BEE VENOM TREATMENT FOR MICROBES
What Are Microbes?
They are exceptionally small living things, which can only be seen through a powerful microscope – as it is impossible to see them with a naked eye. Microbes are present in air, soil, water and our bodies. There are millions of these tiny particles present on the inside and outside of our bodies.
A few microbes are harmful, while most of them are important for our general health. Explaining it further, we can say that good and a few bad microbes live together in our body and while killing the harmful ones the large population of healthy microbes also suffers.
What Are The Types Of Microbes?
Microbes have different life forms, as they dramatically have diverse characteristics and appearance.
The common types of microbes are:
What Do Harmful Microbes Do?
The percentage of harmful microbes is quite low and it is believed that only 1% of the harmful bacteria exist, which can invade our system and make us ill. Infectious diseases caused due to microbes are measles and flu, while they may contribute towards a non-infectious disease also like: coronary heart disease or some form of cancer. Microbes that can result in any type of the disease are known as pathogens.
How Do The Microbes Act?
The pathogen is a microorganism that carries the potential to cause any sort of disease. An infection is caused due to the rapid onset and proliferation of pathogenic microbes in any individual. A disease becomes prominent when the responsible infection, results in any sort of damage to the vital system or functions. An infection doesn't have to be always going to result in disease.
How Do The Microbes Affect Us?
Microbes enter our bodies to cause an infection. They can enter our system through the following sites:
Microbes after entering the body reach the target site and get attached to it so that the infection can begin. After that, they start to multiply in a very fast manner and absorb nutrients from the host (our body). It is interesting to tell here that, they avoid and survive attacks carried out by the immune system of a host.
How Are The Harmful Microbes Treated?
A harmful microbe can lend a lot of damage if it isn’t treated on time. The reason is that they multiply very quickly and can infect a large area within a short period. That is why strong medicines, like antibiotics, are prescribed to people suffering from different types of harmful microbes.
But each medicine has its side effects and one of them is that; along with the harmful microbes, they also damage the good or essential bacteria. That can affect a body's natural immune systems' strength to fight against any external harmful microbes.
For that reason, it is important to develop a natural therapeutic agent that can only target harmful microbes and protect useful bacteria. Honey bee venom is one of the best therapeutic agents which, if used in proper dose and cycle, can effectively kill the harmful microbes.
Bee Venom (Apis Mellifera) an Effective Potential Alternative to Gentamicin for Specific Bacteria Strains
Abstract
Melittin is one of the most important components of bee venom and is certainly more active against the gram-positive bacteria than the gram-negative bacteria. Various clinical studies have established that bee venom has multiple effects like anti-inflammatory, anti-viral and anti-bacterial on various types of cells. Furthermore, wasp venom also has anti-bacterial properties. The main purpose of this study was to determine the antibacterial action of bee venom against gram-positive bacteria and gram-negative bacterial strains.
For this clinical investigation, the antibacterial mechanism of bee venom was analyzed against 6-gram positive bacteria and gram-negative bacteria, which included: Burkholderia pseudomallei and Burkholderia mallei, pseudomonas aeruginosa, Escherichia coli, salmonella Typhimurium and staphylococcus aureus. 3 concentrations of crude bee venom and antibiotic disks (gentamicin), as positive controls were assessed through the disc diffusion method.
It was found that bee venom generated a noticeable antibacterial effect against Salmonella Typhimurium, Staphylococcus aureus and escherichia coli in all 3 concentrations tested. It was also found that bee venom had no positive effect on the remaining types of bacteria for any of the 3 doses tested.
Results obtained indicate that bee venom prevents the growth and survival of harmful bacterial strains. Bee venom can be used as a supplementary therapeutic anti-microbial agent against different pathogenic bacteria. However, bee venom lacked essential proteins that are important for demonstrating anti-bacterial activity for a few strains. It can also be declared that bee venom initiates a proper molecular mechanism that leads to an anti-bacterial effect on different strains of harmful bacteria. Further studies are required to understand the mechanism responsible for generating this anti-bacterial effect.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5043086/
Antimicrobial Activity of Bee Venom and Melittin against Borrelia Burgdorferi
Introduction
The percentage of infectious diseases has gone quite high, during the past few years. But the main problem is that there is no proper effective treatment, even in the presence of antibiotics, that can control these infections. Lyme borreliosis, also known as Lyme disease has affected different parts of Africa, Europe, and the USA. Lyme disease is caused due to harmful bacteria – Borrelia burgdorferi that is usually transmitted through deer ticks. Antibiotics like Ceftriaxone, Amoxicillin, and Doxycycline are usually prescribed to eliminate borrelia ssp, but they are not that effective and have numerous side effects.
Considering the limiting effect of different antibiotics on borrelial morphologies, this clinical study was conducted to observe the anti-microbial activity of bee venom and melittin against Borrelia burgdorferi. Bee venom is obtained from Apis mellifera and has been used for a long time due to its positive anti-microbial effects. In this clinical report, different forms of Borrelia burgdorferi were tested against bee venom and melittin and the effects were compared with the treatment of an anti-biotic.
To evaluate the anti-microbial activity of bee venom and melittin, an SYBR Green I/PI assay along with a total live cell counting was used for stationary phase and log phase persisters. Attached biofilms were examined through LIVE/DEAD and crystal violet staining techniques. Atomic force and fluorescent microscopy methods were used to have a better picture of the anti-microbial effects of melittin and bee venom on borrelia.
Method
Borrelia burgdorferi strain was acquired from American Type Culture Collection. Strained bacteria was maintained at very low passage isolates in the Barbour Stoner Kelly H media supplemented with a 6% rabbit serum having no antibiotics in a sterile glass and incubated at 33 degrees centigrade having 5% CO2.
Apis mellifera was prepared using a sterile 1x phosphate buffer saline, having a pH value of 7.4 for in vitro testing. Melittin extracted from BV was acquired from Sigma, USA and then prepared for testing according to the instructions of the manufacturer. Antibiotics used were Daptomycin, Cefoperazone, and Doxycycline, which were also obtained from Sigma and managed according to the instructions of the manufacturer. Furthermore, anti-microbial agents were subjected to sterilization with a 0.1µm filter unit, aliquoted and then stored at minus 20 degrees centigrade.
The effectiveness of antimicrobial treatment was assessed on a stationary and logarithm phase of b. Burgdorfer spirochetes through direct counting method and SYBR Green I/PI assay. The experiment was prepared with a 1:75 attenuation of anti-microbial cured stationary culture that was placed in anti-microbial agent-free media and then incubated for one week while maintaining the standard culture conditions. After incubation, viability was examined using direct counting method and SYBR Green I/PI assay.
Microbial agents were examined for auto fluoresce, as there were issues reported in the past in SYBR Green I/PI assay related to the detection of autofluorescence of different agents. Microbial were in placed in a 96 well plate and then tested in 100µL of a BSK-H media through SYBR Green I/PI assay. The autofluorescence value was then subtracted from the experimental values.
The efficiency of the anti-microbial agents on the attached biofilms was computed after measuring the biomass through crystal violent staining after and before anti-microbial treatment. Centrifugation was conducted at 12,000xg at room temperature for 5 minutes. Once this treatment was over, culture media was removed and the attached biofilms were treated with 500µL of 1x PBS and after that, they were collected.
Borrelia biofilm structure was further visualized after anti-microbial treatment through atomic force microscopy. The BSK-H media was detached before analyzing the biofilms. Scans were done through contact mode AFM imaging performed with Nanosurf Easyscan 2 AFM using SHOCONG probes. Later, the images were processed with the help of Gwyddion software.
The quantitative results were examined by keeping in view the median value of different readings obtained from the anti-microbial screen and also a t-test was performed and graphed with the help of Microsoft Excel software. Each experiment was independently performed for 4 times and each experiment had three samples.
Result
According to the findings made through different clinical techniques, it was established that melittin and whole bee venom have a significant effect on all of the morphological forms of Borrelia burgdorferi, as they inhibit the recovery of persisters and spirochetal cells, as it was evident by the recovery cultures examined in the antimicrobial free media. Melittin and whole bee venom also helped in reducing the viability and number of attached biofilms, which are an extremely anti-biotic resistant genre of b. burgdorferi. MIC value in concentration for melittin was in total agreement with the clinical studies conducted previously that examined effect of melittin on b. spirochetes and on other gram-negative microorganisms.
While a comparison of the effects of melittin and bee venom on borrelia demonstrated a few differences. Like the ultrastructure analyses through atomic force microscopy depicted that bee venom was more effective on the size and morphology of biofilms than its sustainability. It suggests that biofilms' response is somewhat complicated and requires a detailed study.
The difference in the level of effectiveness of melittin and bee venom on b. Burgdorfer gives a hint that there are other ingredients present in the whole bee venom, which affect the borrelia biofilms. The same findings were observed in a recent clinical study that tested stevia extracts on b. burgdorferi. It was found that a whole stevia leaf extract was effective, while its component stevioside did not deliver the same effect.
Adding further, the antibiotics that were used in this clinical study demonstrated no or little effect on the attached biofilm types of B. burgdorferi. Identical findings were found in previous clinical studies on the staphylococcus aureus and pseudomonas aeruginous, which founded that different antibiotics could not help eliminate biofilm form in different cases and might increase its size. In this clinical study, nearly the same result was observed for the antibiotic doxycycline that leads to a definite increase in the mass of attached borrelia biofilm mass.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745474/
Conclusion
Both of the clinical studies that have been discussed above indicate that bee venom or its components can help in controlling and eliminating harmful microbes. Bee venom is even effective on those microbes that are highly resistant to different antibiotics. These studies provide a platform for medical researchers to use bee venom for treating different types of microbes. Though, further studies are required that have a large sample base and include different components of bee venom and bee venom as a whole and different antibiotics. The antibiotic, if used, must be relevant to the type of microbe that is being studied. It will eventually make it easier for the health practitioners to recommend the patients, an exact dose and frequency of bee venom that can easily get rid of the harmful microbes.
They are exceptionally small living things, which can only be seen through a powerful microscope – as it is impossible to see them with a naked eye. Microbes are present in air, soil, water and our bodies. There are millions of these tiny particles present on the inside and outside of our bodies.
A few microbes are harmful, while most of them are important for our general health. Explaining it further, we can say that good and a few bad microbes live together in our body and while killing the harmful ones the large population of healthy microbes also suffers.
What Are The Types Of Microbes?
Microbes have different life forms, as they dramatically have diverse characteristics and appearance.
The common types of microbes are:
- Viruses
- Protists
- Fungi
- Archaea
- Bacteria
What Do Harmful Microbes Do?
The percentage of harmful microbes is quite low and it is believed that only 1% of the harmful bacteria exist, which can invade our system and make us ill. Infectious diseases caused due to microbes are measles and flu, while they may contribute towards a non-infectious disease also like: coronary heart disease or some form of cancer. Microbes that can result in any type of the disease are known as pathogens.
How Do The Microbes Act?
The pathogen is a microorganism that carries the potential to cause any sort of disease. An infection is caused due to the rapid onset and proliferation of pathogenic microbes in any individual. A disease becomes prominent when the responsible infection, results in any sort of damage to the vital system or functions. An infection doesn't have to be always going to result in disease.
How Do The Microbes Affect Us?
Microbes enter our bodies to cause an infection. They can enter our system through the following sites:
- Through any sort of breaks on the surface of the skin (for example Clostridium tetani that result in tetanus)
- Through urogenital tract (for example Escherichia coli that results in cystitis)
- Through gastrointestinal tract (oral cavity of the mouth) (for example Vibrio cholera that results in cholera)
- Through respiratory tract (nose or mouth) (for an example influenza virus that results in flu)
Microbes after entering the body reach the target site and get attached to it so that the infection can begin. After that, they start to multiply in a very fast manner and absorb nutrients from the host (our body). It is interesting to tell here that, they avoid and survive attacks carried out by the immune system of a host.
How Are The Harmful Microbes Treated?
A harmful microbe can lend a lot of damage if it isn’t treated on time. The reason is that they multiply very quickly and can infect a large area within a short period. That is why strong medicines, like antibiotics, are prescribed to people suffering from different types of harmful microbes.
But each medicine has its side effects and one of them is that; along with the harmful microbes, they also damage the good or essential bacteria. That can affect a body's natural immune systems' strength to fight against any external harmful microbes.
For that reason, it is important to develop a natural therapeutic agent that can only target harmful microbes and protect useful bacteria. Honey bee venom is one of the best therapeutic agents which, if used in proper dose and cycle, can effectively kill the harmful microbes.
Bee Venom (Apis Mellifera) an Effective Potential Alternative to Gentamicin for Specific Bacteria Strains
Abstract
Melittin is one of the most important components of bee venom and is certainly more active against the gram-positive bacteria than the gram-negative bacteria. Various clinical studies have established that bee venom has multiple effects like anti-inflammatory, anti-viral and anti-bacterial on various types of cells. Furthermore, wasp venom also has anti-bacterial properties. The main purpose of this study was to determine the antibacterial action of bee venom against gram-positive bacteria and gram-negative bacterial strains.
For this clinical investigation, the antibacterial mechanism of bee venom was analyzed against 6-gram positive bacteria and gram-negative bacteria, which included: Burkholderia pseudomallei and Burkholderia mallei, pseudomonas aeruginosa, Escherichia coli, salmonella Typhimurium and staphylococcus aureus. 3 concentrations of crude bee venom and antibiotic disks (gentamicin), as positive controls were assessed through the disc diffusion method.
It was found that bee venom generated a noticeable antibacterial effect against Salmonella Typhimurium, Staphylococcus aureus and escherichia coli in all 3 concentrations tested. It was also found that bee venom had no positive effect on the remaining types of bacteria for any of the 3 doses tested.
Results obtained indicate that bee venom prevents the growth and survival of harmful bacterial strains. Bee venom can be used as a supplementary therapeutic anti-microbial agent against different pathogenic bacteria. However, bee venom lacked essential proteins that are important for demonstrating anti-bacterial activity for a few strains. It can also be declared that bee venom initiates a proper molecular mechanism that leads to an anti-bacterial effect on different strains of harmful bacteria. Further studies are required to understand the mechanism responsible for generating this anti-bacterial effect.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5043086/
Antimicrobial Activity of Bee Venom and Melittin against Borrelia Burgdorferi
Introduction
The percentage of infectious diseases has gone quite high, during the past few years. But the main problem is that there is no proper effective treatment, even in the presence of antibiotics, that can control these infections. Lyme borreliosis, also known as Lyme disease has affected different parts of Africa, Europe, and the USA. Lyme disease is caused due to harmful bacteria – Borrelia burgdorferi that is usually transmitted through deer ticks. Antibiotics like Ceftriaxone, Amoxicillin, and Doxycycline are usually prescribed to eliminate borrelia ssp, but they are not that effective and have numerous side effects.
Considering the limiting effect of different antibiotics on borrelial morphologies, this clinical study was conducted to observe the anti-microbial activity of bee venom and melittin against Borrelia burgdorferi. Bee venom is obtained from Apis mellifera and has been used for a long time due to its positive anti-microbial effects. In this clinical report, different forms of Borrelia burgdorferi were tested against bee venom and melittin and the effects were compared with the treatment of an anti-biotic.
To evaluate the anti-microbial activity of bee venom and melittin, an SYBR Green I/PI assay along with a total live cell counting was used for stationary phase and log phase persisters. Attached biofilms were examined through LIVE/DEAD and crystal violet staining techniques. Atomic force and fluorescent microscopy methods were used to have a better picture of the anti-microbial effects of melittin and bee venom on borrelia.
Method
Borrelia burgdorferi strain was acquired from American Type Culture Collection. Strained bacteria was maintained at very low passage isolates in the Barbour Stoner Kelly H media supplemented with a 6% rabbit serum having no antibiotics in a sterile glass and incubated at 33 degrees centigrade having 5% CO2.
Apis mellifera was prepared using a sterile 1x phosphate buffer saline, having a pH value of 7.4 for in vitro testing. Melittin extracted from BV was acquired from Sigma, USA and then prepared for testing according to the instructions of the manufacturer. Antibiotics used were Daptomycin, Cefoperazone, and Doxycycline, which were also obtained from Sigma and managed according to the instructions of the manufacturer. Furthermore, anti-microbial agents were subjected to sterilization with a 0.1µm filter unit, aliquoted and then stored at minus 20 degrees centigrade.
The effectiveness of antimicrobial treatment was assessed on a stationary and logarithm phase of b. Burgdorfer spirochetes through direct counting method and SYBR Green I/PI assay. The experiment was prepared with a 1:75 attenuation of anti-microbial cured stationary culture that was placed in anti-microbial agent-free media and then incubated for one week while maintaining the standard culture conditions. After incubation, viability was examined using direct counting method and SYBR Green I/PI assay.
Microbial agents were examined for auto fluoresce, as there were issues reported in the past in SYBR Green I/PI assay related to the detection of autofluorescence of different agents. Microbial were in placed in a 96 well plate and then tested in 100µL of a BSK-H media through SYBR Green I/PI assay. The autofluorescence value was then subtracted from the experimental values.
The efficiency of the anti-microbial agents on the attached biofilms was computed after measuring the biomass through crystal violent staining after and before anti-microbial treatment. Centrifugation was conducted at 12,000xg at room temperature for 5 minutes. Once this treatment was over, culture media was removed and the attached biofilms were treated with 500µL of 1x PBS and after that, they were collected.
Borrelia biofilm structure was further visualized after anti-microbial treatment through atomic force microscopy. The BSK-H media was detached before analyzing the biofilms. Scans were done through contact mode AFM imaging performed with Nanosurf Easyscan 2 AFM using SHOCONG probes. Later, the images were processed with the help of Gwyddion software.
The quantitative results were examined by keeping in view the median value of different readings obtained from the anti-microbial screen and also a t-test was performed and graphed with the help of Microsoft Excel software. Each experiment was independently performed for 4 times and each experiment had three samples.
Result
According to the findings made through different clinical techniques, it was established that melittin and whole bee venom have a significant effect on all of the morphological forms of Borrelia burgdorferi, as they inhibit the recovery of persisters and spirochetal cells, as it was evident by the recovery cultures examined in the antimicrobial free media. Melittin and whole bee venom also helped in reducing the viability and number of attached biofilms, which are an extremely anti-biotic resistant genre of b. burgdorferi. MIC value in concentration for melittin was in total agreement with the clinical studies conducted previously that examined effect of melittin on b. spirochetes and on other gram-negative microorganisms.
While a comparison of the effects of melittin and bee venom on borrelia demonstrated a few differences. Like the ultrastructure analyses through atomic force microscopy depicted that bee venom was more effective on the size and morphology of biofilms than its sustainability. It suggests that biofilms' response is somewhat complicated and requires a detailed study.
The difference in the level of effectiveness of melittin and bee venom on b. Burgdorfer gives a hint that there are other ingredients present in the whole bee venom, which affect the borrelia biofilms. The same findings were observed in a recent clinical study that tested stevia extracts on b. burgdorferi. It was found that a whole stevia leaf extract was effective, while its component stevioside did not deliver the same effect.
Adding further, the antibiotics that were used in this clinical study demonstrated no or little effect on the attached biofilm types of B. burgdorferi. Identical findings were found in previous clinical studies on the staphylococcus aureus and pseudomonas aeruginous, which founded that different antibiotics could not help eliminate biofilm form in different cases and might increase its size. In this clinical study, nearly the same result was observed for the antibiotic doxycycline that leads to a definite increase in the mass of attached borrelia biofilm mass.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745474/
Conclusion
Both of the clinical studies that have been discussed above indicate that bee venom or its components can help in controlling and eliminating harmful microbes. Bee venom is even effective on those microbes that are highly resistant to different antibiotics. These studies provide a platform for medical researchers to use bee venom for treating different types of microbes. Though, further studies are required that have a large sample base and include different components of bee venom and bee venom as a whole and different antibiotics. The antibiotic, if used, must be relevant to the type of microbe that is being studied. It will eventually make it easier for the health practitioners to recommend the patients, an exact dose and frequency of bee venom that can easily get rid of the harmful microbes.
