Experiments+Strategy

See Experiments Log ** Mushroom Cultivation Strategy ** =**Mission - Tunisia**=

**Operation Explore - 01.2014**

 * **A. Exploring substrates**
 * 1) Sawdust & woodchips : Oak, poplar, acacia, pine & olive
 * 2) Straw standard technique
 * 3) Fructification on grain
 * 4) Coffee ground & misc (olive waste)
 * **B. Mycelium and spore library**
 * PO & PC spore print

**Operation Everywhere - 10.2013**
Expand cultivation and lab to make a larger production. Five new spore syringe (P. pulmonarius, Hypsizygus ulmarius, Stropharia rugoso-annulata, Agrocybe aegerita & Agaricus bisporus). New shiitake commercial dry spawn. The fridge containing spores, especially P. pulmonarius) and cultures has been accidentally frozen the 04.11.13 (button moved inadvertently, now it is taped), thus viability of all cultures is under examination.
 * **A. Fructification maximale**
 * 1) Hypsizygus master grain from previous operation to grain spawn G1 - done 29.10.13
 * 2) Pleurotus pulmonarius spore syringe germination on agar P0 - done 30.10.13 with success for all syringe
 * 3) More Hypsizygus spawn G1 to G2 - done 09.11.13 for 5 jars & 1 bag & 16 - 17.11.13 for 6 jars & 2 bags
 * 4) Hypsizygus test & first batch - done 15.11.13 with 8 kg of straw
 * 5) Pleurotus eryngii grain master - done 17.11.13 for 4 jars
 * **B. Mycelium and spore library**
 * 1) P. ostreatus from bought wild specimen cloned to agar - done 15.12.13
 * 2) P. citrinopileatus, P. eryngii, P.ostreatus & Ganoderma lucidum clones transfer to new agar - done 30.10.13
 * 3) Agaricus blazei-murrill old spore syringe germination - done 30.10.13
 * 4) P. eryngii clone (transferred from donia clone) transfer to new agar - done 30.10.13
 * 5) Test viability of new spore syringes - done 05.11.13
 * 6) Test viability of stored Hypsizygus P0, before transfer to slant - done 05.11.13
 * 7) Saving P.citrinopileatus, operation from all agars to new agar - done 08.11013
 * 8) Test viability of H.U. before slant - done 08.11.13
 * 9) P. eryngii commercial spawn, possibly isolated strain, transfer to agar
 * 10) Lentinula edodes dry commercial spawn transfer to agar
 * 11) Make tubes of current species - done for P.E. 05.11.13, redone for P.E. & G.L. 08.11.13
 * 12) Test old syringes viability
 * **C. Expanding techniques**
 * 1) L. edodes on outdoor apricot log
 * 2) Outdoor bed of P. pulmonarius & Hypsizygus

Operation Restart 05.2013 - 09.2013
Restart cultivation after 1 year of pause, make new fresh material
 * **Test spore syringe viability** : make inoculation test --> 10 agar done, after 5 days, first germination (Hypsizygus)
 * **Unknown spawn : Hypsizygus or Pleurotus ?**
 * 1) identify (fructification on grain), then G2G
 * 2) fructification on a common media : straw --> 2, 10 & 4 bottles with different spawn rate done, waiting result
 * **Pleurotus eryngii spawn**
 * 1) G2G on grain
 * 2) fructification on straw (?)
 * 3) make spore print
 * **Pleurotus djamor dirty prints**
 * 1) germinate on agar
 * 2) purify on agar
 * 3) grain spawn
 * **Donia Pleurotus eryngii, citrinopileatus & pulmonarius and Ganoderma lucidum clones**
 * 1) multiply on agar : Made transfers, but huge bacterial contamination. Made new transfers and purification, eryngii survived (07.09.13)

>
 * Starting material **
 * ** Spores **
 * ** Commercial Syringe **from MRCA, from 05.2012 (was kept in a fridge that was mistakenly frozen at 9.2013 & 04.11.13)
 * 2 syringes **Hypsizygus ulmarius** (was found alive in 8.2013)
 * 2 syringes **Pleurotus pulmonarius** (did not germinate in 08.2013)
 * 1 syringe **Stropharia rugoso-annulata**
 * 1 syringe **Agaricus blazei murill**
 * 1 syringe ** Pioppino (Agrocybe aegerita ) **
 * **Home made Spore prints**, from 01.2012
 * dozens prints of **Pleurotus djamor**
 * few prints of **Shiitake**
 * **New spore syringe** from MRCA, from 10.2013, accidentally frozen the 04.11.13
 * 1 syringes ** Hypsizygus ulmarius **
 * 1 syringes ** Pleurotus pulmonarius **
 * 1 syringe ** Stropharia rugoso-annulata **
 * 1 syringe ** Agaricus bisporus **
 * 1 syringe ** Agrocybe aegerita (Pioppino) **
 * ** Home made Agars **, from 06.2012, accidentally frozen the 04.11.13
 * 2 agars petri Hypsizygus ulmarius
 * 3 agars jar unknown (Hypsizygus or Pleurotus)
 * **Donia small agar plates,** from 01.2013, accidentally frozen the 04.11.13
 * P. eryngii, P. citrinopileatus, P. ostreatus & P. pulmonarius
 * Ganoderma lucidum
 * Spawn
 * 1 commercial bag spawn ** Pleurotus ****eryngii** (600gr), 03.2012, accidentally frozen the 04.11.13
 * 1 commercial dry spawn **Lentinula edodes**, 10.2013

Equipment

 * glovebox with positive pressure
 * 2 pressure Cooker 10L & 4.5L
 * mediums
 * Grain : 12 kg of organic whole barley + 3l Vermiculite, Gypsum
 * Straw : 2 wheat straw bale
 * Agar* : 5 packets of 80g + Yeast, Peptone*
 * 1+3/4 tyvek* safety suit
 * 95° alcohol (600ml)
 * danish & tunisian masking tape, tunisian normal & gaffer tape
 * items impossible to buy/find in Tunisia

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