies conducted in the fields of Knowledge Management (KM) and Customer Relationship Management (CRM) has explained that the two approaches can have great synergies. In this paper, our aim is to provide an understanding of Customer Knowledge Management (CKM) as an integrated management approach and competence it requires. We describe CKM as an ongoing process of generating, disseminating and using customer knowledge within an organization and between an organization and its customers. In addition, we propose a comprehensive framework of CKM, the ability to integrate customer knowledge into customer relationship management processes.

Keywords: e-commerce, knowledge management (KM), customer relationship management (CRM), customer knowledge management (CKM)

Procedia PDF Downloads 557
6678 The Effects of Relationship Banking on the Financial Performance of SMEs in Kenya

Authors: Abraham Rotich

Abstract:

The purpose of this study was to determine the effects of relationship banking on the financial performance of SMEs. The paper attempted to establish the link between the constructs of relationship banking and SME performance. The study was guided by relationship lending, relationship monitoring, relationship risk sharing and bundle of products as independent variables while financial performance will be the dependent variable. The study used a quasi experimental design with population being the 620 SMEs who have a relationship banking arrangement with banks in Nairobi. The study used stratified sampling to pick a sample of 235. The population of interest will be the CEOs of the respective companies. The basis of stratification is the sectors in which the SMEs operate in. The study will use a questionnaire to collect data. The questionnaire will have both open and close ended questions. A pilot study will be conducted to test reliability and validity of questionnaire. The data was analyzed using descriptive statistics. Regression analysis was employed to test if there is a relationship between the dependent and the independent variable. The study found evidence that relationship banking positively impacts on financial performance of SMEs. Specifically, the study established that each component of relationship banking in this study i.e relationship lending, monitoring, bundle of products and risk sharing positively affects financial performance.

Keywords: relationship banking, SMEs, financial performance, entrepreneurial orientation

Procedia PDF Downloads 323
6677 Effectiveness of ISSR Technique in Revealing Genetic Diversity of Phaseolus vulgaris L. Representing Various Parts of the World

Authors: Mohamed El-Shikh

Abstract:

Phaseolus vulgaris L. is the world’s second most important bean after soybeans; used for human food and animal feed. It has generally been linked to reduced risk of cardiovascular disease, diabetes mellitus, obesity, cancer and diseases of digestive tract. The effectiveness of ISSR in achievement of the genetic diversity among 60 common bean accessions; represent various germplasms around the world was investigated. In general, the studied Phaseolus vulgaris accessions were divided into 2 major groups. All of the South-American accessions were separated into the second major group. These accessions may have different genetic features that are distinct from the rest of the accessions clustered in the major group. Asia and Europe accessions (1-20) seem to be more genetically similar (99%) to each other as they clustered in the same sub-group. The American and African varieties showed similarities as well and clustered in the same sub-tree group. In contrast, Asian and American accessions No. 22 and 23 showed a high level of genetic similarities, although these were isolated from different regions. The phylogenetic tree showed that all the Asian accessions (along with Australian No. 59 and 60) were similar except Indian and Yemen accessions No. 9 and 20. Only Netherlands accession No. 3 was different from the rest of European accessions. Morocco accession No. 52 was genetically different from the rest of the African accessions. Canadian accession No. 44 seems to be different from the other North American accessions including Guatemala, Mexico and USA.

Keywords: phylogenetic tree, Phaseolus vulgaris, ISSR technique, genetics

Procedia PDF Downloads 408
6676 Love Is Color-Blind: Perceptions among Sunway University Students toward Interracial Relationship

Authors: Azman Ramlie, Vivian Foo Jing Wen

Abstract:

As the advancement of technology has tremendously changed the way people communicate with each other, it has opened up opportunities for interracial relationships. Interracial relationship is also known as intercultural or interethnic relationships. This research topic aims to study on students’ perceptions toward interracial relationship in terms of their level of acceptance and approval. In addition, the study also focuses on determining and ranking the factors that affects students’ perceptions towards interracial relationship. This study also targets to determine the differences in students’ perceptions from business and communication major. This study uses a sample of university students from Sunway University, particularly on students in business and communication major. The study was conducted through online survey questionnaires. Results revealed that students’ level of acceptance towards interracial relationship was high. In addition, results also further indicated that family members’ support was one of the most important factors in influencing students’ perception towards interracial relationship. No diverse differences of perceptions among students in business and communication major towards interracial relationship were shown in the findings. Most of the result showed that both majors turned out to have similar perceptions towards interracial relationship. It can be said that the students’ perception towards interracial relationship did not differ from which major the students was in but their family background that would shape their perceptions.

Keywords: interracial relationship, racial, relationship, perceptions

Procedia PDF Downloads 299
6675 Systematics of Water Lilies (Genus Nymphaea L.) Using 18S rDNA Sequences

Authors: M. Nakkuntod, S. Srinarang, K.W. Hilu

Abstract:

Water lily (Nymphaea L.) is the largest genus of Nymphaeaceae. This family is composed of six genera (Nuphar, Ondinea, Euryale, Victoria, Barclaya, Nymphaea). Its members are nearly worldwide in tropical and temperate regions. The classification of some species in Nymphaea is ambiguous due to high variation in leaf and flower parts such as leaf margin, stamen appendage. Therefore, the phylogenetic relationships based on 18S rDNA were constructed to delimit this genus. DNAs of 52 specimens belonging to water lily family were extracted using modified conventional method containing cetyltrimethyl ammonium bromide (CTAB). The results showed that the amplified fragment is about 1600 base pairs in size. After analysis, the aligned sequences presented 9.36% for variable characters comprising 2.66% of parsimonious informative sites and 6.70% of singleton sites. Moreover, there are 6 regions of 1-2 base(s) for insertion/deletion. The phylogenetic trees based on maximum parsimony and maximum likelihood with high bootstrap support indicated that genus Nymphaea was a paraphyletic group because of Ondinea, Victoria and Euryale disruption. Within genus Nymphaea, subgenus Nymphaea is a basal lineage group which cooperated with Euryale and Victoria. The other four subgenera, namely Lotos, Hydrocallis, Brachyceras and Anecphya were included the same large clade which Ondinea was placed within Anecphya clade due to geographical sharing.

Keywords: nrDNA, phylogeny, taxonomy, waterlily

Procedia PDF Downloads 143
6674 Microbiological Analysis, Cytotoxic and Genotoxic Effects from Material Captured in PM2.5 and PM10 Filters Used in the Aburrá Valley Air Quality Monitoring Network (Colombia)

Authors: Carmen E. Zapata, Juan Bautista, Olga Montoya, Claudia Moreno, Marisol Suarez, Alejandra Betancur, Duvan Nanclares, Natalia A. Cano

Abstract:

This study aims to evaluate the diversity of microorganisms in filters PM2.5 and PM10; and determine the genotoxic and cytotoxic activity of the complex mixture present in PM2.5 filters used in the Aburrá Valley Air Quality Monitoring Network (Colombia). The research results indicate that particulate matter PM2.5 of different monitoring stations are bacteria; however, this study of detection of bacteria and their phylogenetic relationship is not complete evidence to connect the microorganisms with pathogenic or degrading activities of compounds present in the air. Additionally, it was demonstrated the damage induced by the particulate material in the cell membrane, lysosomal and endosomal membrane and in the mitochondrial metabolism; this damage was independent of the PM2.5 concentrations in almost all the cases.

Keywords: cytotoxic, genotoxic, microbiological analysis, PM10, PM2.5

Procedia PDF Downloads 345
6673 Genomic Characterisation of Equine Sarcoid-derived Bovine Papillomavirus Type 1 and 2 Using Nanopore-Based Sequencing

Authors: Lien Gysens, Bert Vanmechelen, Maarten Haspeslagh, Piet Maes, Ann Martens

Abstract:

Bovine papillomavirus (BPV) types 1 and 2 play a central role in the etiology of the most common neoplasm in horses, the equine sarcoid. The unknown mechanism behind the unique variety in a clinical presentation on the one hand and the host-dependent clinical outcome of BPV-1 infection, on the other hand, indicate the involvement of additional factors. Earlier studies have reported the potential functional significance of intratypic sequence variants, along with the existence of sarcoid-sourced BPV variants. Therefore, intratypic sequence variation seems to be an important emerging viral factor. This study aimed to give a broad insight in sarcoid-sourced BPV variation and explore its potential association with disease presentation. In order to do this, a nanopore sequencing approach was successfully optimized for screening a wide spectrum of clinical samples. Specimens of each tumour were initially screened for BPV-1/-2 by quantitative real-time PCR. A custom-designed primer set was used on BPV-positive samples to amplify the complete viral genome in two multiplex PCR reactions, resulting in a set of overlapping amplicons. For phylogenetic analysis, separate alignments were made of all available complete genome sequences for BPV-1/-2. The resulting alignments were used to infer Bayesian phylogenetic trees. We found substantial genetic variation among sarcoid-derived BPV-1, although this variation could not be linked to disease severity. Several of the BPV-1 genomes had multiple major deletions. Remarkably, the majority of the cluster within the region coding for late viral genes. Together with the extensiveness (up to 603 nucleotides) of the described deletions, this suggests an altered function of L1/L2 in disease pathogenesis. By generating a significant amount of complete-length BPV genomes, we succeeded in introducing next-generation sequencing into veterinary research focusing on the equine sarcoid, thus facilitating the first report of both nanopore-based sequencing of complete sarcoid-sourced BPV-1/-2 and the simultaneous nanopore sequencing of multiple complete genomes originating from a single clinical sample.

Keywords: Bovine papillomavirus, equine sarcoid, horse, nanopore sequencing, phylogenetic analysis

Procedia PDF Downloads 178