Bioluminescence-based Reporter Bacteriophages: New Technology for Monitoring Microbes and Diseases

Bacteria and bacteriophage. Photo by Oxford (Article by Pete Wilton)

In recent years, the development of bioluminescence-based reporter bacteriophages has revolutionized the monitoring of microbes and their diseases. These reporter bacteriophages are genetically engineered to carry a gene that will be integrated into the host genome which will at one point reflect the fluorescence signals. Bioluminescence reporter bacteriophages have high specificity and sensitivity, making them useful tools for studying microbes and their diseases. They are also useful research tools that enable researchers to monitor the efficacy of bacteriophage/drug treatments, the spread of infection, the success of phage delivery, and the activity of phage-resistant bacteria. In this article, we provide an overview of bioluminescence reporter bacteriophages, describe their applications and advantages, and discuss how you can use them in your own research.

What is Bioluminescence?

Bioluminescence is the production and emission of light by living organisms. This light is produced by the living cell through an enzyme called luciferase. This enzyme catalyzes the transformation of the light-emitting substrate, luciferin, into oxyluciferin, which can then be converted to light by light-emitting complexes. The light-emitting complexes are also called photoproteins. The overall process of bioluminescence is called photoautotrophy. In photoautotrophs, light energy from the sun is used to synthesize chemical energy and regulate cell metabolism, in order to provide energy and materials to the organism. In photoheterotrophs, light energy is used to regulate metabolism and produce energy by photosynthesis. In photoautotrophs, the production of light energy is coupled with the production of essential chemicals in the form of macromolecules such as carbohydrates, lipids, proteins, and nucleic acids. In photoheterotrophs, the production of light energy is not coupled with the production of essential chemicals. Instead, the photosynthesis process uses the energy from light to synthesize complex organic molecules such as carbohydrates, lipids, and proteins. In addition to producing light energy and essential chemicals, photosynthesis requires the production of a supply of carbon. The production of this carbon supply is called photosynthetic carbon fixation. This process of photosynthetic carbon fixation is what produces bioluminescence in many types of organisms.

Bioluminescence Reporter Bacteriophages: The Basics

Bioluminescent reporter bacteriophages are genetically modified viruses that infect their host cells with high specificity and transduce a heterologous luciferase gene, the activity of which can be detected with high sensitivity to indicate the presence of viable target cells. Bacteriophages (phages) evolved as natural predators of bacteria to bind their hosts with unparalleled specificity and to rapidly deliver and replicate their viral genome. Not surprisingly, phages and phage-encoded proteins have been used to create a diverse set of diagnostic assays, many of which outperform traditional culture-based and molecular detection methods. While intact phages or phage-encoded affinity proteins can be used to capture bacteria, most phage-inspired detection systems rely on viral genome delivery and amplification: suitable phages are genetically reprogrammed to deliver heterologous reporter genes, the activity of which is typically detected via enzyme action on substrate conversion to indicate the presence of a viable host cell. Infection with this kind of engineered reporter phages usually results in a rapid burst of reporter protein production, allowing for highly sensitive detection.

Uses of Reporter bacteriophages

Bacterial pathogens in clinical samples can be detected quickly and reliably.

Assessing contaminations in samples (e.g water and food samples)
Monitoring spread of infection or contamination

Advantage of using reporter bacteriophages

They are specific

They are reliable
They are fast (act quickly)
Has potential for automation

Easy to make them reproducible and user friendly

How to use Reporter Bacteriophages in your research

Before discussing the applications of bioluminescence reporter bacteriophages in biology, it is important to consider how they are used in research. Although the majority of bioluminescence reporter bacteriophages are used as research tools, there are a few specific applications of reporter phages in healthcare and agriculture that researchers can also use them for.

Healthcare

Healthcare researchers use bioluminescence reporter phages for two main applications: monitoring the efficacy of treatments and tracking the spread of infection. The first application involves using bioluminescence reporter phages to monitor the efficacy of the anti-microbial agents in controlling the growth of disease-causing bacteria. Because the efficacy of antibiotic treatments can vary depending on the species of bacteria, monitoring the growth of bacteria in the presence of antibiotic compounds is highly important to researchers in the field of antibiotic design. For example, if researchers want to know how well an antibiotic works against bacteria in the Staphylococcus aureus group, they can genetically engineer a bioluminescence reporter bacteriophage to express luciferase in the S. aureus group and infect bacteria. When the bacteria are infected with the bioluminescence reporter phage, they will express the luciferase enzyme, which indicates that the antibiotic administered to the bacteria is having an effect on them and can be used to design better drugs.

Agriculture

Bioluminescence reporter bacteriophages are also used to track the spread of infections in livestock. As most animals are susceptible to infections from bacteria, researchers use bioluminescence reporter phages to monitor the spread of infectious agents within animals. This can be used to track the success of vaccination programs and the efficacy of anti-bacterial compounds and vaccination strategies. For example, a cattle ranch might want to know how effective a vaccination program is at controlling the spread of infectious bovine pneumonia (IBPP) in their animals. After vaccinating their cattle, researchers can use a bioluminescence reporter phage to track the spread of infection within their animals. When a bioluminescence reporter phage infects a susceptible bovine, they will express the enzymes luciferase and green fluorescent protein, which indicates that they have been infected with IBPP and will give off light signals until they die of infection. From this, researchers can determine the success of their vaccination program. Another application of bioluminescence reporter bacteriophages in agriculture is in aquaculture where a farmer can easily monitor the spread of infections from farm to farm until its managed.

NB:This technology (Bioluminescence-based Reporter Bacteriophages) can be used in a variety of other fields (food processing industries e.t.c) that involve the detection of bacteria.

Conclusion

Although bioluminescence reporter bacteriophages have been used mainly as research tools, this technology is at an early stage of development and has a wide range of applications in medicine and agriculture. They have a wide range of uses in both research and healthcare, including monitoring the efficacy of antibiotics and the spread of infection within organisms. They also have far-reaching potential in agriculture and environmental monitoring. The development of this technology has led to the discovery of new ways to track microbes and the spread of infection, and provides new tools for scientists to study how microbes interact with their environments.

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