Presentation Title

The Biochemistry and Bioinformatics of Selected Ocular Proteins from Bos taurus

Faculty Mentor

Robert Richards, PhD

Start Date

18-11-2017 10:00 AM

End Date

18-11-2017 11:00 AM

Location

BSC-Ursa Minor 52

Session

Poster 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Infectious bovine keratoconjunctivitis commonly referred to, as “pinkeye” in cattle, is an ocular disease that is endemic in cattle herds worldwide. Our previous study determined the bacteria present in each condition whereas this study focused on identifying genes known to cause infection in the healthy versus infected cattle and to learn more about the virulence mechanisms used by pathogenic bacteria. This study involves the verification and use of MG-RAST, a web application that analyses metagenomes. Two methods were used to establish the reliability of the data produced by MG-RAST. The first technique exploited a sample that contained only E. coli as the sole bacteria identified. By utilizing the KEGG database it was possible to identify more than 75% of the putative proteins included in a number of known pathways. The second technique utilized the glycolysis/gluconeogenesis pathway in KEGG. Comparing all the samples to the pathway more than 85% of the putative proteins were identified (n=18, SD = 1.8). These two techniques suggested that MG-RAST identified a majority of the proteins present. The second objective was to identify functional differences in healthy versus infected samples to investigate how bacteria infect calves. Commonly known virulent factors and pathways were considered. Our investigation showed a significant lack of genes from the type III secretion system and the two-component system genes AgrA, SaeS and SaeR in healthy samples even though some contained known pathogens. These missing virulent factors, required for pathogenicity, suggest a possible explanation of the noted avirulence. This study of virulent factors derived from functional metadata can be used to gain a better understanding of the biological interactions and virulent mechanisms in a mixed environment of organisms.

Summary of research results to be presented

  • Two methods were used in tandem to establish the reliability of the data produced by MG-RAST, an open source web application that proposes functional analysis of metagenomes.
  • The first technique exploited a sample containing only Escherichia coli as the sole bacteria identified. Then by utilizing KEGG database it was possible to identify more than 75% of the putative proteins included in the pathways of each functional category suggesting that the software is proficient in identifying majority of the functional genes involved in these pathways (n=10). The second technique utilized the functional data of all the samples identified in the glycolysis/gluconeogenesis pathway from KEGG. More than 85% of the genes were identified in all the samples with very little differences between samples (n=18, S.D. = 1.8) suggesting that MG-RAST identified a majority of the genes present.
  • The second objective of the project was to identify functional differences in the healthy versus infected samples to investigate how the bacteria infect the calves. Commonly known virulent factors were considered.
  • The type III secretion system is the most critical of the virulent secretion systems due to its “injector” component of the protein and was found in 5 out of the 8 infected samples. The T3SS was found to be absent in all the healthy samples other than two. Interestingly, these two healthy samples contained bacteria that had most of the T3SS genes but lacked the genes for the “injector” portion of the protein, suggesting a reason for the virulent condition.

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Nov 18th, 10:00 AM Nov 18th, 11:00 AM

The Biochemistry and Bioinformatics of Selected Ocular Proteins from Bos taurus

BSC-Ursa Minor 52

Infectious bovine keratoconjunctivitis commonly referred to, as “pinkeye” in cattle, is an ocular disease that is endemic in cattle herds worldwide. Our previous study determined the bacteria present in each condition whereas this study focused on identifying genes known to cause infection in the healthy versus infected cattle and to learn more about the virulence mechanisms used by pathogenic bacteria. This study involves the verification and use of MG-RAST, a web application that analyses metagenomes. Two methods were used to establish the reliability of the data produced by MG-RAST. The first technique exploited a sample that contained only E. coli as the sole bacteria identified. By utilizing the KEGG database it was possible to identify more than 75% of the putative proteins included in a number of known pathways. The second technique utilized the glycolysis/gluconeogenesis pathway in KEGG. Comparing all the samples to the pathway more than 85% of the putative proteins were identified (n=18, SD = 1.8). These two techniques suggested that MG-RAST identified a majority of the proteins present. The second objective was to identify functional differences in healthy versus infected samples to investigate how bacteria infect calves. Commonly known virulent factors and pathways were considered. Our investigation showed a significant lack of genes from the type III secretion system and the two-component system genes AgrA, SaeS and SaeR in healthy samples even though some contained known pathogens. These missing virulent factors, required for pathogenicity, suggest a possible explanation of the noted avirulence. This study of virulent factors derived from functional metadata can be used to gain a better understanding of the biological interactions and virulent mechanisms in a mixed environment of organisms.