The Use of Bioinformatic Tools in Symbiosis and Co-Evolution Studies

Through millions of years, the multicellular organisms have coexisted and coevolved with the surrounding microorganisms, in an almost symbiotic relationship forming a complex entity known as holobiont. The composition and functions of these microbial communities were limited during many years to only a mere fraction, due to the use of culture-based techniques. The advent of molecular-based techniques allowed the identification of uncultured organisms in a culture-free manner. In recent years, the development of next generation sequencing techniques have allowed the high-throughput study of microbial communities allowing the identification and classification of otherwise uncultured microorganisms in a given environment, tissue or host through metagenomics. The next generation sequencing techniques have been used in the functional study of microbial assemblages and were able to identify the role of the microorganisms in biogeochemical cycles, pathogenic processes, metabolism and development, through metatranscriptomics. Taken together, the next generation sequencing based-studies have shown the existence of a complex metabolic network in different hosts and environments, with the microbial communities. This chapter will focus in different available bioinformatic tools that are suitable to study symbiosis and coevolution processes in a given sample

DNA microarrays to identify etiological agents, as sensors of environmental wellbeing

Background: The epidemiologic transition in Mexico has generated a change of paradigm in public health. Morbidity is characterized by infectious diseases and the mortality is due to chronic degenerative diseases. The three most important infectious diseases in the country are: respiratory infections, diarrhea, and urinary tract infections.Method: The objective of this work was to build a tool to monitor the presence of health risks in the environment in a timely manner and to demonstrate its application in different sicknesses, especially those that are water related. In this study, we analyzed water samples from five cenotes with high tourist flow in the State of Yucatan. We developed a DNA microarray for the adequate and prompt detection of viruses, bacteria, fungi, and parasites. This microarray could be used in samples of different origin including air, water (fresh, brackish and saltwater), food, inert surfaces or wounds. Clinically, it would allow prompt and precise detection of etiological agents of infectious diseases to prevent outbreaks. It would also be useful for the identification of those agents that cannot be detected in our laboratories with the traditional methods. It includes 38,000 probes that detect 252 etiological agents of diseases in humans and antimicrobial resistance genes. Results from DNA samples can be obtained in 24 h, which would be difficult or impossible using other technologies.Results: The results are readily available within 24 h. Samples from five cenotes (sinkholes) with high flow of people, were analyzed with the microarray. The water samples analyzed detected 228 different bacteria, viruses, fungi, and protozoa. They are amongst the most important etiological agents for infectious diseases in Mexico.Conclusions: The microarray provides the opportunity for precise and early detection of various infectious agents in individuals, hospitals and natural environments. This could help reduce the global burden of diseases, the severity of outbreaks, and reduce antibiotic resistance

RNA-Seq Analysis on the Microbiota Associated with the White Shrimp (Litopenaeus vannamei) in Different Stages of Development

White leg shrimp (Litopenaeus vannamei) is a widely cultured species along the Pacific coast and is one of the most important crustaceans in world aquaculture. The microbiome composition of L. vannamei has been previously studied in different developmental stages, but there is limited information regarding the functional role of the microbiome during the development of L. vannamei. In this study the metatranscriptome in different developmental stages of L. vannamei (larvae, juvenile and adult) were generated using next generation sequencing techniques. The bacterial phyla found throughout all the stages of development belonged to the Proteobacteria, Firmicutes and Actinobacteria, these bacterial phyla are present in the digestive tract and are capable of producing several hydrolytic enzymes, which agrees with high representation of the primary metabolism and energy production, in both host and the microbiome. In this sense, functional changes were observed as the development progressed, in both host and the microbiome, in stages of larvae the most represented metabolic functions were associated with biomass production; while in juvenile and adult stages a higher proportion of metabolic functions associated to biotic and abiotic stress in L. vannamei and the microbiome were shown. This study provides evidence of the interaction of the microbiome with L. vannamei, and how the stage of development and the culture conditions of this species influences the gene expression and the microbiome composition, which suggests a complex metabolic network present throughout the life cycle of L. vannamei

RNA-Seq Analysis on the Microbiota Associated with the White Shrimp (<i>Litopenaeus vannamei</i>) in Different Stages of Development

White leg shrimp (Litopenaeus vannamei) is a widely cultured species along the Pacific coast and is one of the most important crustaceans in world aquaculture. The microbiome composition of L. vannamei has been previously studied in different developmental stages, but there is limited information regarding the functional role of the microbiome during the development of L. vannamei. In this study the metatranscriptome in different developmental stages of L. vannamei (larvae, juvenile and adult) were generated using next generation sequencing techniques. The bacterial phyla found throughout all the stages of development belonged to the Proteobacteria, Firmicutes and Actinobacteria, these bacterial phyla are present in the digestive tract and are capable of producing several hydrolytic enzymes, which agrees with high representation of the primary metabolism and energy production, in both host and the microbiome. In this sense, functional changes were observed as the development progressed, in both host and the microbiome, in stages of larvae the most represented metabolic functions were associated with biomass production; while in juvenile and adult stages a higher proportion of metabolic functions associated to biotic and abiotic stress in L. vannamei and the microbiome were shown. This study provides evidence of the interaction of the microbiome with L. vannamei, and how the stage of development and the culture conditions of this species influences the gene expression and the microbiome composition, which suggests a complex metabolic network present throughout the life cycle of L. vannamei