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An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines
Published on: October 11, 2022
[Emerging rickettsioses]
1Dipartimento di Patologia Animale Veterinaria, Igiene e Sanità Pubblica Veterinaria, Università degli Studi di Milano, Via Celoria, 22, 20133 Milano, Italy.
Abstract:
Ticks are known to carry and transmit a number of microbial agents that cause diseases in humans and animals. Among these are members of the order Rickettsiales (alpha-proteobacteria), which include the genera Rickettsia and Ehrlichia. The most common and well-known Rickettsial human disease in Europe is Mediterranean Spotted Fever (MSF), caused by Rickettsia conorii. In recent years, a number of new Rickettsia species have been discovered in Europe, some of which have been shown to be pathogenic to humans. These discoveries have been facilitated by use of sequence-based molecular identification techniques. In Italy, it is generally believed that R. conorii is the only Rickettsia species present, and clinical tests for MSF rely on antigens raised against this bacterium. We are currently undertaking a molecular screening study of Rickettsiales-bacteria in ticks from various regions of Italy, to check for the potential presence of species from this order recently discovered in other parts of Europe. So far, we have identified a number of additional species in ticks collected from northern, central and southern regions. These include the known pathogens R. helvetica and R. slovaca as well as two species which may or may not be of medical relevance: R. monacensis and R. sp. IRS4. As a part of this survey, we have identified a novel alphaproteobacterium from the medically important tick Ixodes ricinus. This bacterium, tentatively named IricES1, has the unusual property of existing within the mitochondria, as well as the cytoplasm, of ovarian cells. To our knowledge, this is the only known example of a bacterium that is able to enter the mitochondria of animals. Our recently published electron microscopic data indicates that the bacterium enters mitochondria between the inner and outer membranes, and then proceeds to consume the inner mitochondrial matrix. We will present further data on this bacterium, including: 1) its phylogenetic position based on various molecular sequences, 2) its localization within the tick based on in situ hybridization; 3) its distribution among tick populations in Europe; 4) preliminary data on attempts at culturing this bacterium in a variety of cell types. Possible interactions between the bacterium and its host will be discussed. Ticks are known to carry and transmit a number of microbial agents that cause diseases in humans and animals. Among these are members of the order Rickettsiales (alpha-proteobacteria), which include the genera Rickettsia and Ehrlichia. The most common and well-known Rickettsial human disease in Europe is Mediterranean Spotted Fever (MSF), caused by Rickettsia conorii. In recent years, a number of new Rickettsia species have been discovered in Europe, some of which have been shown to be pathogenic to humans. These discoveries have been facilitated by use of sequence-based molecular identification techniques. In Italy, it is generally believed that R. conorii is the only Rickettsia species present, and clinical tests for MSF rely on antigens raised against this bacterium. We are currently undertaking a molecular screening study of Rickettsiales-bacteria in ticks from various regions of Italy, to check for the potential presence of species from this order recently discovered in other parts of Europe. So far, we have identified a number of additional species in ticks collected from northern, central and southern regions. These include the known pathogens R. helvetica and R. slovaca as well as two species which may or may not be of medical relevance: R. monacensis and R. sp. IRS4. As a part of this survey, we have identified a novel alphaproteobacterium from the medically important tick Ixodes ricinus. This bacterium, tentatively named IricES1, has the unusual property of existing within the mitochondria, as well as the cytoplasm, of ovarian cells. To our knowledge, this is the only known example of a bacterium that is able to enter the mitochondria of animals. Our recently published electron microscopic data indicates that the bacterium enters mitochondria between the inner and outer membranes, and then proceeds to consume the inner mitochondrial matrix. We will present further data on this bacterium, including: 1) its phylogenetic position based on various molecular sequences, 2) its localization within the tick based on in situ hybridization; 3) its distribution among tick populations in Europe; 4) preliminary data on attempts at culturing this bacterium in a variety of cell types. Possible interactions between the bacterium and its host will be discussed.
Insights
Ticks can carry various Rickettsiales bacteria, including new species found in Italy. Researchers discovered a novel bacterium, IricES1, within the mitochondria of Ixodes ricinus ticks, a unique finding with potential health implications.
Area of Science:
- Microbiology
- Molecular Biology
- Parasitology
Context:
- Ticks transmit numerous microbial agents, including Rickettsiales (Rickettsia, Ehrlichia).
- Mediterranean Spotted Fever (MSF), caused by Rickettsia conorii, is a common Rickettsial disease in Europe.
- Recent discoveries of novel Rickettsia species in Europe highlight the need for ongoing surveillance.
Purpose:
- To conduct a molecular screening of Rickettsiales in Italian ticks.
- To identify Rickettsia species beyond R. conorii in Italy.
- To characterize a novel alphaproteobacterium found within Ixodes ricinus ticks.
Summary:
- Molecular screening of ticks across Italy revealed diverse Rickettsiales, including known pathogens (R. helvetica, R. slovaca) and potentially relevant species (R. monacensis, R. sp. IRS4).
- A novel bacterium, provisionally named IricES1, was identified in Ixodes ricinus ticks.
- IricES1 uniquely resides within the cytoplasm and mitochondria of tick ovarian cells, consuming the inner mitochondrial matrix.
Impact:
- Expands the known diversity of Rickettsiales in Italian ticks.
- Identifies potential novel tick-borne pathogens.
- Describes a unique bacterium with an unprecedented ability to invade animal mitochondria, opening new avenues for research into host-microbe interactions and mitochondrial biology.
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