Saturday, 23 May 2020

Hydractinia symbiolongicarpus and Hydractinia echinata: Cultivating a model colonial Hydrozoan Cnidarian in the lab.

Hydractinia symbiolongicarpus and Hydractinia echinata are sister species of colonial Hydrozoan Cnidarians. Hydractinia symbiolongicarpus occurs along the eastern coast of North America, from Maine to South Carolina. Hydractinia echinata found along North European Atlantic coasts. In the field, they are found exclusively on Gastropod Shells occupied by Hermit Crabs (e.g., Pagurus longicarpus). Colonies consist of polyps specialized for feeding, reproduction, or defense, which grow from a sheet of tissue called the stolonal mat. Unlike many of its Hydrozoan relatives, Hydractinia does not produce a free-living medusa stage (Jellyfish). Instead, gametes mature in a rudimentary medusoid that remains attached to sexual polyps. All polyps within a colony are clonally derived and therefore genetically identical. The mat consists of two epidermal cell layers, which sandwich a network of gastrodermal canals connecting polyps to each other and forming a gastrovascular system. Colonies grow by expanding the edge of the mat or by elongating individual stolons, extensions of gastrovascular canals encased in a thin, chitinous integument called the periderm. Colonies are dioecious and spawn about 90 min after first light. Eggs sink to the bottom after fertilization and develop into a planula larva within 2–3 days. Mature larvae latch onto a passing Hermit Crab shell by firing nematocysts located in their posterior ends. Once on the shell, the larvae metamorphose into a primary polyp in response to a Bacterial cue. The juvenile colony then grows as described, frequently covering the entire shell.

In a paper published in the journal EvoDevo on 26 March 2020, Uri Frank of the Centre for Chromosome Biology at the University of Ireland Galway, Matthew Nicotra of the Departments of Surgery and Immunology at the University of Pittsburgh, and Christine Schnitzler of the Whitney Laboratory for Marine Bioscience and Department of Biology at the University of Florida, discuss the cultivation of Hydractinia spp. in the laboratory, and its usefulness as a model organism.

Hydractinia morphology, life history, and culture. (a) Colony growing on a microscope slide. Major morphological structures are labeled. This colony was explanted from a larger colony. The yellow-brown rectangle at the center is a layer of chitin that is slowly deposited below the mat as the colony grows and indicates the outline of the original explant. Scale bar is 1 mm. (b) Life cycle of Hydractinia. (c) Typical setup of a 39 litre glass aquarium for culturing Hydractinia. Frank et al. (2020).

Cnidarians are an interesting and highly diverse group of Animals. This phylum diverged from the lineage leading to Bilaterian Animals (the group that includes Flies, Worms, and Vertebrates) at least 600 million years ago, providing sufficient time for substantial diversification within the Cnidarian lineage. Most extant Cnidarians share a body wall consisting of an epithelial bilayer, a gastric cavity, and a unique cell type, the stinging cell or cnidocyte (also known as nematocyte) from which the phylum name derives. Cnidarians are phylogenetically positioned as the sister group to Bilaterians; therefore, studying biological phenomena in Cnidarians can provide insight into their origin and how they have changed over evolutionary time between and within phyla. The past two decades have brought substantial progress in Cnidarian molecular biology and genetics, enabling functional genetic studies at least in some Cnidarian representatives. Overall, Cnidarians’ relative morphological simplicity, sequenced genomes, amenability to genetic manipulation, and phylogenetic position promise a fruitful future in research on these Animals that will inform areas spanning all the way from evolutionary biology to biomedical sciences.

Cladogram showing evolutionary relationships between Hydractinia and other model organisms. Frank et al. (2020).

Current research on Hydractinia focuses on a number of topics, including embryonic development, neurogenesis, stem cells, germ cells, and regeneration, allorecognition, metabolism, immunity, and natural product chemistry. Allorecognition refers to the ability to discriminate ‘self’ from ‘non-self’ within the same species, a phenomenon observed in most colonial Cnidarians, but not in Hydra or Nematostella, the two most commonly used Cnidarian model systems for molecular work. At present, Hydractinia is the only Cnidarian in which genes controlling allorecognition have been identified and functionally characterised. 

Other areas of interest are stem cells and regeneration. These topics have been well studied in Hydra and are emerging topics for Nematostella researchers too. Interestingly, data published to date suggest that both stem cell behavior and the mode of regeneration differ substantially between Cnidarian species. For example, Hydrozoan neuronal cells derive from migratory i-cells, whereas in Anthozoans, neural progenitor cells are epithelial. As to regeneration modes, Hydra can reform the main head structures following decapitation in the absence of cell proliferation whereas in Hydractinia and Nematostella cell proliferation is essential for regeneration. These findings highlight the importance of studying more than one animal in order to prevent false conceptual generalizations and underestimation of the complexity underlying biological phenomena.

Hydractinia does not show any evidence for age-related deterioration, s highly resistant to ionizing irradiation, and develops tumors only very rarely following genetic manipulation but not spontaneously. These features are consistent with high genomic stability in this Animal, a feature that remains to be investigated.

Manipulating gene expression has so far only been established in four Cnidarians: Hydra, Nematostella, Hydractinia, and Clytia. This can be done either by permanent modification of the Animal’s genome or by transient interference with specific gene products. Both approaches have their pros and cons and their usage depends on the type of experiment being conducted and availability of appropriate protocols for a given species and life stage.

The most common approach in Hydractinia is microinjection of nucleic acids and/or proteins into the zygote. Hydractinia spawning is light-induced without the need for any further induction. Eggs are not embedded in jelly and can be directly microinjected upon fertilisation. Electroporation techniques are currently being developed in Frank et al.’s labs with promising results. Circular plasmids readily integrate into the Hydractinia genome. The site of integration is unknown, but the process is highly efficient; in excess of 80% of injected embryos become transgenic in the hands of experienced researchers. This approach has been used to create fluorescent reporter lines for many developmental genes and cell type-specific markers. A more targeted way to genetically manipulate the Animals is provided by CRISPR–Cas9 technology (a technology that enables geneticists to edit parts of the genome, which was adapted from a naturally occurring genome editing system in Bacteria). In Hydractinia, this is performed by microinjecting site-specific short guide RNAs (sgRNA) together with recombinant Cas9 to generate loss-of-function mutations. Adding to the injecting cocktail a plasmid including a fragment of DNA, flanked by two homology arms, can be used for targeted knock-in of fragments. As with all plasmids, this DNA could also integrate randomly into the genome. Designing the injected DNA such that it must rely on the promotor of the target gene limits the likelihood that it would be expressed if integrating non-specifically.

Live imaging of transgenic Hydractinia gastrozooids. (a) A polyp expressing eGFP under an RFamide precursor promoter, labeling a subset of neurons. The animal was created via random integration of a circular DNA plasmid. (b) A polyp expressing eGFP under the endogenous Eef1a promoter. The animal was created using CRISPR/Cas9 to target integration of the eGFP coding sequence into the Hydractinia Eef1a locus. Frank et al. (2020).

Gene expression manipulation without genetic alteration can be achieved by injecting short hairpin RNA (shRNA) or morpholino oligonucleotides to lower expression of genes, or synthetic RNA to overexpress them. Finally, incubating polyps in seawater containing double stranded RNA (dsRNA) transiently lowers the expression of the corresponding gene, albeit with low efficiency. 

Hydractinia is also unique among model Cnidarians for being the only species in which a forward genetic approach has been used to identify the genetic basis of a phenotype. The reasons for this are almost entirely logistical. First, Hydractinia colonies can produce hundreds of embryos per day, making it possible to quickly generate large populations of bred Animals. Second, the Animals grow as encrustations on a surface that can be labeled, making it possible to co-culture large populations of genetically distinct animals in a small number of tanks. To date, forward genetic approaches have been used to identify genes responsible for allorecognition and sex determination. Given the availability of a sequenced genome and the cost efficiency of high-throughput genotyping, it seems feasible to consider mutagenesis screens as well.

An additional experimental approach in Hydractinia is grafting of tissues. This can be done for, e.g., introducing transgenic cells into a colony. Grafting of tissues from genetically distinct individuals requires at least partial matching of allorecognition alleles to prevent allogeneic rejection. 

Single-cell RNA sequencing methods are also under development in our labs with the first single-cell sequencing libraries giving encouraging results. Frank et al's current goal is to develop a robust cellular atlas to define major cell types and subtypes in Hydractinia and to identify marker genes for all cell types as was recently done in Hydra and Nematostella. With a robust genome and cellular atlas in place, Hydractinia will be poised to answer biological questions in a more comprehensive way. Flow cytometry and fluorescence activated cell sorting protocols are available, and together with many transgenic reporter strains it allows for generating cell type-specific transcriptomes following fluorescence activated cell sorting-sorting of defined cell populations.

As with any model organism, Hydractinia has limitations. Perhaps most obvious one is that it lacks a medusa stage, so researchers interested in this feature must look elsewhere, notably to the Hydroid Clytia and the Scyphozoan Aurelia. The existing Hydractinia research community also remains small compared to that for Hydra and Nematostella, so the availability of shared reagents and techniques is somewhat more limited. This concern is increasingly mitigated by additional labs beginning to study Hydractinia, and an upsurge in crosstalk between researchers.

The Hydractinia research community is relatively small but growing as Hydractinia is gaining recognition as a tractable Cnidarian research model. A recent National Science Foundation Enabling Discovery through GEnomic Tools grant has been awarded to Frank et al., ensuring that the genetic toolkit and community of Hydractinia researchers will continue to blossom and grow. Current resources include high-quality genomes and transcriptomes from both Hydractinia symbiolongicarpus and Hydractinia echinata. Draft Illumina genome and transcriptome assemblies are publicly available through the Hydractinia Genome Project Portal, and long-read PacBio genome assemblies for both species are forthcoming. With an estimated genome size of 774 Mb for Hydractinia echinata and 514 Mb for Hydractinia symbiolongicarpus, the Hydractinia genomes are larger than the genome of Nematostella (329 Mb) but smaller than that of Hydra (1086 Mb). Annotated reference genomes and transcriptomes can be used for mapping standard RNA sequencing data. Laboratory selected, fast-growing strains are available to anyone. Frank et al. are developing a community portal to be completed in the coming months, which will link to written and video-based protocols and to a community forum, and provide an online form to request Animals. Newcomers to the field are encouraged to attend the two biennial research conferences, the American Cnidofest and the European Tutzing meeting.

See also...

https://sciencythoughts.blogspot.com/2020/05/zhangiella-condensum-hydractinia.htmlhttps://sciencythoughts.blogspot.com/2019/10/portuguese-man-o-war-washing-up-on.html
https://sciencythoughts.blogspot.com/2019/10/velella-velella-hundreds-of-by-wind.htmlhttps://sciencythoughts.blogspot.com/2019/09/warning-issued-to-bathers-after-large.html
https://sciencythoughts.blogspot.com/2019/01/melicertissa-antrichardsoni-paralovenia.htmlhttps://sciencythoughts.blogspot.com/2019/01/physalia-physalis-thousands-stung-by.html
Follow Sciency Thoughts on Facebook.

23 comments:

  1. Hello my name is Kallya from USA i want to tell the world about the great and mighty spell caster called Priest Ade my husband was cheating on me and no longer committed to me and our kids when i asked him what the problem was he told me he has fell out of love for me and wanted a divorce i was so heart broken i cried all day and night but he left home i was looking for something online when i saw an article how the great and powerful Priest Ade have helped so many in similar situation like mine he email address was there so i sent him an email telling him about my problem he told me he shall return back to me within 24hrs i did everything he asked me to do the nest day to my greatest surprise my husband came back home and was crying and begging for me to forgive and accept him back he can also help you contact
    ancientspiritspellcast@yahoo.com or ancientspiritspellcast@gmail.com

    https://ancientspiritspellcast12.wordpress.com

    Blog: https://priestade.blogspot.com

    Website http://ancientspiritspellcast.website2.me

    WhatsApp +2347059715465 .






    Hello my name is Kallya from USA i want to tell the world about the great and mighty spell caster called Priest Ade my husband was cheating on me and no longer committed to me and our kids when i asked him what the problem was he told me he has fell out of love for me and wanted a divorce i was so heart broken i cried all day and night but he left home i was looking for something online when i saw an article how the great and powerful Priest Ade have helped so many in similar situation like mine he email address was there so i sent him an email telling him about my problem he told me he shall return back to me within 24hrs i did everything he asked me to do the nest day to my greatest surprise my husband came back home and was crying and begging for me to forgive and accept him back he can also help you contact
    ancientspiritspellcast@yahoo.com or ancientspiritspellcast@gmail.com

    https://ancientspiritspellcast12.wordpress.com

    Blog: https://priestade.blogspot.com

    Website http://ancientspiritspellcast.website2.me

    WhatsApp +2347059715465 .






    Hello my name is Kallya from USA i want to tell the world about the great and mighty spell caster called Priest Ade my husband was cheating on me and no longer committed to me and our kids when i asked him what the problem was he told me he has fell out of love for me and wanted a divorce i was so heart broken i cried all day and night but he left home i was looking for something online when i saw an article how the great and powerful Priest Ade have helped so many in similar situation like mine he email address was there so i sent him an email telling him about my problem he told me he shall return back to me within 24hrs i did everything he asked me to do the nest day to my greatest surprise my husband came back home and was crying and begging for me to forgive and accept him back he can also help you contact
    ancientspiritspellcast@yahoo.com or ancientspiritspellcast@gmail.com

    https://ancientspiritspellcast12.wordpress.com

    Blog: https://priestade.blogspot.com

    Website http://ancientspiritspellcast.website2.me

    WhatsApp +2347059715465 .

    ReplyDelete
    Replies
    1. My husband and I have been married for about 7 yrs now. We were happily married with two kids, a boy and a girl. 3 months ago, I started to notice some strange behavior from him and a few weeks later I found out that my husband is seeing someone. He started coming home late from work, he hardly care about me or the kids anymore, Sometimes he goes out and doesn't even come back home for about 2-3 days. I did all I could to rectify this problem but all to no avail. I became very worried and needed help. As I was browsing through the Internet one day, I came across a website that suggested that Dr Great can help solve marital problems, restore broken relationships and so on. So, I felt I should give him a try. I contacted him and he did a spell for me. Two days later, my husband came to me and apologized for the wrongs he did and promise never to do it again. Ever since then, everything has returned back to normal. I and my family are living together happily again.. All thanks to Dr Great . If you need a spell caster that can cast a spell that truly works, I suggest you contact him. He will not disappoint you. This is his E-mail: 

      infinitylovespell@gmail.com or infinitylovespell@yahoo.com

      WhatsApp +2348118829899

      Blog http://infinitylovespell1.blogspot.com








      My husband and I have been married for about 7 yrs now. We were happily married with two kids, a boy and a girl. 3 months ago, I started to notice some strange behavior from him and a few weeks later I found out that my husband is seeing someone. He started coming home late from work, he hardly care about me or the kids anymore, Sometimes he goes out and doesn't even come back home for about 2-3 days. I did all I could to rectify this problem but all to no avail. I became very worried and needed help. As I was browsing through the Internet one day, I came across a website that suggested that Dr Great can help solve marital problems, restore broken relationships and so on. So, I felt I should give him a try. I contacted him and he did a spell for me. Two days later, my husband came to me and apologized for the wrongs he did and promise never to do it again. Ever since then, everything has returned back to normal. I and my family are living together happily again.. All thanks to Dr Great . If you need a spell caster that can cast a spell that truly works, I suggest you contact him. He will not disappoint you. This is his E-mail: 

      infinitylovespell@gmail.com or infinitylovespell@yahoo.com

      WhatsApp +2348118829899

      Blog http://infinitylovespell1.blogspot.com









      My husband and I have been married for about 7 yrs now. We were happily married with two kids, a boy and a girl. 3 months ago, I started to notice some strange behavior from him and a few weeks later I found out that my husband is seeing someone. He started coming home late from work, he hardly care about me or the kids anymore, Sometimes he goes out and doesn't even come back home for about 2-3 days. I did all I could to rectify this problem but all to no avail. I became very worried and needed help. As I was browsing through the Internet one day, I came across a website that suggested that Dr Great can help solve marital problems, restore broken relationships and so on. So, I felt I should give him a try. I contacted him and he did a spell for me. Two days later, my husband came to me and apologized for the wrongs he did and promise never to do it again. Ever since then, everything has returned back to normal. I and my family are living together happily again.. All thanks to Dr Great . If you need a spell caster that can cast a spell that truly works, I suggest you contact him. He will not disappoint you. This is his E-mail: 

      infinitylovespell@gmail.com or infinitylovespell@yahoo.com

      WhatsApp +2348118829899

      Blog http://infinitylovespell1.blogspot.com

      Delete
    2. Hello everyone i Am williams pater and i am from USA i am here to give my testimony about an herbal doctor called Dr,olu I was heartbroken because i had very small penis,not nice to satisfy a woman, i have been in so many relationship, but cut off because of my situation, i have used so many product which doctors prescribe for me, but could not offer me the help i searched for. i saw some few comments on the internet about this specialist called Dr,OLU and decided to email him on his email i saw on the internet,(drolusolutionhome@gmail.com ) so I decided to give his herbal product a try. i emailed him and he got back to me, he gave me some comforting words with his herbal product for Penis Enlargement, Within three weeks of me use it, i began to feel the enlargement, " and now it just 4 weeks of using his products my penis is about 8 inches longer, and i had to settle thing out with my ex girlfriend , i was surprised when she said that she is satisfied with my performance in bed and i now have a large penis.thanks to DR OLU for is herbal product. you can also reach him with emsil  drolusolutionhome@gmail.com though is..number WHATASPP him today on this number [ +2348140654426 ] 

      Delete
    3. Am so happy to share this testimony with everyone about how i got my herpes cured from Dr. Obela the godfather of herbal medicine. I had been going from one country to another for medication for 3 years just to be cured from herpes but all seems in vain, i was confused and decided to search through the Internet if i could get any news concerning herpes so i do keep searching until i visited one particular site and i saw the great testimonies from different persons testifying how Dr. Obela has cure them from herpes and other diseases and i was shock and was thinking if this is real, after a due consideration i decided to contact him with the email address I found in the posted just to give it a trial and when i contacted him he proved to me that his herbal medicine can cure me if only i believe in him, so i requested for his herbal medicine and i got it and i applied it as prescribed and after 1weeks and 3 days of application i went for a test and to my greatest shock the herpes virus has disappeared and it was just like a dream. Today I am cured from herpes am so happy and thanking God for using Dr. Obela to cure me. so if you are out there and you are suffering for herpes cure or any kind of illness kindly contact Dr. Obela through he will help you to cure that illness. you can get in touch with him via: drobelaspellhome@gmail.com or call/whatsapp him on this number +2347084171950.. Thanks very much DR. Obela
      he can also cure all type of disease like:
      HIV/AID .... cancer disease......e.t.c

      Delete
    4. i want to testify of how DR MOSES  herbal mixture cream enlarged my penis size. i never thought i will be happy again in my life because of my small penis situation,i have struggling with this issue my life time and i could not find a professional result i wanted ,i ave went to hospitals,used vaccines,tried several herbs online ,penis pumps and all but none rendered me the solution i wanted ,i felt like killing when my fiance KIMBERLY left me after undressing myself in front of her,my size was too embarrassing and shameful imagine a 37 years old man like me having 3.5 inches penis size..the world seem to an end to me when she left,  one faithful day as i was watching a video on you tube i saw on the comments of one MR PAUL HAVERSACK testifying of this great herbal healer DR MOSES helped him enlarge his penis .i was shocked and happy so i quickly visited his website and emailed him within 30 mins he got back to me and told me all i need to buy and i did so after 4 days i received his herbal medicine ,he gave me instructions on how to use it ,as i am speaking to you people now after using the cream for just two weeks my penis size is 10 inches long and 8.0 girth ,,am so happy and grateful for his work in my life thank you so much Dr MOSES ,,i also learnt he has cure for LOW SPERM COUNT,PREMATURE EJACULATION,ERECTILE DYSFUNCTION,HIV/AIDS VIRUS,DIABETES 1/2,HERPES DISEASE,CANCER,and lots more you can email him on ( buba.herbalmiraclemedicine@gmail.com ) or call/whats-app him directly on +2349060529305 thanks or you can call me 05019914802 am from Royal Arkansas united state or You can go through is website https://bubaherbalmiraclemedicine.wordpress.com/

      Delete
  2. Hello everyone, Are you looking for a professional trader, forex and binary manager who will help you trade and manager your account with a good and massive amount of profit in return. you can contact MR. GAVIN RAY for your investment plan inorder to get started. for he helped me earned 11,300usd with a little investment money. Gavin ray you're the best trader I can recommend for anyone who wants to invest and trade with a genuine trader, he also helps in recovery of loss funds..you can contact him on his Email: gavinray78@gmail.com WhatsApp: +1 (217) 963-5382

    ReplyDelete
  3. i visited different hospital but they gave me list of expensive drugs to treat the symptoms and never cured me. A closed friend of mine introduce me to a Herbal Doctor who cured her. I was scared to contact him because of what i experienced from fake doctors. she encouraged me and gave me his whatsapp contact,and i contacted him and i asked him series of questions, i also ask him to show me the results of those he have cured with his herbal drugs,and he gave me all the information i needed that gave me hope. He send a Herbal medicine to me that i took and it seriously worked for me. am now Hepatitis negative. God bless you for being a sincere and great man. Am so excited and am free from herpes virus. you can reach him via email;dralikaherbalhome@gmail.com or whats-app/call on +2348116570358

    ReplyDelete
  4. i visited different hospital but they gave me list of expensive drugs to treat the symptoms and never cured me. A closed friend of mine introduce me to a Herbal Doctor who cured her. I was scared to contact him because of what i experienced from fake doctors. she encouraged me and gave me his whatsapp contact,and i contacted him and i asked him series of questions, i also ask him to show me the results of those he have cured with his herbal drugs,and he gave me all the information i needed that gave me hope. He send a Herbal medicine to me that i took and it seriously worked for me. am now Hepatitis negative. God bless you for being a sincere and great man. Am so excited and am free from herpes virus. you can reach him via email;dralikaherbalhome@gmail.com or whats-app/call on +2348116570358

    ReplyDelete
  5. QUICK RELIABLE CURE, Am so glad expressing my profound gratitude to a man that is so concerned about other people's well being. I was suffering from (HERPES SIMPLEX VIRUS) over 4 years, i have spent thousands of dollars in buying expensive drugs and visiting several hospitals in different countries, i was really in severe pain "until" one glorious morning, i read about Doctor Razor on the internet on how he had helped and cured so many people suffering from HERPES 1&2, HPV, CANCER, HIV/AIDS, ALZHEIMER, FIBROID, ENLARGED PROSTATE, HEMORRHOIDS' PILE and HEART DISEASE ETC. So i contacted him through this Email: drrazorherbalhome@gmail.com and i shared my problems with him, he gave me the healing procedure and i followed them. 18 days later he told me to go for a medical test, behold the result was negative, now am (HERPES SIMPLEX VIRUS)  Negative. I just want to use this platform to thank Doctor Razor for curing me from that deadly virus and also for exposing his herbal cure to the world, please you can as well contact him at drrazorherbalhome@gmail.com (or) Whatsapp/Call him +2349065420442 for solutions to your health problems

    ReplyDelete
  6. QUICK RELIABLE CURE, Am so glad expressing my profound gratitude to a man that is so concerned about other people's well being. I was suffering from (HERPES SIMPLEX VIRUS) over 4 years, i have spent thousands of dollars in buying expensive drugs and visiting several hospitals in different countries, i was really in severe pain "until" one glorious morning, i read about Doctor Razor on the internet on how he had helped and cured so many people suffering from HERPES 1&2, HPV, CANCER, HIV/AIDS, ALZHEIMER, FIBROID, ENLARGED PROSTATE, HEMORRHOIDS' PILE and HEART DISEASE ETC. So i contacted him through this Email: drrazorherbalhome@gmail.com and i shared my problems with him, he gave me the healing procedure and i followed them. 18 days later he told me to go for a medical test, behold the result was negative, now am (HERPES SIMPLEX VIRUS)  Negative. I just want to use this platform to thank Doctor Razor for curing me from that deadly virus and also for exposing his herbal cure to the world, please you can as well contact him at drrazorherbalhome@gmail.com (or) Whatsapp/Call him +2349065420442 for solutions to your health problems

    ReplyDelete
  7. Hello everyone i am very delighted that i recently got cured of herpes virus. I have been battling with this disease for over 4 years now, ever since i got diagnosed of the virus i have been using medical pills and therapies to suppress the virus. But inside of me i needed the Virus gone cause my health was deteriorating, So  i went on to the internet to search on how one could get cured from this virus, when i came across some testimonies on how herbalist razor has helped them get rid of Herpes virus, Hiv/aids, Cancer. I copied out his details (email ; drrazorherbalhome@gmail.com) and Whatsapp number +2349065420442) as provided by the testifiers on a blog site. But before that i was so skeptical, i knew there are so many herbal doctors on the internet,i needed the one who can help me get through this Herpes Virus completely with his herbal Remedy. I got in contact with doctor Razor,and i explained my problem to him and he promised to help me, which he actually did. I got my medication in the US sent by doctor razor and he instructed me on how i was to take it for 18 days. After the said 18 days i went for medical check up and the doctors ran some test on me and told me i was Herpes Negative, i was amazed and dumb-folded, so i had to go back home, i noticed my health for some months, i didn't have any outbreaks nor symptoms of the virus, but i went to another hospital after 2 months to conduct another test and the same result. I will be forever grateful to you doctor Razor and I will make sure the world gets to know about your herbal exploits sir. Reach doctor razor on his email : drrazorherbalhome@gmail.com or whatsapp/call his cell on +2349065420442

    ReplyDelete
  8. I am so happy that i got cured from herpes virus 1&2 after i have suffered from this virus for over 3 years. I have been using drugs to suppress it,i got introduced to doc tor razor by a friend of mine who knew i was herpes positive, she believes in herbs, and i have full doubt on her idea to try out herbal medicine. I contacted the doctor and told him what i was passing through, so he promised he was going to get me cured completely. I am so glad that after applying doctor razor herbal remedy for 18 days there is no trace of the virus in my body as confirmed by the results given to me by the medical hospital,where i conducted my test. I am so grateful Doctor Razor. For those who are in same situation just like i was, you can reach out to doctor razor on email : drrazorherbalhome@gmail.com , WhatsApp or Call him on +2349065420442. Please share as received, so multitude can be saved from this virus.

    ReplyDelete
  9. I am so happy that i got cured from herpes virus 1&2 after i have suffered from this virus for over 3 years. I have been using drugs to suppress it,i got introduced to doc tor razor by a friend of mine who knew i was herpes positive, she believes in herbs, and i have full doubt on her idea to try out herbal medicine. I contacted the doctor and told him what i was passing through, so he promised he was going to get me cured completely. I am so glad that after applying doctor razor herbal remedy for 18 days there is no trace of the virus in my body as confirmed by the results given to me by the medical hospital,where i conducted my test. I am so grateful Doctor Razor. For those who are in same situation just like i was, you can reach out to doctor razor on email : drrazorherbalhome@gmail.com , WhatsApp or Call him on +2349065420442. Please share as received, so multitude can be saved from this virus.

    ReplyDelete
  10. Am so happy to share this testimony with everyone about how i got my herpes cured from Dr. Obela the godfather of herbal medicine. I had been going from one country to another for medication for 3 years just to be cured from herpes but all seems in vain, i was confused and decided to search through the Internet if i could get any news concerning herpes so i do keep searching until i visited one particular site and i saw the great testimonies from different persons testifying how Dr. Obela has cure them from herpes and other diseases and i was shock and was thinking if this is real, after a due consideration i decided to contact him with the email address I found in the posted just to give it a trial and when i contacted him he proved to me that his herbal medicine can cure me if only i believe in him, so i requested for his herbal medicine and i got it and i applied it as prescribed and after 1weeks and 3 days of application i went for a test and to my greatest shock the herpes virus has disappeared and it was just like a dream. Today I am cured from herpes am so happy and thanking God for using Dr. Obela to cure me. so if you are out there and you are suffering for herpes cure or any kind of illness kindly contact Dr. Obela through he will help you to cure that illness. you can get in touch with him via: drobelaspellhome@gmail.com or call/whatsapp him on this number +2347084171950.. Thanks very much DR. Obela
    he can also cure all type of disease like:
    HIV/AID .... cancer disease......e.t.c

    ReplyDelete
  11. 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
  12. i visited different hospital but they gave me list of expensive drugs to treat the symptoms and never cured me. A closed friend of mine introduce me to a Herbal Doctor who cured her. I was scared to contact him because of what i experienced from fake doctors. she encouraged me and gave me his whatsapp contact,and i contacted him and i asked him series of questions, i also ask him to show me the results of those he have cured with his herbal drugs,and he gave me all the information i needed that gave me hope. He send a Herbal medicine to me that i took and it seriously worked for me. am now Hepatitis negative. God bless you for being a sincere and great man. Am so excited and am free from herpes virus. you can reach him via email;dralikaherbalhome@gmail.com or whats-app/call on +2348116570358.

    ReplyDelete
  13. I have been battling this Herpes disease for almost 1year now....I tried all possible means to get cure from my Herpes Disease but all to me in vain until i saw a post in a health forum about a herbal doctor  Dr Alika 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...then i contact Dr Alika and he replied me and explain how the process work so after ordering for the medicine I got it within 3/4 working days through UPS Delivery Service and I took it according to the way Dr Alika instructed, I was so happy after 14days I took the medicine there was a very big changes 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 Alika to get ride of these Herpes Virus forever contact him via email on dralikaherbalhome@gmail.com or whatsapp/call him on  +2348116570358.

    ReplyDelete
  14. I Want To Appreciate Dr.OYAGU for his great deeds, I Was Diagnosed With type 2 Herpes Virus Last year,And i Was Looking For Solution To Be Cured Luckily I Saw Testimonies On How Dr.OYAGU Cure Herpes Virus I Decided To Contact Dr.OYAGU I Contacted Him He Prepared A Herbal Medicine Portion And Sent It To Me,I Started The Herbal Medicine For My Health.He Gave Me Step By Step Instructions On How To Apply It, When I Applied It As Instructed, I Was Cured Of This Deadly Herpes Within 2 weeks, I Am Now Herpes Negative.My Brother And Sister I No That There Are So Many People That Have The Same Herpes Virus Please contact Dr OYAGU To Help You Too,And Help Me To Thank Dr.OYAGU For Cure Me, I’m Cured By Dr. OYAGU Herbal Medicine,His Contact Email:oyaguherbalhome@gmail.com

    Or Cell Whatsapp Number +2348101755322 thank you

    ReplyDelete
  15. I Want To Appreciate Dr.OYAGU for his great deeds, I Was Diagnosed With type 2 Herpes Virus Last year,And i Was Looking For Solution To Be Cured Luckily I Saw Testimonies On How Dr.OYAGU Cure Herpes Virus I Decided To Contact Dr.OYAGU I Contacted Him He Prepared A Herbal Medicine Portion And Sent It To Me,I Started The Herbal Medicine For My Health.He Gave Me Step By Step Instructions On How To Apply It, When I Applied It As Instructed, I Was Cured Of This Deadly Herpes Within 2 weeks, I Am Now Herpes Negative.My Brother And Sister I No That There Are So Many People That Have The Same Herpes Virus Please contact Dr OYAGU To Help You Too,And Help Me To Thank Dr.OYAGU For Cure Me, I’m Cured By Dr. OYAGU Herbal Medicine,His Contact Email:oyaguherbalhome@gmail.com

    Or Cell Whatsapp Number +2348101755322 thank you

    ReplyDelete
  16. I Want To Appreciate Dr.OYAGU for his great deeds, I Was Diagnosed With type 2 Herpes Virus Last year,And i Was Looking For Solution To Be Cured Luckily I Saw Testimonies On How Dr.OYAGU Cure Herpes Virus I Decided To Contact Dr.OYAGU I Contacted Him He Prepared A Herbal Medicine Portion And Sent It To Me,I Started The Herbal Medicine For My Health.He Gave Me Step By Step Instructions On How To Apply It, When I Applied It As Instructed, I Was Cured Of This Deadly Herpes Within 2 weeks, I Am Now Herpes Negative.My Brother And Sister I No That There Are So Many People That Have The Same Herpes Virus Please contact Dr OYAGU To Help You Too,And Help Me To Thank Dr.OYAGU For Cure Me, I’m Cured By Dr. OYAGU Herbal Medicine,His Contact Email:oyaguherbalhome@gmail.com

    Or Cell Whatsapp Number +2348101755322 thank you

    ReplyDelete
  17. I am Monisha Alexis, I promise to share this testimony all over the world once my boyfriend returns back to me, and today with all due respect I want to thank DR.ONIHA for bringing joy and happiness to my relationship and my family. I want to inform you all that there is a spell caster that is real and genuine. I never believed in any of these things until I lost my boyfriend, I required help until I found a great spell caster, And he cast a love spell for me, and he assured me that I will get my boyfriend back in two days after the spell has been cast. Three days later, my phone rang, and so shockingly, it was my boyfriend who has not called me for the past 6 years now, and made an apology for the heart break, and told me that he is ready to be my backbone till the rest of his life with me. DR.ONIHA released him to know how much I loved and wanted him. And opened his eyes to picture how much we have shared together. As I`m writing this testimony right now I`m the happiest girl on earth and me and my boyfriend are living a happy life and our love is now stronger than how it was even before our break up. So that`s why I promised to share my testimony all over the universe. All thanks goes to DR.ONIHA for the excessive work that he has done for me. Below is the email address in any situation you are undergoing, it may be a heart break, and I assure you that as he has done mine for me, he will definitely help you too. 
    EMAIL: ONIHASPELLTEMPLE@GMAIL.COM
    WEBSITE: http://onihaspelltemple.com 
    CALL/WHATSAPP : +16692213962. 

    ReplyDelete
  18. My Name is Dr abaka you can Contact Me via Email drabakatemple@gmail.com For Penis Enlargement Product to help you get as long as 8inches Long with good Erection. Contact Me Via Email : drabakatemple@gmail.com   Via WhatsApp +2347010538590 

    ReplyDelete
  19. God bless Dr. USELU for his marvelous work in my life, I was diagnosed of HERPES SIMPLEX VIRUS since 2018 and I was taking my medications, I wasn’t satisfied i needed to get the HERPES out of my system, I searched about some possible cure for HERPES i saw a comment about Dr. USELU , how he cured HERPES 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 within 3 days. I took the medicine as prescribed by him and 2weeks later i was cured from HERPES contact him via email (dr.uselucaregiver@gmail.com) once again thanks to you Dr. USELU cure the flowing virus, contact his email or add him on whatsapp (+2348132015009) cancer cure
    diabetes cure
    ringing ear
    herpes cure
    warts cure
    HPV cure
    HIV cure
    get your ex back
    pregnancy herbal medicine
    Hepatitis  

    ReplyDelete