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The Corroboree


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Posts posted by Darklight

  1. Ha! Found em.


    They'd sent some NTS 4/20 and some NTS Bio-N. I know why they labelled it the way they did, because proprietary blends can change over time.


    Everything triangulates nicely via notes and cached web pages and backup drives. So yeah, lab notes still winning :D

  2. About a gazillion years ago  ( OKOK it was 2013 ) a lovely kind forum member sent me some microbial fertiliser product samples. I have the envelope but the address is faded and lost to time and fridge mould, but it was a NSW postcode.


    From memory there were 2 aliquot tubes in it. I still have one, labelled Azotobacter.


    The other tube is missing, maybe used up. I can't remember what it was labelled as.


    Was about to start some plant rooting experiments for some putative mutants which are proving a right bastard to deflask. Looking back at the 2013 notes they say that the control species was successfully rooted in-vitro using Nutritech NTS 4/20. The deflasks survived well and went on to propagate so well we abandoned micropropagation altogether


    Nutritech NTS 4/20 is a bacterial product. It's been discontinued, but it did contain Azotobacter.


    There's a chance that the lost test tube contained the proprietary blend NTS 4/20 and the Azotobacter is a pure product from another source


    The other option is that the tube labelled Azotobacter is NTS 4/20 and the lost tube was something else


    Fucked if I can find the original PM via search engine. And I'm about to load up the autoclave.


    If it was you, you lovely person, do you remember what was in the other tube?


    And do I owe you anything for the trade? Did I send you anything?


    I lost a bunch of references to the trade during an OS crash this month- my scratchpad didn't back up as well as I was hoping.


    This is entirely the reason I keep lab notes. But even these weren't sufficient in this instance.

  3. 3 minutes ago, MeanGreen said:

    Especially when you consider that big "false kratom" debacle back in the early 2000's where my compatriots Bruno & friends at ebotashop flooded the market with false kratom (M. parvifolia I believe). This could have caused everyone to just brush off kratom as ineffective and killed the market in the egg...


    There were vendor pages online back then which incorrectly ( and possibly knowingly ) pushed the psychedelic properties of M. speciosa and those most definitely contributed to the TGA scheduling. Mitragyna speciosa is not psychedelic, had never been touted as psychedelic prior to the existence of those few web pages, and the incorrect publicity was not scientifically evaluated, but was used directly as evidence nonetheless during the TGA scheduling


    By the time Jon Hanna's expose on false kratom came out it was too late to do anything


    Flawed data met lack of oversight and flawed process and produced poor outcomes



    3 minutes ago, MeanGreen said:


    I think it's extremely unfortunate that Australia decided to ban it, especially during an opiate epidemic. How can you consider an opioid which doesn't cause lethal respiratory depression as not having any medicinal potential is beyond me...


    It also has potential as an anti-inflammatory, anti-tumour etc. It was a cruel and irrational scheduling, and because of the bureaucratic specifics of the scheduling process back then we were not permitted to respond or object to the scheduling. I believe that's changed for all new introductions, T would know better than I, but it's not retrospective


    If we'd not been hamstrung by the scheduling we were on track to do so much more good research


    In practical terms a helluva lot of good science gets proven wrong over time ( which is entirely the point ) and so many claims made back then could have been fortified by extra research or dropped as spurious


    Our facility, our towns and region really lost out on this. Research programs and commercial opportunities all happened off shore. Quite a few overseas companies have done seriously well out of it and should continue to do so for some time. Good on 'em. We were uniquely placed to do all that here, in a region of high biodiversity, appropriate climate, high unemployment and fair access to some first rate research facilities. But hey, no.


    I still want that small, private research lab where we can rigorously train citizen scientists in the love and practice of quality empirical data generation and never be de-funded by any bastard government again. I want it so bad I can taste it. So much talent going to waste.



    3 minutes ago, MeanGreen said:


    Could you share a bit about what the selection process was for the Rifat clone? Was it simply the most vigorous sprout of the batch of seeds?


    Erk. It was the only one which survived. We bit our nails for a year or so until it was proven both stable in micropropagation and active. Long story snipped


    3 minutes ago, MeanGreen said:


    If you do a talk about this at EGA I might just have to fly my ass over there haha :)


    Seriously, you should come to an EGA anyhow. They're deadset brilliant. Best crew ever. And if we are accepted for an EGA presentation on this and T agrees to co-present with me again I may hold you to that statement :D

    • Like 6

  4. On 1/11/2018 at 7:45 AM, sagiXsagi said:

    Retrospective notes #2 : drunk but wise 


    you might find that I am less into conclusions and more into random notes in this 1.5 year report, but hey, I hope I can arrange them notes better, in this single post and try to give it a structure.



    Mate, it's a hard task and a big ask. But it will be really worth doing as a single document. It'll prolly also help formalise a lot of the notes into new ideas. And when it's finished it will be gorgeous :)


    I write lab notes like this, all good :)

    • Like 1

  5. I have a Griffonia simplicifolia growing in a pot at home, and these popped up last summer. Until last week I thought they were new shoots from the vine and was waiting for them to develop leaves. After 15 months waiting I had a closer look last week and they look like bloody Ephedra!


    Not sure which one. I recycle my propagation mix and throw a lot of old prop mix into different pots to top them up. They could be *really* old E. gerardiana from 20 years ago, E. major or E. sinica from seed that didn't germinate over 5 years ago.


    Whoever they are they put a huge smile on my face. Never give up :)


    Thanks for keeping us updated, this is great! Sorry I didn't get to mention this thread at T's and my EGA presentation on research. It was in my notes to do but I got nervous and left heaps of major points out :/


    Hope I'm not interrupting here, lemme know and I'll move it



    • Like 1

  6. Yup- sleep and constipation are huge barriers in PD


    Melatonin is excellent for sleep for a lot of ppl


    Melatonin ref from 2011


    I know the kefir sounds like too simple a constipation solution to be effective, but please let me know how it goes for you. It's such an easy place to start and if you can't make your own, local Farmers Markets often sell them, or find a good quality local fermented foods person on FB. Is a bit of a thing right now so locating someone with experience could be easy

    • Like 1

  7. On 1/5/2018 at 10:17 PM, paulinepie said:

    I have researched the medicinal cannabis, but currently Parkinsons is not on the included allowable program. Apparently the best is getting the fresh bud and juicing it immediately. However, we are not growing it due to regulations and our urban property will not sustain the space required.


    Now that is a shame :( Are there any informal dispensaries in Perth you know of?


    I would def acquire a script for melatonin for your husband, it's becoming increasingly recognised as beneficial for Parkinsons. Unlikely to interact AFAIK but please do your own checking of anything I say


    Wrt Lions Mane I'd advise caution and small doses. In fact given the Sifrol/ Mucuna etc etc I'd advise against it. I read something about it potentiating psychosis in some late stage Parkinsons patients. I lost those refs a while back and am chasing them up again.


    There are a bunch of variables around that statement above which do need checking


    Have you used it earlier in your husband's PD progression?


    With you in hope

  8. Further, given the gut bacteria changes implicated in Parkinsons progression, have you tried either water kefir or milk kefir?


    In addition to slowing gut bacterial changes it can improve gut motility and decrease constipation in healthy individuals. There is potential for it's regular consumption in Parkinsons patients but I have yet to see any academic data on it.


    As gut bacteria studies are a relatively new field the absence of formal data doesn't surprise me too much. Because it is unlikely to interfere with pharmaceutical drug actions I'd be giving it a go for a week or so. This, however is just my untested opinion


    My relative with longer term Parkinsons will not try it. My relative with the rapid-progression form cannot tolerate water kefir at all- we have yet to try them on milk kefir


    We make our own water kefir from fresh grains. It all sounded terribly hippy-ish to start with back in the day, but there are a few of the fam who are deriving benefit from it and the taste is pretty good so it's a winner

    • Like 3

  9. On 12/22/2017 at 7:36 PM, paulinepie said:

    I am treating my husband's Parkinson's Disease Symptoms with a few doses of Velvet bean. It is as effective as taking the pharmaceuticals - Sinamet, which contains both levodopa (the good guy) and Carpidopa (seriously bad for humans).

    This and Ashwaganda are effective in controlling the symptoms with no apparent side effects.


    Has anyone else used VB in this way or used any other alternatives for Parkinsons.


    Hi Paulinepie, so sorry you both have to work with Parkinsons and thank you so much for the info. I find it really valuable to hear about direct experience with treating Parkinsons with natural products


    May I ask what stage your husband's Parkinsons is at, and how long he has been taking Mucuna ( Velvet bean ) for?


    Hadn't heard of using Withania ( Aswagandra ) for Parkinsons but I have used it as a general health product previously and found it really well tolerated and effective- thank you again


    A few of my close relatives have Parkinsons- one with the longer term form ( +20 years since diagnosis, now late stage + dementia ) and two with the faster term form ( one has passed, the other diagnosed recently and progressing rapidly )


    My family member with the longer term form is remarkably resistant to trying anything not prescribed by the GP or specialist, which is frustrating but is their choice.


    When I saw them last they were willing to try medical cannabis ( informally sourced ) high CBD tincture ( unsupported by formal analysis but a widely distributed form ) to combat the frozen state and hopefully reduce tremors.


    However his tremors and frozen state are quite advanced, and the tincture wasn't suitable to titrate a dose with in my absence- it was too slippery to measure small amounts and would require some dilution. His partner was not at all supportive of the treatment and informed me that they would not continue with it in my absence as his tremors would not allow him to dose alone reliably. We ended up not trialling the product at all.


    I've heard high CBD products can be effective for Parkinsons tremors, freezing etc and if you are comfortable trialling and able to source some I'd recommend beginning with a low dose and set a comfortable amount as soon as possible after diagnosis.


    If your clinician is comfortable with CBD co-administration with pharmaceuticals I'd confer with them. My relative is off all anti-Parkinsonians at this late stage but is taking antipsychotics for the delusions- they have been helpful, but potential contraindications with the antipsychotics were another reason for not undertaking the CBD even at small doses and they do not wish to confer on this matter with their clinician.


    Thank you so much for your valuable insights into potential non-pharma treatments. I wish you both, and your family, all the best on this difficult journey. If you are able to share further experience I'd love to hear of it, either online here or via PM



    • Like 4

  10. O M G thank you EGA crew for another excellent weekend. So much fun and so very professional all at once <3


    Righto, I didn't see many actual presentations cos I was largely freaking out trying to finesse my own ( shared ) upcoming workshops, but that's OK as a bunch of them are coming up on the EGA YouTube channel


    Any time I was outside I'd walk ten paces and run into someone I hadn't seen in five years and we'd get talking


    Which is perfect for me as that's exactly what I wanted to do. Personally EGA is, for me, about networking and catching up and trying to pitch in at that level of excellence that the other EGA crew show


    Also I got to hang out with Kaz at the EGA info tent which is always a goal for me


    But there's always something for everyone at an EGA, and everyone's EGA is accordingly different.


    Special thanks to Obtuse for sharing the cloning workshop facilitation ( halfa beta blocker rocked for me and was the first time I'd ever felt comfortable presenting anything ), mate you rocked and I would love to co-present another if you'd have me. The people who came along were wonderful and involved and we kinda went overtime but thankfully only into lunch break


    Special thanks to Torsten for saving my stuttery arse at our Experiment Design workshop when I was nervous even tho we had rehearsed it so many times. Same on the co-presentation, next time I'll do it better and actually get all the crucial points across


    Melbs crew, EGA crew, you rock. I had the best time. Your professionalism, compassion, humour, comprehensible cohesiveness and sense of fun... whoa... What a great weekend.


    Also: while in transit I musta eaten halfa VIC. The entire place is delicious. Wagon Wheels the size of yer head at Yarck, off-the-track pubs providing exquisite burgers for $$fuckall. Even the round trip is fun


    So: how was your EGA?

    • Like 3

  11. 21 hours ago, shonman said:

    Thanks Darklight!


    No wuckas, is a pleasure walking with you on the path, your thorough note taking, good questions and sharing results here is an inspiration


    21 hours ago, shonman said:

    Yes, always doing 1/2 strength, 1/2 sugar to start out.


    :) Sorry i shoulda checked yr earlier responses


    21 hours ago, shonman said:

    i agree, PPM suppresses, but does not kill some or many? Fungi.

    sometimes it is very evident, in that if I leave plants in culture too long

    at a certain stage the PPM wears off or is no longer effective.


    This is very true. It also has a use by date. I know this because I used some really old stock once past the date and it wasn't effective. Lesson learned



    21 hours ago, shonman said:

    i think some species of fungi are possibly symbiotic with the plants in some cases

    but in vitro, or in humid conditions with no ventilation....the fungus takes over.


    This is also true IME


    21 hours ago, shonman said:

    or maybe it's just contamination.


    It often is, but not always. Depends on circumstance and your experience with the workflow for whichever species it is


    21 hours ago, shonman said:

    Re: cabrerana....I will probably use ppm at first, just in hopes of getting something to work.

    then make sure those cultures get changed, then deflasking, before ppm wears off.

    then, I will let the ppm wear off or discontinue in some test cultures, and see how those do.

    if they have contaminants still, 

    i might t......( snipped this bit )


    Ye gods it all sounds arduous, I feel your pain. Scattergun approach is probably best at this point and at least you're taking good notes and making sure you have backup material.


    It's hard starting cultures for a species new to TC when you have very little parent material, and heartbreaking. Worth it when it works tho



    21 hours ago, shonman said:

    right now I am 'afraid' to take any growth from the plant.


    No rush, let it grow back and be strong


    21 hours ago, shonman said:

    have been so busy....when I can I will slow down and really tune into the plants energy...

    'ask it, what it wants/ needs to grow '

    the other day when I was tending to them, I felt a tingle when the healthiest one brushed against my arm.

    almost a sort of loving, grateful energy like ' thanks for taking care of us '

    I will take some time to really get what this plant needs, from a centered space.



    That's completely unscientific of you and is exactly what I do all the time too :)



    20 hours ago, ☽Ţ ҉ĥϋηϠ₡яღ☯ॐ€ðяئॐ♡Pϟiℓℴϟℴ said:


     Darklight, are you the one we can thank for being able to grow our fave Acacia? (phleb)



    Nup, not down to me at all. I had a few years of having a good crack at aseptic culture, as did a well known TC local who had similar luck. Another young fella took the population pressure off the phlebs by identifying other easy to cultivate Australian acacias with a similar profile- so there was no longer a need to hunt the phlebs specifically in the wild. Then a third young fella found a rhizobium inoculant which promotes survival past the critical time point where most of the people cultivating it were experiencing progressive dieback which was ultimately fatal.


    Point being this project is a community effort. Not deliberately, but it didn't happen in isolation. And some outcomes ( like the reduced population pressure from wild harvesting ) were unanticipated and secondary to other research. And a good many people posted and shared results both formally and informally which allowed patterns to emerge and be followed. Lots of things didn't work- until a few did.


    I'm not 100% confident the species is out of the woods yet ( har har ) but it's looking to be in better shape wrt numbers and cultivation than it was 20 years ago. Keep an eye on it, research where you can, and understand that collective direct community input has the capacity to trump government grant applications for actually making things happen

    • Like 1

  12. Quote

    6x Aseptic cultures of an Acacia (for tissue culture) incl 100ml tubes & 30ml media – Priceless – Darklight


    Just confirming- those aseptic Acacia in tissue culture are Acacia obtusifolia.


    Dunno about priceless, they're as valuable as you make them :) Practice your deflasking, run simple tissue culture experiments on 'em, mutate the hell out of em, do what you like

    • Like 1

  13. On 11/13/2017 at 4:41 AM, ☽Ţ ҉ĥϋηϠ₡яღ☯ॐ€ðяئॐ♡Pϟiℓℴϟℴ said:

    Is sulphur very wideley used or maybe considered alternative? if applicable - that one to the list too.



    I used sulphur burners on a timer with good efficacy in commercial planthouses. It's not 100% but it did reduce the amount of spraying I needed to do by 80%. Sulphur gas is toxic tho, you need to be sure once your closed room is treated the gas vents safely somewhere no-one is going to breathe it ( ie- innocent bystanders ) , and vented entirely before you enter the room.


    There may be species sensitive to the topical application of sulphur- I never worked with any. Over time ( a decade ) it may degrade some metals, like plant stands, slightly


    The presence of residual sulphur on plants may negatively impact some experiments, but so will a lot of other sprays ( residual fungicides and embryo work in some species is anecdotally one of these ). For TC parent explants everything is washed off prior to culture anyhow so that's not an issue here


    Look, nothing beats good planning, cleanliness and getting onto things quickly before pathogens go exponential ( which happens over about 36 hrs under optimal conditions for the pathogen )


    It sounds too simple to be true and is often ignored. I did TAFE chemcert recertifications a number of times and they did emphasise this heavily as a preferred solution to pest management and they were spot on. Once I started Integrated Pest Management strategies my spraying times went to about 20% of previous


    A crowded plant space with a dirty floor, a clogged drain, poor airflow and overly nutritious propagation mix for your species is a bug buffet, and much of your effort will need to be redirected into sorting out pest problems.


    Also: Trichoderma. I know I rave about that shit, but the plant immune response is ( empirically ) worth it for me. At least I know which pathogen I'm fighting off at sterilisation time too.


    Double sided yellow sticky tape over anything which touches the floors or leads onto the benches. Like this:




    The yellow attracts some bugs, and the sticky prevents them climbing. Easy to manually inspect, if you see a bunch of bugs adhering to the trap, you have an entry point for them. Work out where it is and close the loophole. Inspect the plants for signs of infestation ( if the tape gets overloaded it will lose it's stickiness and become a ladder for more bugs )


    Keep your plants from touching the walls or floors when using these, and wrap tape around any power leads which could let bugs walk onto your babies ( easy to miss if you are running heat mats )

    • Like 1

  14. On 11/24/2017 at 9:17 PM, shonman said:

    i would really like to hurry up and get a disinfected culture into micropropagation tech ASAP.

    but, of course, it must be free of pests, and vigorous...

    also, I dislike cutting the new growth, when the plant needs more leaves!


    That's the spirit!


    And yeah, that last bit sucks. It's hard to resist that temptation I know, but in the long run it's sooooo much better for both your work and the plant.


    Taking parent material from weak plants rarely results in a win for either of you


    Hey I can't remember, but were you initiating new cultures into 1/2 strength media with low sugar? Always a good start, low ( 50% or less ) sugar doesn't feed contaminants anywhere near as much as full strength. Gives your species time to recover.


    I've always believed explants retain some residual immune system after excision ( this is just a belief- I've never looked deeply into whole-plant immune response ). I've seen a few species carry residual contamination for yeeeeeeears without effect as long as there is PPM in the media- and have that contam re-emerge a decade later in the absence of PPM ( positive controls were in place, so yes, it's a thing ). PPM is a biotstat- not a biocide.


    I no longer always run batches of media with PPM, it's exxy. And while I was working this out I did lose batches of plants, so I got cautious and decided PPM needed to go into everything. Then I isolated the compulsory PPM requirements to a few species only, and also always keep a few of every species in PPM in case of equipment or operator error.

    • Like 2

  15. Experiment abandoned.


    I can get a kilo of Trich ferts for about $60, and if I aliquot it under sterile conditions that'll last for 2-3 years


    I don't need the bacto elements in the commercial blends, previous experiences with different commercial fungal/bacto blends have shown some deleterious consequences under some weather conditions with my target species ( long story snipped ).


    Have worked successfully with Trich nutes in these species previously- a fungal blend is overall better but I never did comparisons at a level to determine which one- I just know that without the Trich added to ferts the results were much poorer and I have the dead plants to prove it. The Trich is the constant


    Is gunna be quicker to get $60 together than it is to work out whether the home Trich LC mix is set back/ viable due to shock when diltued into non-sterile working strength solution, and whether that solution is producing comparable outcomes wrt plant growth


    Plus I'd need to buy the Trich nutes anyhow to run as a positive control

    • Like 1

  16. So glad to hear tickets news, this is gunna be a corker. So excited.


    Thank you to the whole EGA team for your level of professionalism and commitment to the event. I'm seriously impressed by the amount of work, passion and consideration going into this.


    If you're sitting on the fence about a ticket like Ronny said I reckon you'd be insane to miss this. Something for everyone, every level of knowledge and interest in the fields. Everyone.

    • Like 4

  17. 1 hour ago, trucha said:

    There is no peyote farming involved in sacrament production for legal users of peyote in the USA or Mexico.

    All peyote is presently wild harvested.


    Thanks heaps for the clarification, it def adds weight and meaning to the numbers


    And thank you again for all your excellent, quality, long-term dedication to the species and issues around them. Brilliant.

    • Like 1

  18. 5 hours ago, Crop said:

     I use Psudomonas for similar results.


    Nice- must try that sometime


    5 hours ago, Crop said:

    I haven't used Trichoderma before but, being a fungus, your plan on upscaling in a sterile liquid sounds like the right way to go. I assume the malt is because you plan on giving the trich a chance to colonise before spraying?


    I'm prolly not thinking this through properly, but I figure the commercial products w encapsulation directly penetrate leaf surface- once sprayed on they germinate. I have no idea if this is accurate.


    Spraying live unprotected log phase colonies onto plants even in low light might not be as effective as the light shock, sudden exposure to air etc. could kill them


    Might work straight into soil tho. Hmmmm...



    4 hours ago, Crop said:

    One possable, simple experiment could be cups of nutrient solution in a controled enviroment, with the only variable between cups being the concentration and preperation of the trichoderma. Float a single Lemna minor in each cup and count how much it multaplies over a week or 2. This will give you a baseline on if Trichs effect plant growth in isolation. More important will be how it reacts with other organisms in your garden. for this you could culture several dishes of your local slime mold and see what it takes to kill it. Maybe you need to add another strain that can handle the cold so you don't need to keep reaplying?


    Duckweed... nice idea, thank you! That way I can avoid all those tedious petri dishes and centrifuging and resuspension and optical density measurements and just go straight to the guts of it- ie does it give visible growth results


    I don't mind re-application as that way I can be certain that precious plants will be OK, especially if I can make an ongoing supply for practically nothing. So competitor organisms aren't an issue


    Positive control will be commercial blend treatment. Negative controls will be untreated Duckweed, and the fert blend minus the Trichoderma. Space limitations mean the number of repeats per variable will be limited to 2-3 and I don't want to be running more than 10-15 duckweed containers for more than 2 months.


    Dammit I'll need to randomise their placement to make sure that light differences aren't fudging results :/


    Hmmm, won't have time to do this before EGA.


    Still open to other improvements or teks while we're thinking about it


    Thanks heaps Crop!