Wednesday, January 30, 2019

Antibiotic collateral sensitivity is contingent on the repeatability of evolution

You know those moments when you read a paper and your head just explodes?  It's happened a few times to me in my life, and when it does the whole moment gets seared into my head.  One of those moments happened on a spring day in Oxford. I was eating a bap I had bought at the Taylor's expansion (1) by the old math's institute on Little Clarendon, sitting by a war monument (2). Here:


What I read was a paper from Dan Weinreich called Darwinian Evolution Can Follow Only Very Few Mutational Paths to Fitter Proteins. In this paper they engineering 2^5 = 32 strains of E. coli with 5 different basepair substitutions in a gene that encodes resistance to a certain kind of antibiotics. Then they simply measured the fitness of each of the strains, and constructed a hypercube, like Sewall Wright suggested first in 1932 (before anyone knew anything about genes!  he just called them allelomorphs).  If you make a 5-cube, like Wright sketched:

If every mutation gives you a fitness boost (as the literature showed they individually do) you should be able to get from wildtype (far left) to 'fully mutated' (far right) in 120 different ways. This would be something called a Mt. Fuji landscape, with one peak.  All paths go 'up' - which is essentially the null hypothesis that this paper was testing. At the time, I hadn't thought much about any other possible concepts -- being taught from the clinical oncology more-mutations-is-bad canon -- the more mutations a cancer has, the worse (more fit) it is.  (n.b. we can talk a lot about different 'kinds' of mutations - like passengers vs drivers, some other time, and also there's the whole fact that mutational effects are likely context dependent -- but we'll get to that later). Anyways, this is where the hair on fire part came for me. After they measure all the corner's fitnesses, they found that only 18 paths existed to get from left to right - and showed this figure:



There I sat, with my hair on fire, and a bap dangling out of my mouth. My world had changed forever.  What they showed here was empirical proof that the theoretical possibility that some combinations of mutations (even if they are individually beneficial) can be deleterious!  That means to two beneficial mutations could add up to a fitness penalty. This is something called epistasis (actually it's a special type called reciprocal sign epistasis, which you can read more about here).  While this was known to other members of the scientific community, it was not to me, and this set of a bunch of thinking that led to the idea that Dan Nichol and I (and friends) explored in our 2015 paper Steering Evolution with Sequential Therapy to Prevent the Emergence of Bacterial Antibiotic Resistance. Here, we used a set of fitness landscapes that were measured in a similar way to the above description, but for a lot of drugs, that were published by Mira et al. together with a mathematical model that Dan came up with.

This model is a time homogeneous absorbing Markov chain model of evolution that requires assumptions about the population all being on a single corner of the hypercube at any given time (often referred to as Strong Selection Weak Mutation (SSWM)).  Dan then calculated the probability of any given mutation from corner i to corner j ($P_{ij}$) based on changes in fitness from one corner to the next, like this:
Using this, and some matrix multiplication, we came up with the cute idea that evolution doesn't (necessarily) commute - that is, the evolutionary outcome could be quite different if you applied drug A and then B, as opposed to drug B and then A.

Given that many of the landscapes have more than one peak, there are also a number of situations in which evolution has to make a choice... it can go uphill in more than one direction. You can visualize this like a series of hills in which the population is 'walking up' them.  Given that the population can't 'see ahead' but can only make decisions about going 'up' instead of 'down', you can imagine that a population could easily evolve to an optima that is not globally optimal. You can see an example below (in panel (a)) where a strain starting at the yellow circle would move uphill to the blue triangle and then have to choose 'left' or 'right'. (obviously the geometry of this is wrong, but it is the best we can do as humans to visualize)


A few things become clear - one is that the fitness through time would be different (as you travel different trajectories even going 'up', it could be different along the way), and the other is that the end fitnesses will also differ (if you believe that evolution will EVER find a peak, which Artem doesn't -- more here in his bioRxiv preprint or on his blog).  AND if you then consider the end position, and how it relates to fitness IN ANOTHER LANDSCAPE (see panel (b), above for a visual representation), you have a situation where the outcome of drug sequencing can change with the evolutionary 'decision' - or in our jargon collateral sensitivity changing depending on evolutionary contingencies.

This led us to want to do the experiments. So, we continued our collaboration with Robert Bonomo at the Cleveland VA hospital, and performed 60 replicates of the same experiment - 60 what we now term evolutionary replicates. Just showing the first 12, we see evidence for the different trajectories encoding different fitnesses through time, here:

Out of curiosity, we wondered how common it would be to get a different answer to the question: If I give one drug (drug A) and then another (drug B), how much variation could I see in collateral response. To answer this, we used another version of the mathematical model from our first paper (which you can download here, along with the data needed to redo our analysis). It turns out that there is wide variability in collateral sensitivity (according to our mathematical model), so much that any one repeat of an evolution experiment could give you an opposite answer (could reveal collateral sensitivity) when the very next one could reveal cross-resistance.


This is sorta bad news...  but to try to find the bright side, we propose using a new metric, which we call Collateral Sensitivity Likelihood (CSL), which is a measure of a sequence of drugs providing any collateral sensitivity at all.  This would make for safer recommendations -- where what you want is some method for clinicians to rationally choose a drug ordering that has a high probability of being better (or the opposite, a low probability of the drugs inducing resistance to one another).

So - from hair on fire to 5 years of research later, we finally were able to get this story out there. We published it last week, and you can read the whole paper, which is open access, here: Antibiotic collateral sensitivity is contingent on the repeatability of evolution.  Lots more details and figures are available there...

In addition to making the code and phenotype data available (in the embedded github repo link), we also performed whole genome sequencing on 12 of the evolutionary replicates, and we uploaded those data to the NCBI Sequence Read Archive (SRA), and they are freely available through accession code PRJNA515080, or through this link. So, hurray for #openscience - we'd love any secondary analyses or ideas for future projects.  Lots to think about.

If this research interests you, please check out our lab page to see what else we're up to.  We're trying to apply evolutionary thinking, mathematical models and experimental evolution to cancer and pathogens to ease suffering.

61 comments:

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    He can still be able to help you with this herbs medicine:
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  5. Joy and happiness is all i see now I am here today to appreciate the good
    work of Dr Ohunyom, for helping me curing my HSV 1&2 Disease, I contacted
    the virus from my Partner, i found out he was cheating on me, so i broke up
    with him, 2 month later i went to test myself in the hospital when i was
    seeing sore on my mouth and Private areas, i was diagnosed of HSV 1&2. i
    tried using the acyclovir medicine but it didn't help in curing it, i went
    on a search and found out Dr Ohunyom curing people illness, and so i got
    his email and i was told how i can get his herbal cure, 5 days later i
    received his medicine after using it for 2 weeks i was tested again and was
    confirm negative, today i am totally free. If you are over there looking
    for solution email drohunyom@gmail.com or contact him on whatsapp
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    cure for the Following DISEASES, eczema,urethra wart,chronic
    problems.Herpes, Cancer, Als,Hepatitis, Diabetes, HPV,Infections,ulcer ETC

    ReplyDelete
  6. i already gave up on ever getting cured of HSV2 because i have try many treatment none of them work out for me i have go to different hospital they always tell me same thing there is no cure for herpes when i came across a post about Dr ohunyom, in the net from a lady called Angela i contacted him and he reassured me with him herbal medicine which i took according to the way he instructed, that how i was cured. I doubted at first because i have been to a whole lot of reputable doctors, tried a lot of medicines but none was able to cure me. so i decided to listen to him and he commenced treatment, and under two weeks i was totally fee from #Herpes. i want to say a very big thank you to DR ohunyom for what he has done in my life. feel free to leave him a message on email drohunyom@gmail.com and also WhatsApp him +2349060579973..
    He can still be able to help you with this herbs medicine:
    1...ALS CURE/DIABETES CURE/EPILESY/HPV CURE/LUPUS/HEPATITIS/CANCER/GOUT    

    ReplyDelete
  7. Happiness is all i see now I never thought that I will be cured from HERPES virus again. I have been suffering from a deadly disease (HERPES) for the past 3 years now, I had spent a lot of money going from one places to another, from churches to churches, hospitals have been my home every day residence. Constant checks up have been my hobby not until this faithful day, I was searching through the internet, I saw a testimony on how DR Ohunyom helped someone in curing his HERPES disease, quickly I copied his email which is
    drohunyom@gmail.com just to give him a try I spoke to him, he asked me to do some certain things which I did, he told me that he is going to provide the herbal cure to me, which he did, then he asked me to go for medical checkup after some days after using the herbal cure, behold I was free from the deadly disease, he only asked me to post the testimony through the whole world, faithfully am doing it now, please brothers and sisters, he is great, I owe him in return. if you are having a similar problem just email him on (drohunyom@gmail.com) or you can whatsApp him +2349060579973 and He also has herbal cure for the Following DISEASES, eczema,urethra wart,chronic problems.Herpes, Cancer, Als,Hepatitis, Diabetes, HPV,Infections,ulcer ETC

    ReplyDelete
    Replies
    1. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line:+2348101755322 or his Email:Oyaguspellcaster@gmail.com

      Delete
    2. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line:+2348101755322 or his Email:Oyaguspellcaster@gmail.com

      Delete

  8. I want to use the means to thank Robinson.buckler for his good work I have been very sick with this horrible diseases for 9 months they all tell me there was no cure for me I have go to different hospital I always have same news, I was confused because I feel like ending my life that time thank God I was able to locate doctor Robinson.buckler who help me I found his email on site how people was writing Good about him I never believe he can really cured me until when I get my test done. I am so happy to tell you this Robinson.buckler have a herbal cure to Herpes and you can take the medicine according to his instruction you will found out that you will give testimony like me I take his herbal medicine for just 2 week. I am so happy today that I am really free from this sickness here is Robinson.buckler email Robinson.buckler (( @yahoo. com ))………………………………. you can reach him out now https://robinbuckler.com/

    ReplyDelete
  9. 5 years ago I had warts, I was treated with some liquid applied to the warts they continued to grow and spread... The next 2 doctors did laser surgery to remove them. 1 year after the surgery, they grew back close to where the 1st ones were' so I was finally told it was hpv. I have had it for very long time, I contract it from my cheated boyfriend and I found out he was also infected and I end up the relationship between us. the warts was so embarrasses because it started spreading all over I have be dealing with this things for very long time the last treatment I take was About 2 years ago I applied natural treatment from Dr onokun herbal cure, a week after applying the treatment all the warts was gone. it's now 2 years and some months I don't have single wart or any symptoms of hpv. wow"" it's great, Dr onokun has finally cured me. Anyone living with hpv contact Dr onokun for natural treatment.
    His email address: dronokunherbalcure@gm‎ail.com  

    ReplyDelete
  10. This has really worked and I am proud to testify of it. I saw a post on how a lady got her man back by Dr Jato. He helped her to get her Ex Husband back. My Husband divorce me 2 years ago to live with another woman and i have try to let go of him but i could not, so i try to do many things to get him back and he refuse to come back to me. I visit a forum one day for tips to get him back and there is saw a lady testifying of how Dr Jato helped he to get her man back after he was tie down with a spell by another woman and Dr Jato was the one that help her. Although I never believed in spiritual work I reluctantly tried him because I was desperate and i contact him and explain my problems to him, to my greatest surprise Dr Jato helped me to bring back my man after 2 days and now my relationship is now perfect just as he promised. Getting your ex back permanently spell does not only bring back someone you love but it will also re-ignite your lovers feelings for you to be as happy with that person. My man now treats me like a queen and always say he love me all the time. If you are passing through difficulties in your relationship Email him for urgent help.E-mail: jatolovespell@gmail.com or WhatsApp/Call him at +2348140033827
    thank you Dr Jato for the help...

    ReplyDelete
  11. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line:+2348101755322 or his Email:Oyaguspellcaster@gmail.com

    ReplyDelete
  12. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line:+2348101755322 or his Email:Oyaguspellcaster@gmail.com

    ReplyDelete
  13. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line:+2348101755322 or his Email:Oyaguspellcaster@gmail.com

    ReplyDelete
  14. I have come to find out that herbal medicine its the best solution to all STDs and STIs and other health related threatening issues/diseases, cause i was once tested positive for herpes and i have lived with it for about 5 years tried all form of medication all to know avail, until i get a recommendation from someone online about herbal medicine fro Dr Ofua, indeed i actually do a little/more research about the Herbalist and found out he has the possible cure to my ailment, i gave it a try by contacting him on WhatsApp messenger +2347059497969 he had the herbal medicine he sent to me and gave me dietary instructions to follows up while using the herbal medicine, he suggest that i used the medicine for a period of 21 days that after then i should go for a check up at the hospital to know the status of my herpes, which i actually did, after few weeks as recommended by him, I go for check up and behold i was now tested Negative, ever since then i have vowed to let everyone knows that herbal medicine is the solution to any diseases/infections, despite these Big pharma company trying to makes us not to use herbal medicine framing all sort of bad reviews just to diminished herbal medication for their selfish interest and big wallets purpose, my advice to you all is that try herbal medicine today and be glad that you have tried it and i assured you that if you do you will have cause to share a testimony online just am doing now, visit (https://the-great-herbal-healing-home.business.site) or email thegreatherbalhealinghome@gmaill.com to know more about the great herbalist that help me and am sure he too will help you.

    ReplyDelete
  15. I am here to give my testimony about chief dr lucky who helped me in my life, i want to inform the public how i was cured from herpes simplex virus by chief dr lucky, i visited different hospital but they gave me list of drugs like Famvir, Zovirax, and Valtrex which is very expensive to treat the symptoms and never cured me. I was browsing through the Internet searching for remedy on herpes and i saw comment of people talking about how chief dr lucky cured them. when i contacted him he gave me hope and send a Herbal medicine to me that i took and it seriously worked for me, am a free person now without problem, my herpes result came out negative. I pray for you chief dr lucky am cured you can also get your self cured my friends if you really need my doctor help, you can reach him now.
    Email: chiefdrlucky@gmail.com  Call or What's App +2348132777335  
    Website :  http://chiefdrluckysolutionhome.website2.me/ 

    ReplyDelete
  16. Hello all viewer online HIV has cure but doctor said Hiv/Aids has no cure until i met Dr SAYO who help me in my life. I was infected with HIV/AIDS in 2008, i went to many hospitals for cure but there was no solution, so I was thinking how can I get a solution, so that my body can be okay. until this faithful day as i was browsing the net I saw a testimony on how Dr.SAYO helped people in curing HIV Disease, quickly I copied his Email which is sayoherbalhealer@gmail.com so i contacted him for solution for my HIV, so Dr.SAYO told me that his going to prepare herbal medicine for my health, then he prepared the medicine and luckily after two week my Hiv/Aids was be cured. Dr SAYO is well recognize as one of the best herbalist doctor in Africa, you don't have to be sad anymore or share your tears anymore on this disease when the cure have already be found by Dr.SAYO herbal medicine, he so cure HERPES,CANCER, ALS,HEPATITIS B, DIABETIC check his FB page:  https://www.facebook.com/Sayo-Herbal-Healer-100145798345000/ OR visit his websites website; http://sayoherbalhealer.website2.me/  check blog: https://sayoherbalhealer.blogspot.com/     

    ReplyDelete
  17. Greatest thanks to Dr Oyagu for his herbal drugs that he prepared for me and when i start using it in just 2weeks i was completely cured and that ended my HERPES SIMPLEX 1&2 DISEASE i am so happy and grateful to Dr Oyagu . after reading about him on a testimony of Jason Cash on a blogger. i knew suddenly Dr Oyagu was the right Doctor to cure my HERPES SIMPLEX 1&2 DISEASE. i discuss with Dr Oyagu and he prepared a herbal medicine for me and when it got sent to me in south korean . i used the herbal medicine and 2weeks and i went to check up again. after 15years of suffering from HERPES SIMPLEX 1&2 at last i am smiling once again. Dr Oyagu also has remedy to others disease like COLD SORES,HIV/AIDS,DIABETES.CANCER,HIGH BLOOD PRESSURE AND MANY MORE. I oblige everyone to contact this powerful herbalist Dr Oyagu and be free from your suffering. contact his WhatsApp line: +2348101755322 or his Email:Oyaguherbalhome@gmail.com

    ReplyDelete
  18. I  got diagnosed with HSV 1, and reading this actually made me feel like crawling and crying like a baby. The outbreak I have right now is genital and in my throat and mouth, I talked to the doctor who did the blood test on me and said over time that it would eventually get much better. She said that what I’m having is a massive outbreak and is unusual, but that it may be due to the fact that I stressed myself out over it so much. She also told me even though I have it in the genital area that It doesn't mean that I needed “private to private” contact to acquire it. But this did make me feel a whole lot better about the whole situation. For anyone else to have HSV 1 after all this stress and pains am passing and the doctor saying all such of crap i was not convince by his Theory so i decided to go on the internet and saw this comments by Rose who testify about how she get cured by root and herbs by Dr KHAM, when i first bump into it i didn't believe the comments at first so i decided to confront her on Facebook and really know what and how did it work because my doctor told me there is no cure for herpes, so i really contacted DR KHAM, he responded in A minute the only thing he ever ask for was just the items of which he will use to prepare for the herbs, after all this he send the herbs through courier Services, within a week i was cure just like that without no struggle or stress, if you are out there in need of cure contact him with his email address: dr.khamcaregiver@gmail.com or dr.khamcaregiver@outlook.com   OR WHATSAPP HIM ON +2348159922297,  You can Also Visit his website to know more about him at > https://drkhamherbalhealingcenter.wordpress.com/     

    ReplyDelete
  19. Hello everyone i am happy to say that i was cure from HIV/AIDS by Dr Padman with his herbal medications after i tested positive july 2016 and ever since then i have been on medications to extend my living on earth. One faithful day i was searching on line if i could get some tips on how to cure my illness and i saw a comment on how Dr Padman cure HIV with herbs,i was surprise and i contact him on the email they provided on the comment and i explain my problem to him and he also prepare the herbs and send to me which i use for one week as i was instructed by him and after 2weeks i went to hospital for checkup and i tested Negative and i am fully cured from the disease and i am so greatful to Dr Padman.You can also reach Dr Padman on his padmanlovespell@yahoo.com , https://twitter.com/padman_dr. https://padmanspell.com WhatsApp +19492293867

    ReplyDelete
  20. HOW DR PERFECT ENLARGEMENT CREAM HELP ME TO ENLARGE MY PENIS YOU CAN ADD HIM UP ON WHATSAPP +1(256) 251 3614

    My name is David Richardson from USA I want to share a testimony of how Dr Perfect herbal mixture cream saved me from shame and disgrace, my penis was a big problem to me as the size was really so embarrassing and I was also having weak erection problem. I can't make love to my wife and my penis was just too small, a full grown man like me having 3 inches penis and to worsen it I don't last long in sex I can't even last four minutes it was really a big shame to me. My wife was really tired of me because my sex life was very poor, she never enjoyed sex, I was always thinking and searching for solutions everywhere until when I saw a testimony of how Dr Perfect herbal mixture cream have been helping people regarding their sex life, so I decided to give him a try and to my greatest surprise in less than one week of taking the herbs my penis grow to 10 inches I couldn't believe my eyes and as I speak now my penis is now 10 inches long and I don't have weak erection anymore. I can make love to my wife longer in bed. And my marriage is now stable my wife is now enjoy me very well in bed. All thanks to Dr Perfect for enlarging my penis he indeed save my marriage. You can contact him at solutiontoallproblems1@gmail.com or you can add him up on WhatsApp +1(256) 251 3614. you can also view my blog http://davidrichardson12356.over-blog.com. He also specializes in the following things
    Herpes virus 
    Cancer
    Hsv
    Chronic venous disease
    and others 

    ReplyDelete
  21. HOW DR PERFECT ENLARGEMENT CREAM HELP ME TO ENLARGE MY PENIS YOU CAN ADD HIM UP ON WHATSAPP +1(256) 251 3614

    My name is David Richardson from USA I want to share a testimony of how Dr Perfect herbal mixture cream saved me from shame and disgrace, my penis was a big problem to me as the size was really so embarrassing and I was also having weak erection problem. I can't make love to my wife and my penis was just too small, a full grown man like me having 3 inches penis and to worsen it I don't last long in sex I can't even last four minutes it was really a big shame to me. My wife was really tired of me because my sex life was very poor, she never enjoyed sex, I was always thinking and searching for solutions everywhere until when I saw a testimony of how Dr Perfect herbal mixture cream have been helping people regarding their sex life, so I decided to give him a try and to my greatest surprise in less than one week of taking the herbs my penis grow to 10 inches I couldn't believe my eyes and as I speak now my penis is now 10 inches long and I don't have weak erection anymore. I can make love to my wife longer in bed. And my marriage is now stable my wife is now enjoy me very well in bed. All thanks to Dr Perfect for enlarging my penis he indeed save my marriage. You can contact him at solutiontoallproblems1@gmail.com or you can add him up on WhatsApp +1(256) 251 3614. you can also view my blog http://davidrichardson12356.over-blog.com. He also specializes in the following things
    Herpes virus 
    Cancer
    Hsv
    Chronic venous disease
    and others 

    ReplyDelete
  22. I have read so much on Herpes Simplex virus. I hear of reaching clearance, I hear of no cure, I hear of recurrent lesions, poor immune system, poor diet, etc. I just do not know what to believe. I have visited many web pages that seem to sell you their version of removing or enforcing the immune system. Then I hear all the info on therapeutic vaccines as well as prophylactic vaccines and wonder how can that even help me. And I feel so worthless, Glad i came across Doctor Razor article on how to heal HSV forever. Endless Gratitude to doctor Razor for helping me get rid of my Herpes Virus. Reach him on his clinic email drrazorherbalhome@gmail.com
    Whatapp His cell phone on +2349065420442

    ReplyDelete
  23. I have read so much on Herpes Simplex virus. I hear of reaching clearance, I hear of no cure, I hear of recurrent lesions, poor immune system, poor diet, etc. I just do not know what to believe. I have visited many web pages that seem to sell you their version of removing or enforcing the immune system. Then I hear all the info on therapeutic vaccines as well as prophylactic vaccines and wonder how can that even help me. And I feel so worthless, Glad i came across Doctor Razor article on how to heal HSV forever. Endless Gratitude to doctor Razor for helping me get rid of my Herpes Virus. Reach him on his clinic email drrazorherbalhome@gmail.com
    Whatapp His cell phone on +2349065420442

    ReplyDelete
  24. I have read so much on Herpes Simplex virus. I hear of reaching clearance, I hear of no cure, I hear of recurrent lesions, poor immune system, poor diet, etc. I just do not know what to believe. I have visited many web pages that seem to sell you their version of removing or enforcing the immune system. Then I hear all the info on therapeutic vaccines as well as prophylactic vaccines and wonder how can that even help me. And I feel so worthless, Glad i came across Doctor Razor article on how to heal HSV forever. Endless Gratitude to doctor Razor for helping me get rid of my Herpes Virus. Reach him on his clinic email drrazorherbalhome@gmail.com
    Whatapp His cell phone on +2349065420442

    ReplyDelete
  25. Its a miracle that I am alive today. I encounter HERPES SIMPLEX VIRUS (HSV1&2) for good four (4) years, six (6) months and Ten (10) days with my calculations, before I met Doctor Razor. I am from New York City ,NY USA,I was affected with herpes simplex virus.  Herpes is a viral infection cause by the Herpes simplex virus, marked by painful, watery blister in the skin or mucus membrane or on the genitals. This was what I suffered for over 4 years. I have visited so many hospitals in Houston for treatment but no improvement. I met Doctor Razor on the internet while I was searching for herpes cure,I read some testimonies about his work. I decided to give  his herbal medication a trial, which have now become a testimony in my life. i am so grateful to God almighty for sending doctor Razor my way when i thought all hope was lost. You too can get cured of herpes Simplex virus, if i could overcome it. Reach out to doctor razor via his clinic email : drrazorherbalhome@gmail.com. Whatsapp his cell phone ON +2349065420442

    ReplyDelete
  26. Its a miracle that I am alive today. I encounter HERPES SIMPLEX VIRUS (HSV1&2) for good four (4) years, six (6) months and Ten (10) days with my calculations, before I met Doctor Razor. I am from New York City ,NY USA,I was affected with herpes simplex virus.  Herpes is a viral infection cause by the Herpes simplex virus, marked by painful, watery blister in the skin or mucus membrane or on the genitals. This was what I suffered for over 4 years. I have visited so many hospitals in Houston for treatment but no improvement. I met Doctor Razor on the internet while I was searching for herpes cure,I read some testimonies about his work. I decided to give  his herbal medication a trial, which have now become a testimony in my life. i am so grateful to God almighty for sending doctor Razor my way when i thought all hope was lost. You too can get cured of herpes Simplex virus, if i could overcome it. Reach out to doctor razor via his clinic email : drrazorherbalhome@gmail.com. Whatsapp his cell phone ON +2349065420442

    ReplyDelete
  27. A MIRACLE TESTIMONY FOR WHAT Dr SAY0 HAS DONE FOR ME!!  
    Human PapilomaVirus (HPV) Cure Found By Dr Sayo If you're wondering why am here or why am doing this!, i just want to show appreciation to God, and i want other persons who is suffering from this deadly illness, ( Herpes, Coronary Artery Disease (Ischemic Heart Disease), Hiv/AID, Stroke, Trachea, Bronchus, and Lung Cancers, Diarrhea Diseases, Diabetes Mellitus, Dengue, Rota-virus, Other STD and others ) maybe because of ignorance. if you see this don't ignore, i was a victim of hpv, some one introduced him to me, i wrote him, told him what i was going through,  i was not too sure he could help, because i seen other persons, or post but no good outcome, he told me he could help, like others, i got the medicine or the herb whatever you may want to call it, he didn't even charge me a lot, to God i didn't want to use it, but i did, to God be the glory am the one sending this post today in good health and sound mind, ..smile.. i seek for him, i didn't mind the distance so i could show appreciation, i meet with Dr Sayo he didn't ask for anything, all he said is, 'It's God doing. I saw him treat people with different situation and problem maybe worse than yours, tell others. so am telling you today, don't ignore this, if you see this post, tell the next person who you think might need this. contact him,on email (sayoherbalhealer@gmail.com) check his FB page:  https://www.facebook.com/Sayo-Herbal-Healer-100145798345000/ OR visit his websites website; http://sayoherbalhealer.website2.me/  check blog: https://sayoherbalhealer.blogspot.com/

    ReplyDelete
  28. HOW I WAS ABLE TO FLUSH OUT THE HERPES VIRUS WITH THE HELP OF DR. KHAM NATURAL PRODUCT, HIS EMAIL dr.khamcaregiver@gmail.com. I was diagnosed with Herpes a year ago. It’s the worst thing that’s ever happened to me and  I feel like my life is over. Living with constant pain is something no one should have to go through, Herpes has also had a huge impact on my sleep . Since being diagnosed I’ve fallen so far into depression I see no way out because i have been to different hospital. I’m taking a high dose of antidepressants, as well as sedatives and sleeping-pills. I don’t know how I’ll ever accept that this is my life now…I keep asking why me because know changes? But I guess reading your testimony really made me feel happy because i believe what i had from people out there who you also help to cure the herpes virus, and i believe so much with your herbal medicine i am going to be out of this bad situation like others you also help before, what we i have done if not for the herbal medicine that i requested for that was able to flush out the bad virus that was making me go crazy. people out there how will i thank this man called DR. KHAM, i will advice you to email him direct if you have disease like herpes etc. dr.khamcaregiver@gmail.com or whatsapp him +2348159922297

    ReplyDelete
  29. I want to thank Dr Emu a very powerful spell caster who help me to bring my husband back to me, few month ago i have a serious problem with my husband, to the extend that he left the house, and he started dating another woman and he stayed with the woman, i tried all i can to bring him back, but all my effort was useless until the day my friend came to my house and i told her every thing that had happened between me and my husband, then she told me of a powerful spell caster who help her when she was in the same problem I then contact Dr Emu and told him every thing and he told me not to worry my self again that my husband will come back to me after he has cast a spell on him, i thought it was a joke, after he had finish casting the spell, he told me that he had just finish casting the spell, to my greatest surprise within 48 hours, my husband really came back begging me to forgive him, if you need his help you can contact him with via email: Emutemple@gmail.com or add him up on his whatsapp +2347012841542 is willing to help any body that need his help.
    Website: http://emutemple.website2.me/

    ReplyDelete
  30. We all can help him share his great work so order people can see it plz OK., Dr Oyakhire has really made me so much believe in him by getting me cure with his herbal treatment. i really appreciate you Dr Oyakhire for bringing back happiness to my life again. thanks you so much friends that join me to thank Dr.Oyakhire for what he has actually done for me i pray to you all a good life and properity, email him at ...DrOyakhire7@gmail.com
    Whatsapp him +2349039208180
    He did cure those illness with herbal medicine!!!

    1 Cancer herbal medicine

    2 Hiv herbal medicine

    3 Low sperm count

    4 Barrenness

    5 Hvp herbal medicine

    6 Herpes herbal medicine

    7 Diabetes

    8 Love spell.

    9 Ex back

    Thanks Dr Oyakhire for your help in my life and family... and most especially to you Dr Oyakhire THANKS

    ReplyDelete

  31. I came on the Internet i saw great testimony about DR Voke on how he was able to cure someone from HERPES, and any disease this person said great things about this man, and advice we contact him for any problem that DR Voke can be of help, well i decided to give him a try, he requested for my information which i sent to him, and he told me he was going to prepare for me a healing portion, which he wanted me to take for days, and after which i should go back to the hospital for check up, well after taking all the treatment sent to me by DR Voke i went back to the Hospital for check up, and now i have been confirmed HERPES Negative, friends you can reach Dr voke on any treatment for any Disease, reach him on _________________________________doctorvoke@gmail. com

    ReplyDelete

  32. I had my first seizure. I was in high school and didn't want anyone to know that I had epilepsy. I had seizures on two occasions. I was so ashamed and didn't want people to think that something was wrong with me. I finally got to the point where I told people that I had epilepsy. I came to the place that I realized that I had to own my epilepsy or it would control me. Epilepsy hadn't really affected my life until about 9 years ago after my first child was born... I am a wife and mother of two beautiful children. I have lived with epilepsy for 20 years Yet, my life has been dramatically impacted by epilepsy. I had search for cure online without no hope. I met a Man called DR Lewis hill, i contact him I made an order of his medicine and I do not experience seizure anymore i was cured form it. So if you are in the same situation as mine contact him on this email drlewishill247@gmail.com

    He cure listed diseases

    .CANCER
    .HIV
    .ALS
    .HEPATITIS A AND B
    .DIABETIC.
    .HERPES

    ReplyDelete
  33. I'm here to testify about the great work Dr Odudu  did for me. I have been suffering from (HERPES) disease for the past 4 years and had constant pain, especially in my knees. During the first year,I had faith in God that i would be healed someday.This disease started circulating all over my body and i have been taking treatment from my doctor, few weeks ago i came across a testimony of one lady on the internet testifying about a Man called Dr Odudu on how he cured her from Herpes Simplex Virus. And she also gave the email address of this man and advise anybody to contact him for help for any kind of sickness that he would be of help, so I emailed him telling him about my (HERPES Virus) he told me not to worry that i was going to be cured!! Well i never believed it,, well after all the procedures and remedy given to me by this man few weeks later i started experiencing changes all over me as Dr Odudu assured me that i will be cured,after some time i went to my doctor to confirmed if i have be finally healed behold it was TRUE, So friends my advise is if you have such sickness or any other at all you can contact Dr Odudu via email dr.oduduhome@gmail.com You can also call or whatsApp his telephone number on +2347041800280 Thanks  once again Dr Odudu.

    ReplyDelete
  34. Dr. Kham herbal medicine is the best remedy for Herpes and HIV/AIDS, I was a carrier of Herpes and I saw a testimony on how Dr. Kham cure Herpes, I decided to have a contact with him and asked him for help and he started the remedies for my health and prepare the herbal medicine for me which i use to cure myself. Thank God, now everything is fine, I’m cured by Dr. Kham herbal medicine, I’m very thankful to Dr.Kham and i will not stop publishing his name on the internet because of the good work he did for me, You can reach him on his email: dr.khamcaregiver@gmail.com Or you can call and WhatsApp him on +2348159922297, You can also visit his website: https://drkhamherbalhealingcenter.wordpress.com/    


    DOCTOR KHAM CAN AS WELL CURE THE FOLLOWING DISEASE:-

    1. HIV/AIDS
    2. HERPES
    3. CANCER OF ALL KINDS
    4. HSV
    5. Hepatitis A,B,C
    6. Diabetes
    7. CHRONIC DISEASE

    Natural remedy for curing HERPES and HIV. Call and whatsapp Dr Kham on +2348159922297  

    ReplyDelete
  35. I have been suffering from Herpes for the past 3 years and 8 months, and ever since then i have been taking series of treatment but there was no improvement until i came across testimonies of Dr Sayo on how he has been curing different people from different diseases all over the world, then i contacted him as well. After our conversation he sent me the medicine which i took according to his instructions. When i was done taking the herbal medicine i went for a medical checkup and to my greatest surprise i was cured from Herpes. My heart is so filled with joy. If you are suffering from Herpes or any other disease you can contact Dr Sayo today on this Email address: (sayoherbalhealer@gmail.com) or WhatsApp him on this Number +2349012175679 Visit his FB page: https://www.facebook.com/Sayo-Herbal-Healer-100145798345000/    also visit website: http://sayoherbalhealinghome.com/ or check blogs: https://naturerootandherbscures.blogspot.com/

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  36. NATURAL HERBS  AND REMEDIES FOR CURING HIV, HERPES, HPV, HSV1&2, CANCER AND DIABETES I just experienced the wonders of Dr. Kham herbs. God will continue to bless you Dr. Kham more abundantly for the good work you are doing in people's life by curing them from different types of diseases. I will keep on testifying about your good work. I was diagnosed with herpes 6 months ago and my doctor told me that there is no cure. I did not believe him and I keep my faith up hoping that one day I will be cured from this horrible disease. One day I saw a post on the Herpes Virus Blog forum about a herbal doctor called Dr. Kham on how he cures people from diseases with his herbs and I quickly contacted Dr. Kham on his email explained my problem to him and he told me not to worry that he is going to cure me and truly he prepared the cure and sent it to me through UPS and gave me instructions on how to take it. After two weeks of taking the herbal Medication, I went to my doctor for a checkup and my results showed Negative and my doctor told me that there is no trace of herpes in my body and the virus is completely gone from my body. Everybody at the clinic rejoices with me and they all ask for Dr. Kham contact. You can contact Dr. Kham on his email: dr.khamcaregiver@gmail.com or add whats-app +2348159922297  and you can get in touch with him on his website: https://drkhamherbalhealingcenter.wordpress.com/   or https://drkhamcaregiver.wixsite.com/drkhamcaregiverherba , He also have herbs medicine to cured the following diseases: [eczema], [urethra wart], [chronic problems], [Herpes], [Cancer], [Als], [Hepatitis], [Diabetes], [HPV], [Infertility/PCOS Infections] , [ulcer  ETC......

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  37. I was cured of CANCER with the used of natural herbs. My name is Mella Ken and am from US. I love herbs so much. Most times, injection and drugs are just a waste of time. I was cured 8 months ago, i suffered from CANCER for 3 yrs but with the help of Dr.Sayo herbal medicine, i was cured within few weeks of drinking the herbs he sent to me through courier delivery service. This same doctor also cured my Aunty from FIBROID, as soon as i heard she had FIBROID, i quickly refer her to Dr.Sayo and she was cured too after drinking his herbs.I have referred more than 15 persons to Dr.Sayo and they were all cured from their various illness. Have you taken herbs before?. You have spent so much money on drugs,injections,surgeries etc and yet you have no good result to show for it. Contact Dr. Sayo now, he is a herbalist doctor, i assured you of a cure if you drink his natural herbs. Dr.Sayo have herbs that cures Hiv, Herpes, diabetics, asthma, hepatitis, HBP, STD, cancer, chronic, etc. Contact Dr Sayo through his Email address on: sayoherbalhealer@gmail.com or WHATSAPP/CALL him on+2349012175679 Visit his FB page: https://www.facebook.com/Sayo-Herbal-Healer-100145798345000/    
    also visit website: http://sayoherbalhealinghome.com/ or check blogs: https://naturerootandherbscures.blogspot.com/ Please share the good news to other people once you are cured    

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  38. I want to give a testimony on how I got cured from Herpes Virus. Few months back I was having some symptoms. I went to see a doctor and many blood tests was done on me, later on I was told I had Herpes. My doctor told me that there’s no cure for Herpes I felt bad, I went online searching for a possible cure for Herpes Virus, I saw a post of Dr.Ojamo a herbal doctor that’s helping people get cured from Herpes. I contacted him and told him how I’m feeling he said his herbal medicine can cure me. He sent me the medicine via UPS delivery service and I received the medicine in 8days after he sent it, i took the medicine as prescribed by him. Before the completion of the medication the symptoms stopped. I went to the doctor and carried out another blood test, surprisingly I was Negative. I haven’t had any symptoms anymore. Dr. Ojamo is great and you can contact him via dr.ojamoherbalhome@gmail.com or message him on via whatsApp at +2349077406037

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  39. Natural herbs have cured so many illness that drugs and injection cant cure. I've seen the great importance of natural herbs and the wonderful work they have done in people's lives. i read people's testimonies online on how they were cured of herpes, HIV, cancer, lutos diabetics etc by Dr. ISHIAKU  herbal medicine, so i decided to contact the doctor, I was diagnosed with herpes for the past 6 month but Dr. ISHIAKU cured me with his herbs and i referred my aunt and her husband to him immediately because they were both suffering from hiv but to God be the glory, they were cured too .I know is hard to believe but am a living testimony. There is no harm trying herbs. He is also a spell caster, he cast spell to restore broken marriages and he cast good luck spells to prosper and excel in life. Contact Dr ISHIAKU on Whatsapp or call him +2348180828544 OR email him ISHIAKUHERBALCURE@GMAIL.COM

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  40. I'M Maria Gomaez God bless Dr. Asein for his marvelous work in my life, I was diagnosed of HIV since 2018 and I was taking my medications, I wasn’t comfortable i needed to get the HIV out of my system, I searched about some possible cure for HIV i saw a comment about Dr. Asein, how he cured HIV with his herbal medicine, I contacted him and he guided me. I asked for solutions, he started the remedy for my health, he sent me the medicine through UPS COURIER DELIVERY SERVICE. I took the medicine as prescribed by him and 14 days later i was cured from HIV, Dr. Asein truly you are great, do you need his help also? Why don’t you contact him through dr.aseinherbalhome@gmail.com.....dr.aseinherbalhome@hotmail.com or whatsapp him on +2347059207062.he has cure for
    Herpes
    Hbv
    And kidney disease...

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  41. i want to tell the whole world of great Herbalist herbal mixture CREAM. i was unhappy ever since my girlfriend back then in college mocked me of having a small penis and weak in bed ,ever since then i have been trying to improve my sex life but all effort failed ..but it got worse when i got married ,my wife is so angry with me and Threatened to leave me if i do not find a way out and i love my wife so much, so one faithful day i was reading some comments on a blog then i saw a comment of someone testifying of how Herbalist Dr helped him enlarge his penis and Fertilize his sperm count so i quickly contacted him by his email and he got back to me and i did as he said ..to my greatest surprise i got 10 inches within two weeks of using his herbal mixture cream and now its just 3 weeks i am having 10.5 inches i am so happy now..my wife told me am so much good in bed now all thanks to OGODO herbal mixture cream for saving my marriage and disgrace. NOTE: he also cure sicknesses and diseases like
    PENIS ENLARGEMENT ERECTILE DYSFUNCTION DIABETES type 1 and type 2 HERPES GENITAL WART
    LOW SPERM COUNT WEAK ERECTION
    BREAST ENLARGEMENT
    PROSTRATE CANCER
    HIV/AIDS
    PREMATURE EJACULATION
    LOVE SPELL AND BRING YOUR EX BACK
    You can reach him via email OGODOHERBALHOMESOLUTION@GMAIL.COM or WHATAPP HIM +23490440680467 You Can check his website on his Facebook page https://www.facebook.com/oogoherbalhomesolution/

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  42. Before people said there is no cure for herpes virus but today many people have now believe that there is a cure,herpes virus can be cured through Africans roots and herbs, Dr.chala he is the one of the great herbal doctor in Africa and he has the cure on this virus last month he share his herbal medicine in some medical hospital and now he is well recognize as one of the best in Africa, you don’t have to be sad any more or share your tears any more on this virus when the cure have already be find by Dr.Chala email him on dr.chalaherbalhome@gmail.com or https://
    drchalaherbalhome.godaddysites.com or https://mywa.link/dr.chalaherbalhome

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  43. HERBAL DR EMU WHO PREPARE HERBAL MEDICINE TO CURE ALL KINDS OF DISEASES INCLUDING HERPES DISEASE.   
     I have been battling this Herpes disease for almost 3 years now....I tried all possible means to get cure from my Herpes Disease but all to be in vain until i saw a post in a health forum about a herbal Dr Emu who prepare herbal medicine to cure all kind of diseases including Herpes Disease, at first i doubted if it was real but decided to give it a try...when i contact Dr Emu via his email (emutemple@gmail.com)  write him and reply me explain how the process work so after ordering for the medicine I got it within 3/4 working days through DHL Delivery and I took it according to the way Dr Emu instructed, I was so happy after 2 week I took the medicine there was very big change in my health when I was done with the process I go for test, I found out I am negative...Herpes patients should also get in touch with this herbalist Dr Emu to get rid of these Herpes Virus forever his whatsapp number +2347012841542. Website: https://emutemple.wordpress.com/ 

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  44. I have been a herpes carrier for 2 years and I tried every possible means
    of curing but all of no useful until I saw a health promotion on a
    herbalist from Africa who prepares herbal medicines to cure all sorts of
    diseases including cancer and many others sickness, I first doubted It was
    not true but decided to try, when I contacted this herbal physician via his
    email, I purchased herpes herbal medicine and received it within 7 days
    and used it as prescribed, I tested negative within 3 weeks of use, do not
    lose hope to contact him early If you need help, am convinced he can help you too,
    He is also special in curing 1.
    HIV/AIDS 2. HERPES 3. CANCER 4. ALS 5. HEPATITIS B 6. DIABETES 7. HUMAN
    PAPILLOMAVIRUS DISEASE(HPV) 8. ALZHEIMER 9. LUPUS (Lupus Vulgaris or Lupus Eratosthenes)ETC,,,,,, You can also visit his website:
    http://drchalaherbalhome.godaddysites.com
     email him on dr.chalaherbalhome@gmail.com

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  45. Hello everybody, i don't just know the reason why some people is finding it difficult to believe that there is a cure for herpes, i have been suffering from herpes since last three years but today i am happy that am cure from it with the herbal medicine of Dr Omongbe the great healer,i was browsing the internet searching for help when i came across a testimony shared by someone on how Dr Omongbe cure his herpes i was so much in need of getting his treatment but after all Dr Omongbe brought a smile to my face with his herbal medicine. I am so happy today that we have someone like this great healer out there, so my people out there kindly visit this great healer on his website https://dromongbenaturalre.wixsite.com/dromongbenaturalreme email address: dromongbenaturalremedies@gmail.com please sir keep your good work cause there are people out there who is in need of your healing medicine.once more contact him now you can call him or whatsApp his number +2348050407265

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  46. I thought my life had nothing to offer anymore because life became meaningless to me because I had HERPES virus, the symptoms became very severe and bold and made my family run from and abandoned me so they won’t get infected. I gave up everything, my hope, dreams,vision and job because the doctor told me there’s no cure. I consumed so many drugs but they never cured me but hid the symptoms inside me making it worse. I was doing some research online someday when I came across testimonies of some people of how Dr OLIHA cured them from HERPES virus, I never believed at first and thought it was a joke but later decided to contact him on the details provided and when I messaged him we talked and he sent me his herbal medicine and told me to go for a test after two weeks. Within 9 days of medication the symptoms disappeared and when I went for a test Lo and behold I was NEGATIVE by the Doctor Who tested me earlier. Thank you Dr OLIHA because I forever owe you my life and I’ll keep on telling the world about you. If you are going through same situation worry no more and contact Dr OLIHA on email   {oliha.miraclemedicine@gmail.com} or visit his website is https://olihamiraclemedicine.webs.com/     or  Cell Whatsapp Number +2349038382931 thank you  

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  47. To know more about Chief Dr Lucky you can visit his website (https://chiefdrluckyherbaltherapy.wordpress.com/)
     
     A friend that suffered from Herpes and was cure with the help of this great herbal doctor Chief Lucky so i decided to contact him for help in getting rid of my families genital herpes virus 1/2 which i did and all i was told to provide was just some useful information and some materials used in preparation of the natural cure and that i did and now i am the happiest person on earth because i am writing this testimony of mine with joy and happiness in my heart to the whole world to know that natural remedy is real because i am a living testimony of Chief Lucky traditional herbal cure and i want you all to contact Chief Lucky via his email: chiefdrlucky@gmail.com or WhatsApp him +2348132777335 am sure he will help you too.

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