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The Corroboree

Darklight

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Posts posted by Darklight


  1. Right now what I'm doing is drooling. I don't understand 80% of the techspeak, which is entirely normal for a new field for me, so I'm not bothered by that. If I needed something doing in this space right now I'd spend the time because obvs the field is expanding and I need to watch the directions. All I can do at this point is observe, and drool

     

    What excites me is that there are people here with relevant and current skillset who can explain it well and who are sharing information

     

    Would it be possible for 2-3 of you to get together and put something up for the next EGA Garden States? Like a DNA 101 workshop on a single relevant point to us ethno freaks?

     

    A few of us have been working on practical protocols for base-level workshops for the day next May. Light microscopy, a little TLC, and something fungal again. Stuff people want to do at home- and can- but just need a few pointers to get the most out of the tek


  2. On 20/08/2019 at 10:39 AM, Tripyamine said:

    Kinda funny but, there is a legend from ANU that has made a group on a website called protocols.io that has customized extractions people have perfected for heaps of different tissue.

     

     

    If it's the same legend from ANU who was proposing exactly that a few years back, please tell him I said hi? He was a legend even back then, now I reckon he'd be a fucken rockstar

     

    Great site too


  3. I don't understand about 80% of the technical speak of the above, and genetics isn't my field

     

    Am incredibly impressed tho, by how much the skillsets of youse all have progressed :)

     

    It's been nearly 3 years since the OP put up their request. What's changed?

     

    My understanding is that SDN-1 knockouts are now no longer covered by OGTR legislation, but not sure if this covers environmental release as well. Anyone know?

     

    Quite possibly what hasn't changed, and may be going backwards, is public access to a lot of new databases.

     

    And I suspect ( but cannot confirm ) that the human factor is still a big part- quality DNA extraction is hard for some species or plant parts and as Tripyamine says above, a bad result needs to be caught before it proliferates. Add that to the variables of technique and machine and it sounds like it'll be a shitfight for a while yet


  4. On 26/07/2019 at 7:28 PM, Maximt said:

    I think why most mush cultivators use horizontal is because its cost effective. i.e. you can DIY - find a filter, a pump and some ply and away you go, and you can set up a 3-4 meter wall of these for relatively cheap. And if you are doing mass inoculation it makes sense and saves time, you can line up all your bags on the desk, clean them all at once and just go through inoculating.

     

     

    Not IME. Horizontal airflow pushes sterile air towards you so if anything falls off your hands or tools while you're working, it is blown forward, away from any media or tools which might be contaminated by falling spore. Cost effectiveness isn't much to do with your choice, you want horizontal flow so as not to contaminate your work while you're working on it.

     

    Cheap HEPA flow units can be great for most people doing myco work and spending more won't necessarily guarantee success if your technique is poor or your existing unit has issues.

     

     

    On 26/07/2019 at 7:28 PM, Maximt said:

    Vertical flow hoods are generally something that is used in laboratories i medical research facilities. The save space, as the filter and pump don't take up desk space. They are generally enclosed from all sides but 1, and have positive pressure to push air out through the bottom of the table and the 1 opening. But if you need a lot of these, i.e. 3-4 meters, becomes very expensive, especially if they have to be custom built. Added bonuses are they are enclosed from all sides, hence less chance for contams to get in, just 1 opening, and take up less desk space.

     

     

    Nup, not IME. Vertical flows are used where there is a danger to the operator or environment from pathogens, and they're often used- routinely used- in medical/ clinical laboratories where they are testing or researching organisms which can cause disease. Pathology labs, working with blood- or air-borne viruses, Zika, HIV, infected tissue of any kind. Biosecurity labs where they may be testing imported plant tissue for bacteria etc, or extracting DNA/ RNA for further tests.

     

    At least one of the Biosafety cabinet classes does both kinds, for work which is required to be sterile and where the operator and environment both require protection. Air is first pushed through a HEPA filter at the back- then rather than being pushed forwards towards the operator, a second air current near the front of the cabinet works like an air curtain and forces the air downwards before it gets to the operator.

     

    I've worked in one for Biosecurity purposes, it's not my normal cabinet, but they've gotten a lot more specific on classes and subclasses of cabinets these days so the changes over time haven't registered on my radar. I do know I've rarely worked in a Class I cabinet ( fume hood type ) and never in a Class III cabinet ( glove box type )

     

    Happy to be proven wrong, I'm out of touch these days

     

    Read this again: https://www.thermofisher.com/nl/en/home/references/gibco-cell-culture-basics/cell-culture-equipment/laminar-flow-hood.html


  5. That's so good to hear re the costings, about half of everything I've tried ends up costing me money rather than saving it

     

    Coupla decades ago I was raising them for meat from the spares.Like pigeons, you can dry pluck them, so they're pretty much an instant meal

     

    Is not for everyone, I understand, but it's so little extra work and quicker than picking up pizza from town

     

    I definitely now have quail envy

    • Like 1

  6. The eggs are really nice in a salad with avo, maccas, greens and a bit of chilli in the dressing

     

    Yeah was thinking about the bug eating aspect. If I ever get around to building another fort knox for some I'll be putting oyster mushroom buckets on bricks, hopefully the quail will eat any of the tiny flies that the mushroom buckets attract

     

    Are they financially efficient? Reckon you get your moneys worth out in eggs? And are you planning on raising more from the eggs?


  7. I'd go if I was in Melbs for sure :D

     

    Look, if you've ever been to a mushroom tek and feel like you haven't learned enough to be as productive as you like, go to another one!

     

    Mycology's quite the thing to wrap your head around- even with years of plant growing and plant biotech experience it took me a coupla years work to get the gist of the different schedulings and life cycles, spawn implications etc.

     

    Luckily the teks taught these days are way simpler than they were back in my day, and we know which strains are better suited to local conditions and substrates.

     

    But it never hurts to do a basics workshop again if you're struggling ( ID workshops I always always learn new things, I try not to miss those ever )


  8. 15 hours ago, Solipsis said:

    Whatever it is, I would consider it serious.. and in my experience it is not good to just start randomly applying solutions especially ones that are mainly meant to maintain and control problems.

    Focus such a treatment on a decent systemic product and don't start putting peroxide on the roots or combine 5 different things meant for prevention, hoping that it will work because you stacked them..

     

    This. Exactly.

     

    Do not throw more than one treatment on any sick plant at a time unless you have experience with that combination and that species.

     

    Give the treatment time to work ( or fail ) before throwing anything else at it

     

    You might lose your plant anyhow, but you're giving it a better chance in the meantime. And if it fails, you can tell people that specific tek didn't work- for your species and pathogen, in your area. Which will help others

     

    My suggestion of Trichoderma has only worked for me thus far, for pathogens in my area, on cactus I've previously had problems with. I've not yet enquired if the protocol has been tried on cactus in other regions by other people. I'd expect anyone throwing Trichoderma as a new treatment for a sick cactus in another bio-region would regard it as an experimental treatment.

     

    My extensive experience in people telling me what works for them has shown that about 80% of people throw a bunch of random stuff at their sick plants, in an entirely undocumented and cavalier way contrary to any instructions, then pick one thing from the bunch and blame it for the success or failure. It makes for unreliable data.

     

     

     

    15 hours ago, Solipsis said:

     

    Tricky cause while you want to get em out of that soil asap which has *way* too much organic soil in it

     

     

    Yup. Lophs here in NNSW can stand being pulled out of the ground over winter to dry out then repotting, but not sure if I'd recommend that til yours has at least started to heal- or even fully healed

    • Like 1

  9. 18 hours ago, woopwoop said:

    So you apply it to the actual plant?

     

    Yup, as a foliar spray

     

    18 hours ago, woopwoop said:

    Do you use a mostly mineral mix for your lophs (especially if your in a tropical location)?

     

    It's a coarse sharp sand blend, I think it was commercial. Planted it in that pot about 15 years ago

     

    18 hours ago, woopwoop said:

    if so, do you still use Trichoderma in the soil of it as well?

     

    Yup. Mostly via what falls on the soil from the foliar spray. Ideally I apply it once every 1-2 weeks when I foliar spray everything here


  10. Dunno if it'll work in your climate but I throw a commercial Trichoderma solution over anything that looks wounded, mouldy or is prone to it.

     

    Early morning, so it has a chance to dry, absorb, multiply before the cool evening sets in.

     

    I've never been able to grow Lophs outside, nor Mammillaria until I started doing this about 18 months ago. They both always got orange rot 100%.

     

    Last growing season was my first ever successful button cactus year. Even late and cool in the season, when a bird pecked a massive hole in the big Loph it healed fully within 48 hrs, callusing started after 12hr after application. No further damage, no pathogens moved in

     

    IME Trich regular applications also stop larvae if you always respray within the breeding cycle

     

    But that's just me, here. Don't use it as a magic bullet for everything without testing on a few of your babies over a season or two.

     

    Was gunna make up a commercial mix and sell it, but my marketing skills suck. Get yours from a hydro shop. Have also posted on other threads here on Trichoderma


  11. Niggles and I have been looking at microscopy, learning more about it. Um, there's a lot more to it than we thought...

     

    What's become obvious early on in the piece is that my cheapo $50 ebay scope isn't up to the job of looking at stomates and spores. It's not just the magnification, it's the crap optics.

     

    Need a compound scope with illumination, pref LED. Pref with coarse and fine focusing. Magnification to 1000x or more if there's an immersion lens involved ( never used one, no idea how that works ). I wanna see stomates, chromosomes, spores.

     

    Good optics are essential. We'll be attaching a camera to it too. Zeiss. Leica or Olympus lens or similar quality.

     

    If you have a good compound scope at home just sitting round and you'd love to see it get used regularly, gimme a hoi and we'll find a suitable trade for you.


  12. On 23/06/2019 at 12:29 AM, Freakosystem said:

    Oh no! Somebody didn't learn their kingdoms of life...

     

    I'm lichen that one more and more

     

    On 23/06/2019 at 12:29 AM, Freakosystem said:

     

    Has anybody successfully fruited Omohalotus nidiformis? A friend had a good culture going and innoculated many logs as well as filter patch bags as per the morphologically similar Pluerotus species and whilst the mycelium was very aggressive, they never fruited.

     

    Same same.

     

    Though there might be one here trying to fruit on the agar plate ( could also be contam, discolouration from inoculation point tho and never seen this particular effect before )

     

    Will post if it does fruit


  13. How did the microscopy workshop go? Wish I'd been there.

     

    Starting an informal microscopy study run today with another forum member to try and get some basic teks in hand to add to current research. Is way more daunting than I thought. Microscopy, like tissue culture and TLC, is an entire field of knowledge of it's own

     

    First lesson is cleaning our microscopes. It's mouldy in NNSW and even the best stored scopes cop it

     

    We're starting with the cheap-arsed compound scopes we've had for years ( USB camera, ebay ) to see how far we can push their utility before we look at the worth of saving for upgrades.

     

    Tho good optics and lens' are important- I once used the USB camera on a 1920s microscope with immaculate optics and the results were absolutely brilliant- way better than the cheap arsed ebay one I was comparing it to

    • Like 1

  14. Yeah nah, I wouldn't. Says it's a vertical flow cabinet- so clean air is pushed from the top of the cabinet to the bottom

     

    Best flow cabinets for general contamination free cultures IMO are horizontal flow. Clean air is pushed from the back of the cabinet towards you, so if any contaminants are inadvertently transferred during work they will be blown away from the work surface towards the user

     

    Anyone here ever used a vertical flow cabinet for routine sterile culture  transfer work? This link says it's possible, but I've not seen it in practice ever

     

    https://www.thermofisher.com/nl/en/home/references/gibco-cell-culture-basics/cell-culture-equipment/laminar-flow-hood.html

     

     

    • Like 1

  15. 8 hours ago, Glaukus said:

    As long as there's food for the mycelium to consume the patch will continue, it will spread outwards.

     

    And as long as there are no competitor organisms and the humidity and temps remain in the Goldilocks zone for that species etc etc

     

    Actually that's a pretty cool question Zool and I don't think the answer has ever been found for all species. Some form underground storage organs like sclerotia which are basically compressed hyphae and hang there til conditions are right to keep growing, some species are always present as mycelia when they grow in symbiosis with trees, and whatever it is with the pseudosclerotia that some fungus form I do not know at all.

     

     

    • Like 4

  16. On 05/06/2019 at 11:32 PM, Pedro99 said:

    I will leave it be and hope it works itself out.

     

     

    It will. Is it still happening?

     

    On 05/06/2019 at 11:32 PM, Pedro99 said:

    In the meantime I will give the worms a proper burial, I always hated impaling them on hooks when I used to fish. :(

     

     

    Relax and let the fungus do it's work, burying them is overkill and just means you work so different fungus can get them, only you don't have to see it

     

    The fungus will take us all one day, as will the bacteria and all the other little critters. Is how it's sposed to be


  17. 16 hours ago, Humbolt said:

    Recently Ive had discussions with a dishwasher tablet company ( Natures Organics who market themselves as environmentally prudent) and was hoping someone with a grasp on the chemistry could advise me before i reply.

    They individually wrap each tablet with polyvinyl alcohol which they claim is " a type of polymer that is fully water soluble; the component ingredients then being fully biodegradable".  I left a wrapper in a glass for a week and noticed no further degradation beyond a silicone coloured liquid suspended in water.  So i wondered how do naturally occuring enzymes finish the job?  Under what circumstances are they present, ie is the packaging likely to come into contact with these enzymes during its journey.

     

    Came across comments from a polymer chemist who says PVA "is water soluable but not sure of the effects on marine life or water based ecological systems" He goes on to suggest there is an issue with "degradation of the aliphatic polymer backbone.  No naturally occuring enzymes or bacteria are currently known to be able to degrade this structure".

     

    The company claims its 'Earth choice' tabs are individually wrapped because 'they can cause irritation in some people'.

     

     

     

    That's depressing, but an excellent point. I switched to individually wrapped dishwashing tabs in my lab dishwasher cos the unwrapped ones would physically degrade over time, but I'm also aware of any potential impact ( less likely on the actual glassware cos after washing it's rinsed twice before drying ) of both detergent and packaging.

     

    All a manufacturer has to do is show best practice for the recognised and available technologies under legislation. Whether the govt or industry should monitor all degradation products/impact is a non-issue, because they're simply not going to for every product til it crops up in the news. It's finally up to consumers to trust that standards are continuous, transparent and honest

     

    Look, seriously the only way to work this stuff out is to do it ourselves. And even then the best experiment design won't replicate the infinitely complex soil and other environmental co-factors. There's not only bacteria, and enzymes, but also microfauna, fungi, slime moulds and things we probably haven't discovered and can't consider as cofactors yet

     

    If you're passionate about this and related issues it'd be worthwhile to consider degradation products, and set up a working protocol to analyse and share results

     

    A basic ( ie facile, simple, thorough but somewhat incomplete ) experiment design would be trivial, funds for a thorough chemical/ microbial analysis of each sample could be more of an issue, but the biggest issue is the logging over time, the synchronisation of standards so that others could repeat the experiment, data integrity, centralisation and multiply redundant ( ie backed up safely ) storage  and finally publicising results so they can be understood and critiqued by a wider audience

     

    It's also a possible backbone plan for a Roundup ( glyphosate ) experiment

     

    Citizen science. It's something everyone can do. And it means you really give a fuck, not just an FB like

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