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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Losing the battle with bacrteria
#14018052 - 02/24/11 02:32 AM (12 years, 11 months ago) |
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Every time I try making a grain jar I've got bacterial infections within a week. My oyster jar made it 75% before it stalled. I'm washing, soaking, simmering, pc'ing 15-20 psi for 60-90 minutes, but within a week they stink. I've got Tyvek lids on some and regular on others. Any advice would be greatly appreciated. There isn't excessive water most the time, but the top 1" dries out pretty signficantly as well.
Could I be cooking to hot or long? Are the grains not soaked through enough? I don't have many burst grains but they're kinda hard when they go in.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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chains


Registered: 01/31/11
Posts: 323
Loc: Olde blighty
Last seen: 3 years, 7 months
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14018242 - 02/24/11 04:14 AM (12 years, 11 months ago) |
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Ive been having a few probs with my popcorn jars lately as well. Gettin about 30% contam in my jars to Bacilus. I cant figure out why either! I soak and pc just like you...
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ProfessorPinHead
Trapped in the Archives....




Registered: 07/09/10
Posts: 4,568
Loc: F=G*m_1*m_2/r^2
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14018243 - 02/24/11 04:16 AM (12 years, 11 months ago) |
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This is to both you guys.
Are they wide mouth jars?
If so are the rings and lids used?
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chains


Registered: 01/31/11
Posts: 323
Loc: Olde blighty
Last seen: 3 years, 7 months
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Mine arent wide mouth and just have regular lids with a whole and polyfil for a filter.
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ProfessorPinHead
Trapped in the Archives....




Registered: 07/09/10
Posts: 4,568
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Re: Losing the battle with bacrteria [Re: chains]
#14018284 - 02/24/11 04:47 AM (12 years, 11 months ago) |
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Have they been used a couple of times. Sometimes the rings get warped and it can leave small gaps and shit can get into the jars after pc'ing. If you hold the bad jar and run your finger in a circle around the rim of the lid and feel a warble in it then that is likely the issue.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Bad syringe. Agar it.
-------------------- You make it all possible. Doesn't it feel good?
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Sillicybin
Registered: 02/14/05
Posts: 2,134
Loc:
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14018521 - 02/24/11 06:49 AM (12 years, 11 months ago) |
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Quote:
Doc_T said: Bad syringe. Agar it.
This. Also, make sure you're PCing for 90 minutes (you said 60 to 90, but go for 90.) A lot of people get away with 60, but especially if you're not at sea level, the extra time can be necessary.
If you don't have any agar on hand, sterilize a test jar that you do not inoculate and see if it contaminates. That will isolate your sterilization procedure from your inoculum so you know where your point of failure lies.
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The_Outsider
Stranger


Registered: 12/21/10
Posts: 165
Loc: PNW
Last seen: 6 years, 2 months
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Re: Losing the battle with bacrteria [Re: Sillicybin]
#14018613 - 02/24/11 07:35 AM (12 years, 11 months ago) |
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Quote:
Sillicybin said:
Quote:
Doc_T said: Bad syringe. Agar it.
This. Also, make sure you're PCing for 90 minutes (you said 60 to 90, but go for 90.) A lot of people get away with 60, but especially if you're not at sea level, the extra time can be necessary.
If you don't have any agar on hand, sterilize a test jar that you do not inoculate and see if it contaminates. That will isolate your sterilization procedure from your inoculum so you know where your point of failure lies.
yup 90 minutes. Its been said time and time again. 60 is flirting with disaster. Why chump yourself when you have already done all the work?
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tripdawg420
low life with no life



Registered: 02/02/09
Posts: 7,071
Loc: illinois
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Re: Losing the battle with bacrteria [Re: The_Outsider]
#14018754 - 02/24/11 08:36 AM (12 years, 11 months ago) |
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http://www.shroomery.org/forums/showflat.php/Number/11525679 i have used this tek prolly 70 times perfect every time just use hot water thru the whole tek but it could be any thing just use sponsers at first for yur syringes so u can eliminate alot of shit that could go wrong then learn how they do it ya know well good luck
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conformist


Registered: 07/29/10
Posts: 210
Loc: SE
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14018835 - 02/24/11 08:58 AM (12 years, 11 months ago) |
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How are you inoculating you grain? Are you shooting spores through a port in the lid or opening the lid to put them in? Are you putting a piece of colonized agar in the jar?
Are you using a flowhood or Still air Box of some sort while inoculating or are you doing it in open air?
Leave a jar untouched on your next batch and see if it gets contaminated.
I'm sure you'll find the problem.
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chains


Registered: 01/31/11
Posts: 323
Loc: Olde blighty
Last seen: 3 years, 7 months
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Quote:
ProfessorPinHead said: Have they been used a couple of times. Sometimes the rings get warped and it can leave small gaps and shit can get into the jars after pc'ing. If you hold the bad jar and run your finger in a circle around the rim of the lid and feel a warble in it then that is likely the issue.

Yup think its my old jars that are the problem. Ive got contams from agar and from g2g but my aseptic technique's spot on so think its time for me to get new jars.
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RogerRabbit
Bans for Pleasure



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Re: Losing the battle with bacrteria [Re: conformist]
#14018996 - 02/24/11 09:42 AM (12 years, 11 months ago) |
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If the top 1" of grains are drying out, then you don't have a proper lid. A properly made grain jar won't dry out for six to nine months. Tyvek is a piss-poor filter at best for jars, and should never be used as a 'lid'. Post Office tyvek is even worse than coverall tyvek, but neither hold up like synthetic filter disks.
The most common cause of bacteria in grain jars is the breath and/or dirty fingernails of the cultivator. I suspect you're failing to use gloves and a surgical(not dust) mask. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: chains]
#14019014 - 02/24/11 09:45 AM (12 years, 11 months ago) |
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Quote:
chains said: Yup think its my old jars that are the problem. Ive got contams from agar and from g2g but my aseptic technique's spot on so think its time for me to get new jars.
New lids, like RR said. There's nothing wrong with your jars. (I like polyfil better than tyvek. Haven't tried the filter disks yet.)
-------------------- You make it all possible. Doesn't it feel good?
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skwerlee
Stranger

Registered: 02/22/11
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14019033 - 02/24/11 09:50 AM (12 years, 11 months ago) |
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Just a quick thought.. after you put your sub into the jar, before you put your verm barrier on the top. Are you wiping all your edges clean? I have heard of contam happening this way.
better luck in the future mate!
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siko887
Broke


Registered: 01/05/11
Posts: 772
Loc: Ontario, Canada
Last seen: 3 years, 3 months
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Re: Losing the battle with bacrteria [Re: skwerlee]
#14019120 - 02/24/11 10:09 AM (12 years, 11 months ago) |
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Quote:
skwerlee said: Just a quick thought.. after you put your sub into the jar, before you put your verm barrier on the top. Are you wiping all your edges clean? I have heard of contam happening this way.
better luck in the future mate!
These are grain jars. What are you talking about with a dry verm layer? Grain jars have a completely different filter system that normally use polyfill, tyvek or filter discs. There should be no vermiculite introduced at all during this stage.
-------------------- I don't suffer from insanity, I actually rather enjoy it.
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: skwerlee]
#14019161 - 02/24/11 10:21 AM (12 years, 11 months ago) |
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It's been happening on both my widemouth and regular mouth quart jars. No gloves or mask but tons of alcohol and flame sterilizing. My pf style jars colonize better but lately they've been stalling with contams as well. I'm using sponsor cultures. The Tyvek will be destroyed and plastic lids will be purchased. I have 7 filter disks and tons of polyfil as well. How large of a hole would you recommend in a plastic lid and what size and sealant for an injection port? Ally liquid cultures and agar have been getting bacteria as well. The agar is all done in a glovebox that's been sprayed and sealed until use. I was using cattle insemination gloves but have Tyvek sleeves en route. The bacteria colonies look like opaque spots of slime on agar. This has happened on all my grain attempts whether it be WBS or rye or tertricalle (sp?). I do have N95 masks and a box of nitriles I will start wearing when innoculating jars, but most don't even make it that far.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019167 - 02/24/11 10:22 AM (12 years, 11 months ago) |
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I use 1/4" holes in lids, seems to work ok.
-------------------- You make it all possible. Doesn't it feel good?
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RogerRabbit
Bans for Pleasure



Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 4 days
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019244 - 02/24/11 10:40 AM (12 years, 11 months ago) |
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Quote:
fngbronco said: It's been happening on both my widemouth and regular mouth quart jars. No gloves or mask but tons of alcohol and flame sterilizing.
Bingo. Bacteria problem solved. You should be using alcohol and flame sterilization, a flow hood, etc., but gloves and mask are extremely important.
Quote:
fngbronco said:The Tyvek will be destroyed and plastic lids will be purchased. I have 7 filter disks and tons of polyfil as well.
Plastic lids suck. Get metal lids, drill four 1/16" to 1/8" holes in each lid. Place the lid down, then the filter disk, and then screw the ring on top. This keeps your filter disk dry. If you insist on using a syringe, lift the lid and filter up and squirt the spores directly in, obviously in a glovebox or preferably in front of a laminar flow hood.
Quote:
fngbronco said:I do have N95 masks and a box of nitriles.
If they're N95 dust masks, they're useless. They're designed to protect the wearer from contaminants entering his respiratory system. They do nothing to filter the bad breath of the wearer. Surgical masks on the other hand are designed to stop 99% of the exhaled bacteria from the doctor, thus protecting the patient. This is what we should be using. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: RogerRabbit]
#14019272 - 02/24/11 10:47 AM (12 years, 11 months ago) |
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They're the medical N95 masks. I will start to go under the lid rather than through it from here on out. How should I inoculate ones with polyfil (no money for a while) and how tight should it be placed in the holes?
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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heeroyuy00
Sublime


Registered: 09/27/09
Posts: 299
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019429 - 02/24/11 11:24 AM (12 years, 11 months ago) |
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i used that heat gasket injection port tek with tyvek and ever since i switched to wbs jars ive been 100% even when i have extra spawn i close the lid up and it stays clean some how .. i dont know
but rr is probably right get some new jars and a better filter
and sea level is a factor? ... im below sea level whats that mean ? also bad needle
-------------------- "You ever been to a day care center? Its like a fuckin' frat house... everyones sleeping all over the floor and all the girls are running around pulling there shirts up and tryna put everything in there mouth." Good Links
Edited by heeroyuy00 (02/24/11 11:26 AM)
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019445 - 02/24/11 11:28 AM (12 years, 11 months ago) |
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Quote:
fngbronco said: How should I inoculate ones with polyfil (no money for a while) and how tight should it be placed in the holes?
You can shoot past the polyfil puff if you're careful. Or make silicone injection ports. I fold a little puff over, then reach through with piers and pull it. Too tight to stuff by hand.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14019497 - 02/24/11 11:40 AM (12 years, 11 months ago) |
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K sweet! I'll be starting this whole deal over tomorrow! Gonna empty out and wash and pc all my jars so they're all sterile and clean today! Buying new metal lids and rings and drilling those as well. Stupid ass bacteria out smarting me....
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019512 - 02/24/11 11:42 AM (12 years, 11 months ago) |
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Just don't keep shooting with the same dirty syringe. Get some agar while you're out.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14019959 - 02/24/11 01:20 PM (12 years, 11 months ago) |
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I have agar but it grows bacteria as well. The syringes are clean and the myc is strong (took over some trich on one plate). I've had 3 successful agar plates and even the closed sealed ones will contam (possibly from condensation). Once my glove box has the kinks worked out, to where's I can leave an open plate in there for a couple uses and not get contams, then I'll start at it again. I've just washed all my glass in NABC and am pressure sterilizing at 10psi for 15 min. I'm wondering if my bare fingers on the tape I put over the holes may have attributed as well. I've shot up pf jars with the same solutions with no contams.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022874 - 02/24/11 10:21 PM (12 years, 11 months ago) |
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Quote:
fngbronco said: I've had 3 successful agar plates and even the closed sealed ones will contam (possibly from condensation).
No. Contams come from contams. Contams do not come from sterile water. Your pressure cooking procedure is faulty. Review correct technique and practice doing it right.
You should be able to leave an agar plate alone and have it stay sterile. End of story.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14022908 - 02/24/11 10:28 PM (12 years, 11 months ago) |
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I'm sayin the contams come from the glove box. My sterility was lacking there. All the jars are have been cleaned thoroughly and tomorrow will be my glove box nuking day. Gonna make some agar this weekend and pour test plates. What's the best way to clean things before putting them in the glove box, so as to not drag in contams?
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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biologys
Mycologist in Trainning




Registered: 12/21/09
Posts: 4,622
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022915 - 02/24/11 10:29 PM (12 years, 11 months ago) |
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just a simple bleach/water mixture, then soap/water is all I do.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022931 - 02/24/11 10:33 PM (12 years, 11 months ago) |
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You are doing something wrong. Is it a glovebox or still air box? Either way, wash it out with soap and water. Put items in, close it, wait 15 minutes.
Now, wash hands and glove up. Then go to work.
Leave one dish untouched. If it contams, bad PCing. Open one dish in the glovebox, but don't inoculate it. Open one dish, drag you flamed scalpel through it, close it up
It's gotta be you touching something somewhere. Wish you could shoot a movie, we could tell you in a second.
-------------------- You make it all possible. Doesn't it feel good?
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14022941 - 02/24/11 10:34 PM (12 years, 11 months ago) |
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Wait, wait, wait. You said your syringes are clean? You've never grown a clean culture, how do you know?
-------------------- You make it all possible. Doesn't it feel good?
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Mikeallojee
Coolaid smile



Registered: 10/02/09
Posts: 897
Loc: SW WA
Last seen: 10 years, 11 months
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14023313 - 02/24/11 11:49 PM (12 years, 11 months ago) |
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Plastic lids can be difficult but they can also be handy if used correctly. I have begun to flip the lids on the jars. When used upside down you cant beat them. They slide around a bit but once you get used to them they are as easy as any other and cheaper too.
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14023323 - 02/24/11 11:51 PM (12 years, 11 months ago) |
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It's clean in pf teks but the bacteria gets my grain jars before they colonize.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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heeroyuy00
Sublime


Registered: 09/27/09
Posts: 299
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14024104 - 02/25/11 06:13 AM (12 years, 11 months ago) |
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yeah but you said your jar colonized to 75% then stopped? and how do you know they stink? your not opening the jars are you ?
-------------------- "You ever been to a day care center? Its like a fuckin' frat house... everyones sleeping all over the floor and all the girls are running around pulling there shirts up and tryna put everything in there mouth." Good Links
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14024232 - 02/25/11 07:33 AM (12 years, 11 months ago) |
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Quote:
fngbronco said: It's clean in pf teks but the bacteria gets my grain jars before they colonize.
And your agar plates.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14025424 - 02/25/11 01:06 PM (12 years, 11 months ago) |
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I've got Tyvek lids on half and the others you can smell through the nail holes. They also have that slimy shit that sticks to the glass.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14025466 - 02/25/11 01:19 PM (12 years, 11 months ago) |
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Tyvek is not my favorite filter material. As part of changing one thing at a time to find the problem, grab a bag of polyfil and try it, see what you think. But I don't think that's the problem.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14025691 - 02/25/11 02:12 PM (12 years, 11 months ago) |
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I'll set up the polyfil lids for sure. Heading to the store now!
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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