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HEALTH Colloidal silver was ‘erased from textbooks’ because cured diseases from tuberculosis to syphilis
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  1. #41
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    Quote Originally Posted by NC Susan View Post

    In 1999, after a re-emergence of silver, the F.D.A. completely banned it in any form from being sold in over-the-counter health products, despite the fact that silver has safely been used as a medicine for millennia.

    The devastation of the polio epidemic was largely caused by how the Food and Drug Administration suppressed silver to promote its new antibiotic and vaccine industries. By removing silver from the lists of approved medicines, it effectively removed the only treatment that reliably kills polio, which in turn unleashed the full epidemic of polio. The F.D.A. later claimed a victory over polio in boasting that the new generation of vaccines had saved us.

    The agency cunningly waited until the epidemic was in its natural decline to release the vaccine, in order to ensure that people saw a connection between the vaccine’s release date and the disappearance of the disease. The public, and quite a few doctors were distracted away from the fact that silver medications were a safe treatment, which effectively kills polio quickly, as well as virtually every other virus known.

    Prior to this entire smoke-and-mirrors routine, silver was recognized for doing what the establishment now claims is impossible. Had silver medicine not been stripped from the market, the polio epidemic would have never occurred.
    As a very healthy "alternative medicine" family, for lack of a better term, we have learned that if the medical community presents a united front against any substance or treatment in particular, at the very least it is worth looking into, colloidal silver being one of those.

    To a large degree (with exceptions here and there), today's doctors are little more than a delivery system for Big Pharm products, many of which do more harm than good. Where would Big Pharm be without the doctors?

    There is a reason that organized medicine is opposed to so many proven, affordable substances and treatments, and it is most definitely not because they are concerned about your health.
    This might hurt a little.

  2. #42
    This company makes good silver impregnated dressings. Argentum Medical Llc

  3. #43
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    I'd like to see a test done where there are 5 - 6 petrie dishes where different types of "common" bacteria have been grown and then a standard solution of CS added to the dishes.

    1. Were any of the bacteria killed?
    2. Which ones?
    3. How many of the bacteria were killed? (all? some?)

    This is the sort of test that I haven't been able to find, and I do have access to medical databases in the deep web.

    Barring that, I have seen homemade CS work on eye infections in dogs and cats.

    On the other hand, I've never seen it work on poison ivy or sore throats. I have seen elderberry stop a sore throat, that was likely a viral infection, in its tracks.

    So, I don't know. I'm not made of money, and if I can try a home remedy first, I generally will. On the other hand, there are times when it pays to get to a doctor quickly.

    I pray that God gives me the wisdom to know which is best.

  4. #44
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    Quote Originally Posted by Daytonabill0001 View Post
    Not taking sides here, but how would you explain this statement from the article NC Susan posted?
    I owe you an explanation as you asked fairly and decently. I thank you for that.

    First, I need to quote a section from the article:

    The most popularized case of argyria is that of Paul Karason, the so-called Smurf Man. He internally consumed large doses of a homemade silver solution for years, and then began also using silver externally on his face. He too made his silver solution using salt, which resulted in silver chloride.

    Despite it being a completely different substance, he refers to his solution as “colloidal silver”, as does the media. We believe that after he noticed some slight skin discoloration, he actually increased his dosage, because he had found a way to become famous and profit from being a freak.

    He admits publicly to using excessive amounts of his silver chloride both internally and transdermally, daily for 14 years. He still continues to use it, despite his obvious saturation. He further admits that his face turned blue before the rest of his body; and yet he continued to use both silver products, despite the color change.

    Due to Karason’s self-inflicted and intentional cosmetic alteration, the F.D.A. has been using him in a public relations campaign that is intended to convince the public that colloidal silver is dangerous. We can only speculate about how much the media networks and the F.D.A. have paid him for his appearances.
    I had never run across this particular fellow before and had to research him because the story struck me as odd. In truth, per his family he was reclusive before the 'change' and others noticed it before he did, well before. He never sought fame and remained reclusive, but did agree to a couple of interviews.

    I did not find any indications that he was trotted out by the FDA, etc, but ABC News, the Today Show, and Fox News did stories on him. All indications were that he was sought out rather than trying to exploit his condition for fame and fortune.

    Note the bolded section. This is from the article dated this month, the report on Mr. Karason. Paul Karason died of a heart attack at age 62 in September of 2013. So I am unsure as to how he might still be using the products. This is just one little bit of inaccuracy I found in the article.

    You also quoted and asked about:

    However, every case that we investigated involved products that were not actually colloidal silver, and most cases were the result of pharmaceutical-industry silver products. Our exhaustive research could not find a single instance of argyria that was caused by pure colloidal silver. The pharmaceutical silver solutions are the most likely to cause it.
    First, most cases occurring since the mid-1990's that I can find, and there are a few to be certain, were the result of using home brew colloidal silver such as touted by enthusiasts who make their own using various and sundry 'generators'.

    Pharmaceutical industry silver products would be Silvadene cream, and Flamazine in the British Empire countries. Both are topical applications used for wound management, especially for burns. There have been reports of localized argyria following repeated application of them. Localized meaning surrounding the wound area and within the new skin over the wound area itself. I cannot find any cases of systemic affect.

    There are of course silver-impregnated dressings which are not considered to be pharmaceutical products per se. i.e. they do not require a prescription for use such as silver-containing salves do. Silver has long been known to work in vitro, i.e. 'on the surface,' but have never been proved to work in vivo (internally).

    There are no pharmaceutical industry-produced internal products. Those are produced ENTIRELY by the 'nutritional supplements' industry.

    BTW, it might interest some to know that just since 2010 there have been several animal studies which show that silver does accumulate in the organs, including the brain, and that it can damage the kidneys in particular.

    Now, at the risk of being accused of thread drift, I am going to say that the new search engine that was recommended here on TB2K sometime last week I believe has been a boon. Google doesn't even begin to show some of the returns Startpage does. Likewise DuckDuckGo.

    Daytona, I hope this answers your questions. If not, you know where to find me.

    RR
    Survival and Austere Medicine: An Introduction - 2nd and 3rd Editions Contributing author and editor

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  5. #45
    I use iodine for skin infections and stuff close to the skin. Tried silver and found it useless. Tried means I turned blue-blooded.

  6. #46
    Quote Originally Posted by Reasonable Rascal View Post

    Daytona, I hope this answers your questions. If not, you know where to find me.

    RR
    Fair enough Rascal, you're a good man...

    i haven't developed a habit of drinking CS because I think your immune system would develop a reliance on it to the point that it slacks off because the CS is carrying the load...

    However, one time I had a really bad time of getting rid of the creeping crud I had in my lungs, heck I couldn't take full breaths because of the pain...

    So, I bought a battery powered nebulizer and used it with the CS I made (same stuff that my DS's friend used) for 15 min on the hour and by the 2nd day, the pain left and a week later, all of the crud was gone... I had it for 3 weeks before I started nebulizing...

    I think that going to extremes or generating CS improperly would be deleterious to your health, rather it should be appreciated for what help it provides and once the crisis is over, ween yourself of off it and let your immune system take over...

    It works on toothaches for me (use it as if you were oil pulling for 15 or 20 min) and then it comes back a few months later, which is nice because it gives you time to schedule an appointment with the dentist...

    I still think that CS would and should be part of any serious medical bugout bag...
    Rom 4:8 Blessed is the man to whom the Lord will not impute sin. By the Blood of Jesus, that's me!

    Did you know that the first two chapters of Hebrews shows that the Father declared Jesus as GOD when Jesus ascended on high? :)

  7. #47
    Quote Originally Posted by China Connection View Post
    I use iodine for skin infections and stuff close to the skin. Tried silver and found it useless. Tried means I turned blue-blooded.
    Your CS was worthless because it either didn't have any colloidal silver or was way too weak. It requires at minimum, a concentration of 20 ppm to be effective against bacteria... Was it store bought?

    But for skin purposes, your style is good and provides iodine for your thyroid... Especially if the color disappears quickly, signifying a possible shortage of iodine in your body...
    Rom 4:8 Blessed is the man to whom the Lord will not impute sin. By the Blood of Jesus, that's me!

    Did you know that the first two chapters of Hebrews shows that the Father declared Jesus as GOD when Jesus ascended on high? :)

  8. #48
    BUMP because of Venezuela happenings (i.e. cholera?!??), and it conjunction with the iodine thread I am going to post.


    51 Amazing Colloidal Silver Benefits


    Silver is an ancient and natural all-purpose healing agent. It wipes out bacterial infections in a safe and clean manner, without compromising our immune system like antibiotics do.

    Silver effectively vanished from the medicinal market in the early 20th century, because it could not be patented and thereby couldn’t fetch extraordinary profits for the pharmaceutical industry. In modern lab tests (e.g. by NCBI), silver is getting its fame back as a safe, multi-purpose bacterial killer. It is known to kill superbugs (e.g. MRSA) and has been tested against 650 common and exotic infections and diseases.

    If you ever had to take antibiotics for any of the 51 conditions below (research linked), then it’s time to consider silver, whether it’s in colloidal form (made by Silver Healer) or chelated , protein-binding form for added bioavailability (made by Silver Excelsior).

    The fact is: repeated antibiotic use strengthens the bacterial strains, and radically weakens the immune system.

    51 Researched Benefits Of Silver

    1. Silver has shown the ability to wipe out common throat ailments such strep.

    2. Ten drops of silver in the ear can aid in wiping out ear infections.

    3. Quality colloidal silver soothes and wipes out eye infections (conjunctivitis, stys).

    4. Colloidal silver has been shown to treat rhinosinitus in sheep, suggesting it can clears sinus and upper respiratory infections.

    5. Silver ions have shown promise in inactivating influenza and maybe show promise for inactivating other viruses.

    6. Silver nano-particles have shown success in promoting anti-inflammatory properties.

    7. Silver-coated endotracheal tubes have shown promise in prevention of ventilator-associated pneumonia.

    8. Placing silver induced products on the urinary tract has shown promise in treating Urinary Tract Infections (UTIs)

    9. Over time, antibiotics lose their ability to combat infection. Silver has been shown to boost the immune system to fight infection.

    10. Silver nanoparticles have shown promise against hematophagous parasites.

    11. Swishing silver in the mouth has been shown to prevent plaque buildup, tooth decay, and bleeding gums.

    12. Silver promotes optimal digestion.

    13. Digested silver quickly combats food poisoning.

    14. Silver has the ability to kill mold (including black mold) and mildew on surfaces throughout the home.

    15. Applying silver products on cuts, sores, and boils promotes healing.

    16. Applying silver to rashes can aid in soothing itches.

    17. Silver has the ability to kill various fungi, including athlete’s foot and nail fungi.

    18. Silver has been successfully used to kill staph infections, including MRSA.

    19. Applying silver to burns, especially sunburns, can soothe and heal.

    20. Silver has shown success in inactivating the herpes simplex virus.

    21. Silver infused water has been used to support HIV/AIDS intervention.

    22. Silver has been shown to kill cancerous cells and has been considered as an alternative treatment to chemotherapy.

    23. Silver applied to insect bites provides relief from itches.

    24. Silver applied to infected areas can kill ringworms.

    25. Silver applied to the vaginal area can combat yeast infections.

    26. Relief from jock itch can be found through applying silver around the affected area.

    27. Silver’s antibacterial properties can provide relief from acne breakouts.

    28. Applying silver to the scalp can relieve itchy scalp and dandruff.

    29. When ingested, silver shows promise in treating laryngitis.

    30. Silver’s ability to kills single cells organisms allows it to treat eczema when applied to the affected area.

    31. Silver shows promise in treating psoriasis.

    32. Silver purifies water that has germ contaminants present.

    33. Silver is used in many airline filters to prevent water borne diseases.

    34. When applied on mayonnaise and dairy products, silver has the ability to prevent salmonella growth.

    35. Ketchup, mustard, relish, and other condiments get a prolonged life with the addition of silver.

    36. Silver’s anti-fungal, anti-bacterial, and anti-viral can help treat Lyme disease.

    37. Silver’s antiviral properties help combat the common cold.

    38. When applied to the scalp, silver can promote hair growth.

    39. Sixty-six percent of patients in a trial found that their warts regressed when silver was applied.

    40. Silver’s anti-microbial properties have shown promise in stopping asthma attacks.

    41. When ingested, silver has the ability to combat Whooping cough.

    42. Using colloidal silver regularly allowed one woman to treat her Multiple Sclerosis.

    43. Can minimize symptoms of Crohn’s disease.

    44. When ingested silver can heal a pet’s stomach ache.

    45. Silver can heal pet skin issues such as rashes and ringworm.

    46. When pets leave messes on the floor or carpet, silver can get rid of nasty odors.

    47. Applying silver to cuts helps treats both human and pet infections.

    48. Applying silver on cuts can prevent infections in pets.

    49. Silver has the ability to keep pets fungi-free.

    50. Colloidal silver can penetrate the large intestine and treat traveler’s diarrhea and cholera.

    51. Silver has the ability to inhibit both Hepatitis B & C viruses.

    Colloidal Silver Benefits



    https://www.honeycolony.com/article/...its-of-silver/
    Romans 12:14 Ask God to bless those who persecute you—yes, ask Him to bless, not to curse.

  9. #49
    I'm going to provide some more baseless anecdotal evidence here... take it for what it's worth.

    An Amish friend has suffered with severe chronic cellulitis for over 10 years. She's a heavy woman, and works long hours on her feet. We happened to be talking about her brother's horse (the one i mentioned in a previous post on this thread) and she asked, "do you think that silver stuff would help my leg?"... as she lifted her skirt and unwrapped an ace bandage to reveal a leg that was purple from the knee down, and had three full thickness ulcers the size of a quarter. It was bad... frighteningly so.

    She apparently got no help from a couple of local doctors and gave up, but I honestly don't know how she was able to walk, much less do the work she does.

    She decided to try the silver for the severe chronic cellulitis, and we mixed up some Betadine and honey for the ulcers. It was about a month later when she asked if i had more silver. I asked if it was helping, and she lifted her skirt to show me... the ulcers were nearly healed... two were pink scars and one was still scabbed, but nearly healed. The swelling in the leg was down by probably 3/4, and it had faded to a sort of angry pink.

    She's still taking the silver (5 months in).... no signs of argyria, but the cellulitis is slowly receding down the leg... she thinks it might take a year, but she's so thrilled with the improvement in pain, she's planning on continuing to use it until the leg is healthy, or until all improvement plateaus to what is obviously the "new normal".

    I honestly have to wonder if studies aren't being corrupted and positive info is being actively suppressed. I never would have believed that could possibly be true, but what passes for science snd ethics these days are nearly unrecognizable to old farts like me. And when I... bending over backwards to believe it *DOESN'T * work in the beginning, and only cautiously using it in animals who essentially won't live without a miracle... see it work time after time, treating everything from cellulitis to pneumonia to bacterial scours in calves... SOMEONE doesn't want this knowledge out.

    And if I could get a grant from somewhere yo run a double blind study on using intrauterine colloidal silver to treat postpartum uterine infections, I could settle a good part of this argument pretty quickly. Dairy farms already keep the relevant records (days to first heat, days to conception, # of services per conception, etc). All we'd have to do is get someone else to put colloidal silver in one jar and plain water in another.... use them alternately on every cow with an infection for 6 months, compare groups and then unmask which is which.

    But in the meantime... it's a very valuable part of my medical preps.

    Summerthyme

  10. #50
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    Quote Originally Posted by Reasonable Rascal View Post
    I have seen a lot of extremely ignorant remarks made on these forums over the years but you, sir, have just topped them all for the degree of juvenile-level immaturity exhibited in your statements.

    If you have been actually reading the forums over the years and not just mouthing off you'd know I do not have to offer my bona fides to you. They have been well enough established over the years, well before you ever appeared on the scene.

    Unlike others I *HAVE* done the research you speak of. I *HAVE* studied micro, biology, chemistry and the medical sciences, and I possess two active medical licenses. I have practiced medicine for over 40 years, published two medical books in concert with other highly qualified authors, and have living breathing functioning former patients who that could not be said of when I first encountered them.

    The amount of qualified research that has been done on colloidal silver in particular, as well as exposure to silver in general, just since 2010, is significant. Some of us actually know how to interpret the study results and do not rely on conspiracy theory to determine what medical approaches we will take.

    Why don't you stop hiding behind your keyboard, get out in the real world and do something with your life besides warming your chair.

    RR
    Didn't you used to drive an Ambulance ?

  11. #51
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    Quote Originally Posted by Dredge View Post
    Didn't you used to drive an Ambulance ?
    Point being? So did my parents' PCP - worked her way through pre-med as a Paramedic.

    RR
    Survival and Austere Medicine: An Introduction - 2nd and 3rd Editions Contributing author and editor

    Get your FREE copy of the 3rd edition here:NNPG Download Site

    Reddit AustereMedicine/

  12. #52
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    I had polio when I was 8 years old. I had my tonsils until I was 10 years old.
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  13. #53
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    Quote Originally Posted by Lei View Post
    I had polio when I was 8 years old. I had my tonsils until I was 10 years old.
    Relevance?
    Last edited by Profit of Doom; 03-13-2019 at 01:33 AM.
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  14. #54
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    Didn't do a thing for me with the Catfish Foot. :shrug:
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  15. #55
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    Grandma Sez:

    TIP for those that PROPERLY MAKE their own pure "colloidal" (ACTUALLY IONIC SILVER SOLUTION) AT HOME.
    YOU can REALLY "speed up" making the NEXT batch of ionic silver, by putting even a little (1/2 to 1 cup) of the previous batch of colloidal silver you made into the new distilled water( and stir) before you insert the electrodes and start making a new batch!!

    Actually, I am surprised nobody else (that I have come across) has realized that fact and mentioned it before!
    You will start seeing those "little clouds" of ions leaving the electrode MUCH MUCH sooner!
    Last edited by ainitfunny; 03-13-2019 at 03:45 AM.
    “SAVED” or “UNSAVED”, Jesus knows you personally,
    everything you are going through right now,
    and He loves you Dearly, and wants to help.

  16. #56
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    Quote Originally Posted by ainitfunny View Post
    Grandma Sez:

    TIP for those that PROPERLY MAKE their own pure "colloidal" (ACTUALLY IONIC SILVER SOLUTION) AT HOME.
    YOU can REALLY "speed up" making the NEXT batch of ionic silver, by putting even a little (1/2 to 1 cup) of the previous batch of colloidal silver you made into the new distilled water( and stir) before you insert the electrodes and start making a new batch!!

    Actually, I am surprised nobody else (that I have come across) has realized that fact and mentioned it before!
    You will start seeing those "little clouds" of ions leaving the electrode MUCH MUCH sooner!
    Very cool tip. I will try that this weekend

  17. #57
    I have found cs mentioned in old veterinary books. It was commonly used as a topical antiseptic/antibacterial treatment on wounds, eye infections, and skin ailments. It eventually fell out of favor when antibiotics became widely available.
    "This place is fantastic! It's like "Gone With The Wind" on mescaline. They walk imaginary pets here...and they're all heavily armed and drunk..."
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  18. #58
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    Meem, the BYU study is out there.

    it's actually pretty cool as for effectiveness shown.
    RULE 1:
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  19. #59
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    Quote Originally Posted by ainitfunny View Post
    Grandma Sez:

    TIP for those that PROPERLY MAKE their own pure "colloidal" (ACTUALLY IONIC SILVER SOLUTION) AT HOME.
    YOU can REALLY "speed up" making the NEXT batch of ionic silver, by putting even a little (1/2 to 1 cup) of the previous batch of colloidal silver you made into the new distilled water( and stir) before you insert the electrodes and start making a new batch!!

    Actually, I am surprised nobody else (that I have come across) has realized that fact and mentioned it before!
    You will start seeing those "little clouds" of ions leaving the electrode MUCH MUCH sooner!
    Quote Originally Posted by Warm Wisconsin View Post
    Very cool tip. I will try that this weekend
    This works because you are jump-starting the ion exchange (current flow). Distilled water is NOTORIOUSLY difficult to start a current flow in. This helps.
    RULE 1:
    THEY want you DEAD.

    "If there must be trouble, let it be in my day, that my brothers' children (and their parents) may have peace, and have NO KNOWLEDGE of what I have done."

    TACAMO!! NOW!!

  20. #60
    Quote Originally Posted by big kumara View Post
    I have found cs mentioned in old veterinary books. It was commonly used as a topical antiseptic/antibacterial treatment on wounds, eye infections, and skin ailments. It eventually fell out of favor when antibiotics became widely available.
    Antibiotics. Yeah. The thread I started on iodine yesterday, and will add to soon, discusses what they put in antibiotics (and other things) that mimics iodine.
    Each and every cell in our bodies has a receptor that uses iodine. TPTB made sure to replace iodine with three harmful mimickers that contribute to the dumbing down of society and affect the pineal gland...those 3 are fluorine, bromine, and chlorine.

    Re: CS, I have used for many, many years with no adverse effects.
    Romans 12:14 Ask God to bless those who persecute you—yes, ask Him to bless, not to curse.

  21. #61
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    Bumped for those who did not see the TIP as to how to make ionic silvermaking go faster, in post #55 above!
    “SAVED” or “UNSAVED”, Jesus knows you personally,
    everything you are going through right now,
    and He loves you Dearly, and wants to help.

  22. #62
    CS cured my youngest sons molluscum when nothing else the doctor gave him would.
    "...Awake thou that sleepest, and arise from the dead, and Christ shall give thee light. See then that ye walk circumspectly, not as fools, but as wise, Redeeming the time, because the days are evil." - Ephesians 5:14-17

  23. #63
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    RR and I fought this battle together many times as co-mods over at AWRM. CS has as much scientific evidence backing up its effectiveness as does stump water gathered under a full moon (to steal from Mark Twain). If folks want to bet their lives (or the lives of their families) on it, be my guest. Other than an article published in Lancet many years ago, which showed that it works on a few organisms in a petri dish, there is zip evidence that it works at all "in vivo". It will, however, turn folks who overuse it a lovely shade of blue, permanently.
    If at first you don't secede, try, try again!

  24. #64
    Folks, if silver was the curative treatment as claimed in this thread, it would have been so inculcated into medical practice that there is no way that Big Pharm or anybody else could have driven it out. So I must be missing something. What is it?
    "The misfortune of many is the consolation of fools" Ancient proverb

  25. #65
    Quote Originally Posted by Jonas Parker View Post
    RR and I fought this battle together many times as co-mods over at AWRM. CS has as much scientific evidence backing up its effectiveness as does stump water gathered under a full moon (to steal from Mark Twain). If folks want to bet their lives (or the lives of their families) on it, be my guest. Other than an article published in Lancet many years ago, which showed that it works on a few organisms in a petri dish, there is zip evidence that it works at all "in vivo". It will, however, turn folks who overuse it a lovely shade of blue, permanently.
    True CS does not turn folks blue. However, silver proteins can.

    Oprah is a globalist. She had the blue man on her show. He claimed CS did it. A lie.
    Romans 12:14 Ask God to bless those who persecute you—yes, ask Him to bless, not to curse.

  26. #66
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    Quote Originally Posted by Jonas Parker View Post
    RR and I fought this battle together many times as co-mods over at AWRM. CS has as much scientific evidence backing up its effectiveness as does stump water gathered under a full moon (to steal from Mark Twain). If folks want to bet their lives (or the lives of their families) on it, be my guest. Other than an article published in Lancet many years ago, which showed that it works on a few organisms in a petri dish, there is zip evidence that it works at all "in vivo". It will, however, turn folks who overuse it a lovely shade of blue, permanently.
    While you have a perfect right to espouse your opinions, they can prudently be ignored because they are based on ignorance. You simply don't know what you are talking about.

    Maybe if you actually READ a US Government Medical Research Report, (National Institute of Health) IF you have any respect for they Truth, you will change your mind. Here is one to start with:

    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4740956/

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    Journal ListJundishapur J Microbiolv.8(11); 2015 NovPMC4740956

    Logo of jjm
    Jundishapur J Microbiol. 2015 Nov; 8(11): e25867.
    Published online 2015 Nov 21. doi: 10.5812/jjm.25867
    PMCID: PMC4740956
    PMID: 26862383
    Bactericidal and Antibiotic Synergistic Effect of Nanosilver Against Methicillin-Resistant Staphylococcus aureus
    Khalid A. Ali Abdel Rahim1,2,* and Ahmed Mohamed Ali Mohamed3
    Author information Article notes Copyright and License information Disclaimer
    This article has been cited by other articles in PMC.
    Go to:
    Abstract
    Background:

    Methicillin-Resistant Staphylococcus aureus (MRSA) are bacteria responsible for several difficult-to-treat infections in humans. These strains have developed, through the process of natural selection. Infections by MRSA are more difficult to treat with standard types of antibiotics and thus more dangerous to human health.
    Objectives:

    The aim of this study was to evaluate the bactericidal and antibiotic synergistic effect of silver nanoparticles (Ag-NPs) against MRSA.
    Materials and Methods:

    Methicillin-Resistant Staphylococcus aureus strains were isolated from clinical samples and identified, and their susceptibility was tested using the MicroScan® WalkAway-96® SI System. minimum inhibitory concentration (MIC) was determined by a microdilution method. Time kill assay was performed by exposing the MRSA isolates to different concentrations of Ag-NPs and monitoring bacterial growth, by measuring optical density at 600 nm. Tissue culture plate was used for determination of the efficacy of Ag-NPs and their combination with antibiotics in the elimination of formed biofilm.
    Results:

    The MIC value of Ag-NPs against MRSA was 100 μg/mL. Methicillin-Resistant Staphylococcus aureus cells were treated with 50, 100 and 200 µg/mL of Ag-NPs and inhibited bacterial growth so that after four hours, almost all treated MRSA cells were dead. All combinations showed effectiveness against MRSA. It was observed that MRSA did not show inhibition zones with ampicillin alone.
    Conclusions:

    Silver Nanoparticles have high therapeutic activity against MRSA, thus can be suggested as an alternative or adjuvant with antibiotics for MRSA treatment. Further studies are required to understand the synergistic effect of Ag-NPs combinations and to assess the safety and efficacy of new antibiotic-Ag-NPs combinations.
    Keywords: Ag-NPs, Nanosilver, Antibiotics, Methicillin-Resistant Staphylococcus aureus
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    1. Background

    Methicillin-Resistant Staphylococcus aureus (MRSA) is a pathogen that has received great medical interest, as it has not been eliminated from healthcare institutions or the environment (1). A bacteremia caused by S. aureus leads to between 25% and 63% mortality; the medical importance of MRSA is due to the high rate of mortality and morbidity associated with its infections and it is considered the main cause of nosocomial infections worldwide (2). The world health organization has reported an MRSA prevalence of about 70% to 80% of all the S. aureus isolates in some Asian countries (3). A total of 92 (26.3%) S. aureus isolates were obtained, of which 33 (35.9%) were MRSA and 27 (29.3%) were Multi-Drug Resistant Staphylococcus aureus (MDRSA) in Iran (4). Currently there is a critical need to find alternative antimicrobials, especially those that could be used for treatment of MRSA infections (5). Amongst several antimicrobial compounds that have been investigated, Ag-NPs showed promising antimicrobial properties, which could be used for treatment MRSA and other drug-resistant infections.

    Silver Nanoparticles (Ag-NPs) (less than 100 nm) have shown novel antimicrobial activity to a wide range of microorganisms due to their high surface area to volume ratio and their unique chemical and physical properties when compared to the properties of their bulk form (6). These Ag-NPs could be applied safely in therapy when effective concentrations against many types of organisms have been determined. Recently, we demonstrated that Ag-NPs show antifungal activities, including apoptotic cell death. In particular, we showed that hydroxyl radicals play an important role in apoptosis (7). In addition, many researches have investigated the synergistic effect of Ag-NPs when combined with other compounds: a combination of amoxicillin and Ag-NPs showed better antibacterial properties against Escherichia coli than when they were applied alone (8). The production of biofilm is related to antibiotic resistance problems (9). Bacteria strongly adhere to surfaces using a biofilm matrix, 3D, gel-like, highly hydrated and locally charged environment. Adhesion of these bacteria to other organs may contribute to the pathogenesis of infection (10). Biofilms are often associated with dental plaques (11, 12), endocarditis (13), lung infection (14) and infection through medical devices (15, 16).
    Go to:
    2. Objectives

    In the present study, we investigated the antimicrobial and anti-biofilm activities of Ag-NPs alone and in combination with conventional antibiotics against pathogenic bacteria.
    Go to:
    3. Materials and Methods
    3.1. Bacterial Isolates

    Eighty S. aureus isolates were obtained from clinical specimens at the microbiology laboratory of the Department of Clinical Analysis at the Gynecology and Children Hospital, Hafr Elbatin, KSA. Mid-stream urine, suction tip, pus and blood specimens were collected aseptically for bacteriological examination. Handling, transporting and storing of collected samples were made at refrigeration temperature. All samples were inoculated on blood agar, incubated at 37°C overnight, and colonies were processed.
    3.2. Antimbiogram and Methicillin-Resistant Staphylococcus aureus Identification

    The MicroScan® WalkAway-96® SI System (Siemens Healthcare Diagnostics Inc, USA) was used for laboratory identification, while antimicrobial susceptibility testing and MRSA detection was performed using Neg/BP/Combo 30-B1017-306E® combination panels (Siemens Healthcare Diagnostics Inc, USA). All procedures were performed according to the manufacturer’s instructions. The well-diffusion method was used to evaluate synergistic effects of Ag-NPs with antibiotics.
    3.3. Antimicrobial Activity of Silver Nanoparticles

    Silver Nanoparticles were obtained from Sigma (St. Louis, MO, USA). Minimum inhibitory concentration (MIC) was defined as the lowest concentration of Ag-NPs preparation that prevented bacterial growth. The MIC was determined using the micro-dilution method; using LB broth (Sigma, St. Louis, MO, USA) MRSA concentration was adjusted to the McFarland standard. The MRSA isolates were exposed to serial twofold dilutions of Ag-NPs, and MRSA viability was measured after 24 hours of incubation. The MRSA cell viability was measured using a colony-forming capacity assay on nutrient agar (17). All the assays were done with a negative and a positive control.
    3.4. Time- Kill Assay

    To examine the growth curves of MRSA, isolates were incubated in Mueller-Hinton broth (Sigma, St. Louis, MO, USA) with different concentrations of Ag-NPs (0, 50, 100, and 200 μg/ml), and the MRSA cells concentration was adjusted to McFarland standard and cultures were incubated in a shaking incubator at 37°C for 24 hours. Growth curves of MRSA cultures were constructed by measuring of the optical density (OD) at 600 nm at one-hour intervals for seven hours.
    3.5. Synergistic Effect of Silver Nanoparticles With Antibiotics

    The well diffusion method was used to evaluate the synergistic of Ag-NPs with antibiotics, based on the Clinical and Laboratory Science Institute (CLSI) standards (18). Briefly, Mueller-Hinton agar (Sigma, St. Louis, MO, USA) plates were inoculated with the turbidity-adjusted MRSA suspension and an antibiotic (HiMedia Chemicals Pvt. Ltd., Mumbai, India), and were incubated at 37°C for 24 hours. Next, the inhibition zone (ZOI) diameter of Ag-NPs, antibiotic and Ag-NPs + antibiotic combination were measured, by subtracting the well diameter from the total inhibition zone diameter. Synergistic effect was calculated by the following equation:
    Synergisticeffect=(B−A)(A)×100
    (1)

    Where A is ZOI for the antibiotic and B is ZOI for the antibiotic + Ag-NPs (19).
    3.6. Anti-Biofilm Activity of Nanosilver by the Tissue Culture Plate (TCP) Method

    To investigate the anti-biofilm activity of Ag-NPs alone and in combination with antibiotics, a TCP assay was performed (20). Individual wells of sterile, polystyrene, 96-well, flat-bottomed TCPs were filled with 190 µL of the bacterial suspension at 105 CFU. After 18 hours of overnight culture, Ag-NPs alone or in combination with antibiotic was added with the final concentration being the MIC. The Ag-NPs were replaced by deionized water in the control well. The TCPs were incubated for eight hours at 37°C, and the wells were then washed four times with 0.2 ml of PBS, heat-fixed at 60°C for one hour and stained with 0.4% crystal violet for 15 minutes. The stain was rinsed off, resolubilized with DMSO (Sigma, St. Louis, MO, USA) and absorbance was measured at 570 nm (Spectramax M2e, Molecular Devices, CA, USA). The control was considered to represent 100% of biofilm formation. The percentage of biofilm inhibition was calculated using the following equation (21):
    Biofilminhibition(%)=1−(Absorbanceofcellstreatedwi thsilvernanoparticles)Absorbanceofnon−treatedcontr ol×100
    (2)
    3.7. Statistical Analysis

    All experiments were performed in triplicates, and means were determined. Results were expressed as percentages.
    Go to:
    4. Results
    4.1. Bacterial Strains

    Of the eighty S. aureus isolates, 30 were MRSA, Methicillin-Resistant Staphylococcus aureus.
    4.2. Antimicrobial Activity of Silver Nanoparticles

    The MRSA isolates were exposed to twofold Ag-NPs serial dilutions for 24 hours. Silver Nanoparticles affected MRSA cellular viability in a dose-dependent manner. Cell viability assay was used to determine the bactericidal effect of different concentrations of silver nanoparticles on MRSA. Bacterial cell viability was measured using a colony-forming capacity assay in nutrient agar. The MRSA isolates were inhibited at concentrations over 100 µg/mL at 105 CFU where no visible bacterial growth was observed in the agar plates Figure 1.
    An external file that holds a picture, illustration, etc. Object name is jjm-08-11-25867-g001.jpg
    Open in a separate window
    Figure 1.
    Colony-Forming Capacity Assay

    1) Positive control, 2) growth at concentration of 12.5 µg/mL of Ag-NPs,3) growth at 25 µg/mL, 4) growth at concentration of 50 µg/ml, 5) growth at 100 µg/mL, 6) negative control. The MRSA isolates were inhibited at the concentration of 100 µg/mL at 105 CFU where no visible bacterial growth was observed in the agar plates.
    4.3. Time-Kill Assay

    Time-kill assay showed that Ag-NPs could inhibit the growth of MRSA. The growth curves of MRSA treated with 50, 100 and 200 µg/mL Ag-NPs were affected in a concentration-dependent manner and after four hours, almost all treated MRSA were dead. There was a small difference between growth of MRSA treated with 50 µg/mL Ag-NPs and the control group Figure 2.
    An external file that holds a picture, illustration, etc. Object name is jjm-08-11-25867-i001.jpg
    Figure 2.
    Methicillin-Resistant Staphylococcus aureus Growth Curve With Different Concentrations of Silver Nanoparticles
    4.4. Synergistic Effect of Nanosilver With Antibiotics

    The synergistic effects of Ag-NPs were investigated with six conventional antibiotics against MRSA, using the well diffusion method and the effects were evaluated by determination of synergism percentage. The results of the combination assay are presented in Figure 3. All of the combinations showed effectiveness against the MRSA. It was observed that MRSA did not show an inhibition zone with ampicillin alone and the highest synergistic effect was observed in the presence of clindamycine Figure 4.
    An external file that holds a picture, illustration, etc. Object name is jjm-08-11-25867-i002.jpg
    Figure 3.
    Average Inhibition Zone (mm) In the Presence of an Antibiotic, and Antibiotic Plus Silver Nanoparticles
    An external file that holds a picture, illustration, etc. Object name is jjm-08-11-25867-g002.jpg
    Open in a separate window
    Figure 4.
    Synergistic Effect of Nano-Silver (Ag-Nps) With Clindamycin
    4.5. Determination of Biofilm Formation by the Tissue Culture Plate (TCP) Method

    To examine the anti-biofilm activity of Ag-NPs and their combination with antibiotics, MRSA were grown to form biofilms and then exposed to Ag-NPs alone or in combination with the antibiotics (Figure 5). The results showed that Ag-NPs had an inhibitory activity on biofilm formation greater than 55%. Combinations of Ag-NPs and antibiotics, showed a greater inhibitory activity than Ag-NPs alone, yet ampicillin did not show any increase in activity and anti-biofilm activity was due to Ag-NPs.
    An external file that holds a picture, illustration, etc. Object name is jjm-08-11-25867-i003.jpg
    Figure 5.
    Inhibition of Methicillin-Resistant Staphylococcus aureus Biofilm Formation Determined Using the Tissue Culture Plate Method
    Go to:
    5. Discussion

    The increased resistance to antibiotics is alarming as treating infections of resistance strains could be problematic. Methicillin-Resistant Staphylococcus aureus are important pathogens with increasing rates of multi-antibiotic resistance due to both intrinsic and acquired mechanisms. This study aimed to examine the antibacterial activity of Ag-NPs against MRSA, using different in vitro assays. To achieve this goal, we challenged thirty clinical isolates classified as MRSA with different concentrations of Ag-NPs and described the effect on MRSA viability, growth rate and biofilm formation. Also we studied the synergistic effect of Ag-NPs with antibiotics to find new formulas of more effective antibiotics to overcome the antibiotic resistance problem.

    The results showed that the MRSA isolates were fully susceptible to the antibacterial effect of Ag-NPs over the MIC concentration. This proves the results obtained by other authors, which showed colloidal silver is a powerful antibiotic against a wide range of microorganisms at a very low concentration without having any harmful effect on body tissues (22, 23). Small Ag-NPs with large surface areas provide an effective antimicrobial agent even at very low concentrations. Silver Nanoparticles (diameter of 5 - 100 nm) enhances the antibacterial activity of various antibiotics (24, 25). Our results showed a 100% influence by nanosilver at the MIC of 100 µg/mL. These findings indicate that the antibacterial activity of 50 μg/mL of Ag-NPs could slightly inhibit bacterial growth yet not enough to outpace the speed of reproduction of the bacterial cells. Numerous researches have been performed studying the effect of Ag-NPs on pathogenic microorganisms. Our results showed a synergistic effect between nanosilver and antibiotics (Figures 3 and ​and44).

    These results are somewhat similar to the results obtained by Fayaz et al. (26) and Rai et al. (27) on Pseudomonas aeruginosa. Interestingly, amongst the selected antibiotics, clindamycin in combination with Ag-NPs showed the highest activity against MRSA (42%), followed by vancomycin (40%) and erythromycin (31%). The synergistic effect of Ag-NPs and clindamycin was greater when compared to other antibiotics, which may be due to both, the inhibiting protein synthesis action of the clindamycin and the DNA binding action of the Ag-NPs; similar results were obtained by Shahverdi et al. (28). The probable mechanism involved in the synergistic effect of antibiotics with Ag-NPs could be the formation of complexes between the antibiotics and Ag-NPs; though the bonding of the antibiotic’s active and functional groups like hydroxyl and amino groups to the large surface area of Ag-NPs by chelating (26, 29). Silver Nanoparticles antibacterial mechanism works by inhibiting oxygen metabolism, which finally kills the microbes in a very short time (28).

    The isolates of MRSA showed zero sensitivity to ampicillin; this effect did not show synergism when combined with Ag-NPs. Generally, the results of the antibiotic-Ag-NPs combination studies reported that Ag-NPs could enhance the antimicrobial activity of some antibiotics. However, many mechanisms of action were suggested for Ag-NPs antimicrobial activity such synergistic effect may not be completely clear (30). Also, the strong antimicrobial effect of Ag-NPs is due to attachment to cell receptors and inhibition of important metabolic enzymes resulting in disruption of microbial cell reproduction and respiration. Our results revealed that the killing rate was not concentration-dependent (31). Long-term cultivation enables bacterial cells to adhere to animal tissues and surfaces (32). This, in turn, results in the biofilm formation, a multilayered community of sessile bacterial cells. Bacteria growing in a biofilm are very problematic in hospitals and healthcare facilities due to their high antimicrobial resistance than planktonic cells. According to the National Institutes of Health, 80% of chronic infections are due to biofilms (33).

    In this study combinations of Ag-NPs and antibiotics also led to an increase in inhibitory effect on biofilm formation with different degrees, yet, generally showed a greater inhibitory activity than Ag-NPs or antibiotic alone. These are encouraging results, as it may be possible to achieve an effective anti-biofilm effect at lower antibiotic concentrations. Therefore, Ag-NPs and their combination with antibiotics could be used as an anti-biofilm agent against bacterial biofilms. From our study findings, it could be concluded that Ag-NPs have high antibacterial activity against MRSA, and could be an alternative to antibiotics for treatment of drug-resistant microorganisms specially MRSA infection, or enhancement of conventional antibiotics activity against MRSA and other pathogenic microorganisms. Further studies are required to understand the synergistic effect of nanosilver combination, and assess the safety and efficacy of new antibiotic- Ag-NPs combinations.
    Go to:
    Acknowledgments

    This project was supported by King Saud University, Deanship of Scientific Research, College of Science Research Center.
    Go to:
    Footnotes

    Authors’ Contribution:Ahmed Mohamed Ali Mohamed collected the samples and performed the practical work. Khalid A. Ali Abdel Rahim supervised, wrote and analyzed the data, and submitted this work.

    Funding/Support:This project was supported by King Saud University, Deanship of Scientific Research, College of Science Research Center.

    Go to:
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    Articles from Jundishapur Journal of Microbiology are provided here courtesy of Kowsar Medical Institute
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