A. Informații generale
Titlul proiectului:
Identificarea mecanismelor complexe de adaptare ale micobacteriilor la medii sulfidice extreme cu interfețe redox gaz-gaz
Identification of mycobacteria's complex mechanisms of adaptation to extreme sulfidic environments with gas-gas redox interfaces
Acronimul proiectului: Mycored
Domeniul științific: 4. Sănatate
Fonduri solicitate de la bugetul subprogramului: 38.475,00 lei
Rezumat:
Biotehnologiile microbiene joacă un rol crucial în dezvoltarea de noi antibiotice, în promovarea medicinei personalizate și a terapiilor țintite prin valorificarea înțelegerii interacțiunilor gazdă-microbiom. Obiectivul principal al acestui proiect este identificarea mecanismelor complexe de adaptare a micobacteriilor la medii sulfidice extreme cu interfețe redox gaz-gaz. Studiul este realizat pe o comunitate microbiană extremofilă care conține micobacterii, situată într-o peșteră sulfuroasă din România. Condițiile de viață extrem de particulare întâlnite în acest mediu extrem ne oferă oportunitatea unică de a studia influența factorilor de mediu asupra expresiei genetice în micobacterii. Comparațiile cu agenții patogeni înrudiți ne vor oferi informații extrem de utile despre exprimarea anumitor caractere de patogenitate, precum și despre mecanismele de adaptare a micobacteriilor la acest tip de mediu. Astfel, am putea vizualiza proiectarea unor posibile noi terapii. Vizitele bilaterale din cadrul proiectului de mobilitate Mycored vor consolida relația dintre cei 2 parteneri și vor duce la realizarea unui amplu proiect de cercetare finanțat de Comunitatea Europeană (CE).
Directorul de proiect România:
GHERMAN Vasile Daniel (https://brainmap.ro/)
Instituția gazdă(coordonatoare) a proiectului din România:
Universitatea Politehnica Timisoara
Coordonator de proiect Belgia
Jean-François FLOT
Instituția gazdă(coordonatoare) Belgia:
Université libre de Bruxelles (ULB)
Pagină web: https://ebe.ulb.be/ebe/Flot.html
Cuvinte cheie: micobacterii, biotehnologii microbiene, expresie genetica, caracter de patogenitate
B. Description of the bilateral cooperation project
B1. Partners involved in the project.
- 2 partners are involved in this project: P1. The Microbiological Analysis and Microbiological Risk Management Laboratory of the Politehnica University of Timisoara (UPT), Romania, coordinated by Dr. Vasile Daniel GHERMAN and P2. The Ecological and Evolutionary Genomics research team with the research unit Evolutionary Biology & Ecology of the Université libre de Bruxelles (ULB), headed by Dr. Jean-François FLOT.
The 2 partners involved in the project started together, 3 years ago, the analysis of microbial communities from different extreme sulfidic environments with possible implications in medical biotechnologies. During this period, the 2 partners proved a great complementarity and compatibility. Thus P1 (UPT) managed the collection of samples, isolation, cultivation on growth media and obtaining pure cultures from the studied microbial consortia. P2 (ULB) managed the metagenomic analysis of the microbial communities and used the isolates obtained by P1 (UPT) for sequencing and complete genomic mapping. This collaboration resulted in the joint publication in November 2022 of an article in the prestigious journal Nature Microbiology (DOI10.1038/s41564-022-01252-3).
B2. Activities carried out within the project.
Scientific relevance/Motivation of the proposed project;
The exploration of microbial communities present in some environments considered extreme can provide solutions with implications in medicine and microbial biotechnologies. 3 years ago the two partners of the Mycored project started investigating the microbial communities present in a unique sulphidic cavity - Sulfur Cave (Puturosu Mountain, Romania). The uniqueness of this ecosystem consists in the fact that the microbial community is arranged in the gas chemocline. The upper portion of the cave is filled with air that floats above the heavier volcanic gases, while the lower section is filled with mofettic CO2, CH4, and H2S. The cave wall is covered in S° and H2SO4 (from sulfur-oxidation) at and below the CO2-H2S:O2 gas/gas contact. The pH is less than one on the cave wall close to the contact, and peculiar microbiological biofilms on the surface of the rock were found. The only source of liquid water in this system is the condensation of water vapor onto the cave walls. Ferroplasmaceae, Acidithiobacillus, and mycobacteria are the most prevalent taxa in these microbial biofilms (https://doi.org/10.5038/1827-806X.47.2.2164).
The main motivation of this project proposal starts from the fact that the study of mycobacteria from this unique microbial community can provide very interesting perspectives for microbial biotechnologies.
Specific objectives of the Mycored project:
O1. Preparing of pure cultures of microorganisms to be sent to two public microorganism repositories;
O2. Depositing pure cultures of microorganisms;
O3. Description and characterization of the new species of microorganisms;
O4. Investigation of how environmental factors affect mycobacteria's genomic expression;
O5. Dissemination of results.
Work plan
The Mycored project includes 3 work packages. Both partners of the Mycored project will be involved in the 3 work packages.
WP1. The preparation and deposit of pure cultures of the new species isolated from the biofilm of Sulfur Cave in the international collections of microorganisms. Their genetic mapping and species description. Months 1-6.
A.1.1 Preparation of pure cultures of microorganism to be sent to two public microorganism repositories, one in Belgium and one in Switzerland;
The discovery and study of new taxa in the field of Prokaryota can be of particular importance in biotechnologies and in medicine in general. In this way, research is crucial to expanding our understanding of microbial diversity, which affects a wide range of disciplines, including ecology, industry, environmental science and medicine. The number of NTM patients identified annually worldwide has steadily increased over the last 20 years (Ratnatunga et al., 2020). Improved understanding of NTM and more accessible diagnostic tools are likely contributing causes to this increase, while the precise cause is yet unknown. The variety and ubiquity of NTM in the environment are yet unknown to us, though. To increase our knowledge and the tests that may be used to accurately diagnose NTM infections, it is imperative that we continue to study and characterize NTM.
From a morphological point of view, the colonies on both media tested were relatively round, moist, raised with irregular edges, mucous appearance and consistency, orange in color (Fig. 1). The colonies appeared on the culture medium after 3 weeks, have a yellow-orange color, variable widths of raised irregular edges and a mucous appearance. Colonies were scotochromogenic, i.e., orange pigmentation was produced regardless of light exposure or incubation temperature (Fig. 1). The isolate grew well at 30°C and 37°C, but no growth was observed at 42°C. Microscopic examination after acid-fast Ziehl-Neelsen staining confirmed the presence of AFB that were 1–3 μm in length.
Figure 1
Colony morphology of M. balvany after 4 weeks incubation at 33°C on Löwenstein Jensen plate.
Figure 2 TEM images of Mycobacterium balvany
A.1.2 Creation of submission documents and submission of microorganisms in the mentioned public cultures;
A.1.3 Genetic mapping and species description.
From a phylogenetic point of view, M. balvany is close to M. bohemicum. M. balvany (M. Mag 2 or Crude 2) and M. bohemicum are separated from a branch with Mycobacterium saskatchewanense. Both M. bohemicum and M. saskatchewanense have pathogenic characteristics.
WP2. The study of the interspecific relationships between consortium’s microorganisms. Study of genetic expression and comparisons with related pathogenic microorganisms. Months 7-16.
A.2.1 Biofilm sampling from Sulfur caves and cultivation of microorganisms on different types of growth media; A.2.2 Study of genetic expression; A.2.3 Comparisons with related pathogenic microorganisms.
WP3. Project Management and dissemination. Months 1-16.
A.3.1 Project Management. The management team P1 (UPT) and P2 (ULB) has the role of assuring the efficient use of human, material and financial resources in order to have a solid project progress; A.3.2 Dissemination of results. During the project, 2 ISI rated articles will be published.