NAME – SAURABH MANDAL ROLL NO.- RP7002AO1 SECTIONP7002 COURSE CODE- BTY001A COURSE NAME- GENERAL SEMINAR
->Phage therapy ->What is the need? ->Bacteriophages – raw material of the therapy ->History ->How it happen? ->Structure of Bacteriophage ->Phage life cycle: Lytic vs Lysogenic ->Adsorption by Lytic Bacteriophage ->Penetration ->Early Replication ->Phage Release ->Conventional Bacteriophage Therapy in humans ->Advantages ->Limitations of phage therapy ->Combating the limitations
Phages therapy were being used therapeutically to treat bacterial infections that do not respond to conventional antibiotics.
Phage therapy is one of the viable alternatives to antibiotics An important benefit of phage therapy is that bacteriophages can be much more specific than more common drugs and thus harmless to not only the host organism (human, animal or plant) and to ecology.
What is the need?
There is an urgent need to develop new classes of antibiotics to tackle the increase in resistance in many common bacterial pathogens. Pathogens such as Staphylococcus aureus, Streptococcus pneumoniae and Enterococcus faecalis, which are each capable of causing severe and even fatal infections , have become increasingly resistant to multiple antibiotics.
Bacteriophages – raw material of the therapy
Bacterial viruses(kill bacteria)-very specific host range, short replication time, self amplifying bacteriophages use a syringe-like motion to inject their genetic material into the cell. After making contact with the appropriate receptor, the tail fibers bring the base plate closer to the surface of the cell. Once attached completely, the tail contracts, injecting genetic material through the bacterial membrane. Natural in soil and foods
The genetic material of Bacteriophages can be ssRNA, dsRNA, ssDNA, or dsDNA. 1966.....Norton D. Zinder discovers RNA bacteriophages.
Since ancient times, there have been documented reports of river water having the ability to cure infectious diseases…
In 1896, Ernest Hanbury Hankin reported that something in the waters of the Ganges and Jumna rivers in India had marked antibacterial action against cholera. French-Canadian microbiologist Félix d'Hérelle, working at the Pasteur Institute in Paris, in 1917 discovered the bacteriophage. D'Hérelle called the virus a bacteriophage or bacteria-eater.
Following the discovery of bacteriophages by Frederick Twort and Felix d'Hérelle in 1915 and 1917, phage therapy was immediately recognized by many to be a key way forward for the eradication of bacterial infections Isolated from Western advances in antibiotic production in the 1940s, Russian scientists continued to develop already successful phage therapy to treat the wounds of soldiers in field hospitals Bacteriophages or phages are bacterial viruses that invade bacterial cells and, in the case of lytic phages, disrupt bacterial metabolism and cause the bacterium to lyse.
How it happen?
Collecting local samples of water likely to contain high quantities of bacteria and bacteriophages, for example effluent outlets, sewage and other sources. The samples are taken and applied to the bacteria that are to be destroyed which have been cultured on growth medium. Phages are "bacterium-specific" and it is therefore necessary in many cases to take a swab from the patient and culture it prior to treatment. Occasionally, isolation of therapeutic phages can require a few months to complete, but clinics generally keep supplies of phage cocktails for the most common bacterial strains in a geographical area.
Phages in practice are applied orally, topically on infected wounds or spread onto surfaces, or used during surgical procedures. Injection is rarely used, avoiding any risks of trace chemical contaminants that may be present from the bacteria amplification stage and recognizing that the immune system naturally fights against viruses introduced into the bloodstream or lymphatic system.
from the greek phagein, meaning "to eat“ Eaters or destroyers of bacteria
First described in 1915
SEM of Phage
Structure of Bacteriophage
•Phage head: composed of coat protein and genome in the core •Genome: DNA codes for enzymes and proteins necessary to replicate more viruses •Tail Sheath: DNA travels from head to bacteria through sheath •Tail fiber: helps anchor the phage on the cell membrane
Phage life cycle: Lytic vs Lysogenic
Phage replicates by lytic life cycle Non-integration of phage genetic material Phage lyse host bacterium lytic or virulent phage
Phage replicates by lysogenic life cycle Integration of phage genetic material temperate phages (prophages) generally larger than lytic phages
Adsorption by Lytic Bacteriophage
The bacteriophage binds to specific receptors on the bacterial cell wall.
Tail conformation changes/contracts central core penetrates cell wall
The bacteriophage injects its genome into the bacterium's cytoplasm
-Phage-coded enzymes shut down host‟s DNA,RNA,protein synthesis
-Early function inovolve the takeover of the host cell and the synthesis o early viral mRNA
-Late functions include the subsequent synthesis of other proteins and assembly of the nucleocapsid.
-Replication phage DNA protected from host restriction endonucleaes
A bacteriophage-coded enzyme break down the peptidoglycan in the bacterial cell wall causing osmotic lysis.
Conventional Bacteriophage Therapy in humans
Biomedical technology today is very different from what it was in the early days of phage therapy research In early days bacteriophage therapy was used by making bacteriophage preparation and are effective against P. aeruginosa, E.coli, S.aureus, Streptococcus and proteus The first reviwed report of the therapeutic efficacy of PhagoBioDerm
Treated with phage impregnated pad
Improvement in wound healing
FOR THE TREATMENT OF A VARIETY OF BACTERIAL INFECTIONS INCLUDING: laryngitis, skin infections, dysentery, conjunctivitis, gingivitis, sinusitis, urinary tract infections and intestinal infections, burns, boils, poly-microbial biofilms on chronic wounds, ulcers and infected surgical sites REDUCE THE USE OF ANTIBIOTIC-The problem that overuse of antibiotics creates is the emergency of antibioticresistant strains of certain diseases. There are several types of disease that are becoming resistant to various antibiotic drugs, making them more difficult to treat successfully especially in case of bacterial infection.
Minimises any response from gut flora, a major side effect from oral antibiotic therapy. Another area of possible advantage is the nature of cost. With Vancomycin(antibiotic for the treatment of infections caused by Gram-positive bacteria) being more expensive than phage therapy
“Pio bacteriophagum fluidum”- one of the polyvalent phage preparartions produced by the EIBMV.The preparation targets a variety of bacterial pathogens, including P.aeruginosa, E. coli, S.aureus, Streptococcus and proteus
Agriculture / food- Bacteriophage Therapy use to reduce enteric pathogen Campylobacter jejuni Industry-A broad number of food products, commodity chemicals, and biotechnology products are manufactured industrially by large-scale bacterial fermentation of various organic substrates Dentistry-In dentistry we have found that one specific germ, Strep mutans, has a direct and key role in the formation of cavities- see . Its preventive or treatment strategy to make a "phage cocktail" against tooth decay.
Limitations of phage therapy
1.Emergence of bacterial strains resistant to particular phages. The emergence of phage – resistant bacterial mutants was observed and the phenomenon was suggested to be a potential problem of phage therapy
Limitations of phage therapy
2.The development of phage–neutralizing antibodies-The production of neutralizing antibodies should not be a significant obstacle during initial or relatively short-term therapeutic treatments at least.
Limitations of phage therapy
The use of phage strains whose host range was too narrow, meant many investigators were led to false negative results after they used „off the shelf‟ phage preparations. The phage preparations used were often crude bacterial debris; filter sterilised, still containing large loads of endo- and exotoxins. These were administered orally, intravenously, intramuscularly, intraparenterally and intrathecally. Not surprisingly with the patients often in a weakened state, the
„therapy‟ often caused more harm than doing nothing.
Limitations of phage therapy
Another problem in early phage experiments was the failure to distinguish between lytic and lysogenic phages. Lysogenic phages do not always lyse the host bacterium, but integrate portions of their genome into the bacterial genome as „prophages. In order to use phage as a therapy ,the pathogen causing infection must properly diagonosed so that the appropriate phage can be used. phages are highly specific and a given phage such as the cholera phage will kill only cholera bacteria but not other. diagonosis is quite difficult.
Combating the limitations
Modernization of phage therapy 1. Sequencing of whole genome 2.Rapid and high –input, sequence –based Screening methodologies(e.g., microarrays)
We are able to tackle the increase in resistance in many common bacterial pathogens.
Methicillin resistance s.aureus Vancomycin resistant enterococci.
Genomic is providing a new strategy by revealing new molecular targets and peptides that are giving rise to novel antimicrobial drug.