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Invisibletahoe
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Acclimating a species
    #3615322 - 01/12/05 12:30 AM (11 years, 10 months ago)

I just recieved an almond portobella kit from fp and they have it growing on a wood/bran mixture. Agaricus usally grow in a dung/straw mixture so i emailed fp and they said that they acclimated this species to grow on wood. Anyone know exactly what they mean my acclimating or how. I was thinking of starting agaricus biporus on a wood/bran mixture and cloning the biggest shrooms. Would this work or should i start with spores and a wood/bran agar and isolate the most vigorous strain


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Offlinedebianlinux
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Re: Acclimating a species [Re: tahoe]
    #3617754 - 01/12/05 02:05 PM (11 years, 10 months ago)

Acclimating generally refers to climatic conditions. IOW, taking a specimen that fruits naturally in Indonesia and "training" it to fruit, naturally, in the US is acclimation.

In your case they are referring to the fact that your particular specimen/strain has been eating wood based substrates for a few generations. While the specimen may very well grow on dung/straw, you would most likely need to grow the strain out on dung/straw for a few generations (either taking clones or multispore) to get yields that compare to or exceed what the original specimen performed on wood. This is because the specimen has been "trained/acclimated/adapted" to wood based substrates. IOW, the mycelium's genetic database is optimised to produce wood-decomposing enzymes as opposed to straw/dung. This is not to say the database (quite literally DNA) doesn't have the information necessary to decompose straw/dung.

Unless you intend to "re-acclimate" this particular "strain" you may do yourself a favor to locate a "strain" that has been growing on dung/straw for awhile.


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InvisibleATWAR
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Re: Acclimating a species [Re: tahoe]
    #3629233 - 01/14/05 06:39 PM (11 years, 10 months ago)

This is interesting.


In reality, I do not believe any "acclimating" has been done in this instance. I think the real work that has been done is that a wood-based substrate (supplemented) has been developed to work with the mushroom strain in question. I highly doubt they have "acclimated" this mushroom to be a wood-decomposer. I think what they have done is more about matching the strain to the substrate and vice versa. Wood based substrates would be much easier to prepare and handle than compost based for such a business, and given the proper supplementation, it seems easily possible. (a friend of mine accidentally grew cubensis out on a wood block made up for Shiitake. This does not mean he acclimated that particular strain to wood, it just happened to fruit off the supplemented block quite well.)


Growing this mushroom out and starting from its spores would be a step in the right direction. There is little doubt you would be able to get an isolate that performs well on compost. Cloning a specimen from this grow will only get you the exact same strain you started with - the one that is supposedly "acclimated" to wood. There is also little doubt in my mind that this same strain will also grow on compost...


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Invisibletahoe
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Re: Acclimating a species [Re: ATWAR]
    #3642175 - 01/18/05 05:07 AM (11 years, 10 months ago)

Quote:

ATWAR said:
This is interesting.


In reality, I do not believe any "acclimating" has been done in this instance. I think the real work that has been done is that a wood-based substrate (supplemented) has been developed to work with the mushroom strain in question. I highly doubt they have "acclimated" this mushroom to be a wood-decomposer. I think what they have done is more about matching the strain to the substrate and vice versa. Wood based substrates would be much easier to prepare and handle than compost based for such a business, and given the proper supplementation, it seems easily possible. (a friend of mine accidentally grew cubensis out on a wood block made up for Shiitake. This does not mean he acclimated that particular strain to wood, it just happened to fruit off the supplemented block quite well.)






Not sure i know what the hell you were talking about. Acclimating was the word fugi perfecti used when i emailed them.


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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OfflineAnnoA
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Re: Acclimating a species [Re: ATWAR]
    #3642548 - 01/18/05 10:30 AM (11 years, 10 months ago)

Bioresource Technology 77 (2001) 65-69

Adapting substrate formulas used for shiitake for production of brown Agaricus bisporus

Jose E. Sanchez a, Daniel J. Royse b

a El Colegio de la Frontera Sur. Apdo. Postal 36. Tapachula, Chiapas 30700, Mexico

b Department of Plant Pathology, Mushroom Research Center, 316 Buckhout Laboratory, The Pennsylvania State University, University Park, PA 16802, USA

Received 31 May 2000; received in revised form 17 August 2000; accepted 26 August 2000

Abstract

A pasteurized, non-composted substrate (basal mixture) consisting of oak sawdust (28%), millet (29%), rye (8%), peat (8%), alfalfa meal (4%), soybean flour (4%), wheat bran (9%), and CaCO3 (10%) was adapted from shiitake culture to produce the common cultivated mushroom (brown; portabello), Agaricus bisporus. Percentage biological efficiency (ratio of fresh mushroom harvested/oven-dry substrate weight, %BE) ranged from a low of 30.1% (when wheat straw was substituted for sawdust) to 77.1% for the basal mixture. Special, high gas-exchange bags were required to optimize mycelial growth during spawn run. Our formula may allow specialty mushroom growers to produce portabello mushrooms on a modified, pasteurized (110°C for 20 min) substrate commonly used for shiitake production without the added expense of compost preparation. © 2001 Elsevier Science Ltd. All rights reserved.

1. Introduction

The first successful demonstration that Agaricus bisporus could be produced on non-composted substrates was reported by Till (1962). He obtained yields that were comparable to conventional compost if a five-step procedure was used that involved the grinding, mixing, sterilizing, spawning and filling of a mixture of 69% straw, 12.3% peat, 9.8% CaCO3, 4.1% soybean flour and 4.1% cotton seed meal. Later, Murphy (1972) reported that it was possible to eliminate phase I composting and obtain respectable yields with only phase II composting if mixtures of sawdust, brewers grain, corn cobs and/or spent compost were used. Finally, Mee (1978) reported that a non-composted mixture of cold manure, peat and lime successfully was used to produce good quality A. bisporus.

In recent years, production of specialty mushrooms on nutrient-supplemented sawdust has increased at a rate of more than 20% annually in the United States (USDA, 1992, 1999). In 1998, for example, shiitake production increased 31% (USDA, 1999). Most production of shiitake is on synthetic media containing sawdust as the basal ingredient (Royse, 1996). Growers typically produce more than one type of specialty mushroom on their farm, but only mushrooms that are adapted to grow on non-composted substrate are available for production. Thus, the increasingly popular brown form ("portabello") of A. bisporus has not been produced using non-composted substrate. Therefore, we wanted to explore the possibility of producing A. bisporus on non-composted substrate using technology developed for shiitake production.

2. Methods

2.1.  Strains

A commercial brown strain (CS800, Sylvan Spawn Laboratories, Kittanning, Pennsylvania) of A. bisporus for the production of "portabello" (Fig. 1) was used.

2.2.  Substrate for mycelial growth

In order to measure the linear extension rate of strain CS800, two experiments were conducted. The first experiment consisted of a mixture of different organic byproducts and supplements as stated in Table 1. A phase II wheat straw-based compost (freshly prepared according to the short method of composting by Sinden and Heuser, 1953) was used as a control. The second experiment consisted of different mixtures of peat/limestone supplemented with various nitrogen-rich compounds as shown in Table 2. Sterilization was completed in an autoclave at 121°C for 1 h. After cooling, six grains of rye spawn were placed in the bottom of a sterile 50-ml centrifuge tube (Beckman) and 25 g of wet sterile substrate was added. The tubes then were incubated at 25°C for 20-25 days. Mycelial growth was observed and measured every two days. At least five tubes per treatment were evaluated.

2.3. Substrate for fruiting

A basic mixture (BM) was defined with ingredients as follows (% based on weight of oven dry ingredients): oak sawdust (28%), millet (29%), rye (8%), peat, alfalfa meal (4%), soybean flour (4%), wheat bran (9%), and CaCO3 (10%). Substituting ingredients such as straw, brewers grain, and corncobs for rye and sawdust modified the BM as shown in Table 3.

2.4. Cultivation methods

Standard cultivation methods for A. bisporus were used (Wuest and Bengston, 1982), except that a non-composted substrate was used. The ingredients were mixed, moistened to 48-55%, thermally treated (110°C, 20 min), cooled and spawned with 0.8-1% rye grain spawn using a paddle mixer previously described (Royse, 1997). After spawning, the substrate was bagged in 2.65 or 6.0 kg sterile bags with a patch filter (Unicorn Import and Manufacturing, Commerce, TX) and then heat-sealed. Temperature during spawn run was maintained at 18-19°C for 2-3 weeks. Bags were opened and 2.5 cm of casing (neutralized peat moss inoculated with mycelium-colonized substrate) was overlaid on the substrate surface (Tschierpe, 1990). Case hold lasted 2-3 weeks at 18-19°C. During case hold, water was applied daily as outlined by Schisler and Wuest (1978).

Table 1

Linear mycelial growth rate of Agaricus bisporus ("portabello", CS800) on five formulas of synthetic substrate and phase II compost (control)

Trt. no.

Ingredients

Mycelial

Wheat straw (%)

Cottonseed hulls (%)

Sawdust (%)

Wheat bran (%)

Peat (%)

Ground soybean (%)

Charcoal (%)

CaCO3

(%)

growth (mm/day)"

1

0

32

32

10

12

4

0

10

2.91b

2

0

37

37

0

12

4

0

10

3.22b

3

37

37

0

0

12

4

0

10

2.22c

4

66

0

0

0

12

8

4

10

1.88c

5

70

0

0

0

12

8

0

10

1.94c

Control

6.18a

a Means followed by the same letter are not significantly different (a = 0.05 after Bonferroni bound). a Standard wheat straw-bedded horse manure phase II compost.

Table 2

Linear mycelial growth rate of Agaricus bisporus ("portabello" CS800) on mixture of peat/lime supplemented with various protein sources

Trt. No.

Ingredients

Mycelial

Yeast culture (%)

Skimmed milk (%)

Rye

(%)

Wheat bran (%)

Peat (%)

Ground soybean (%)

Corn (%)

CaCO3 (%)

growth (mm/day)a

1

0

0

0

0

25

25

0

50

2.76a

2

0

0

0

25

25

0

0

50

2.17b

3

0

0

0

0

25

0

25

50

1.45c

4

0

0

25

0

25

0

0

50

0.38d

5

0

25

0

0

25

0

0

50

0.26d

6

25

0

0

0

25

0

0

50

O.Ole

Control

0

0

0

0

50

0

0

50

1.63bc

'Means followed by the same letter are not significantly different (a = 0.05 after Bonferroni bound).

Table 3

Yield (kg/m ) and percentage biological efficiencies (%BE) of Agaricus bisporus grown (three crops) on basal ingredients and basal ingredients plus or minus various supplements

Crop No.

Treatment description

Yield (kg/m2)

BE (%)

1a

Basal ingredients'5

31.4ac

77.1

Basal ingredients (minus type)

+4% brewer's grain+4% rye

20.9b

51.3

+8% brewer's grain

19.0b

46.5

2a

Basal ingredients

18.7a

47.7

Basal ingredients (minus sawdust)

+ 14% wheat straw+14% sawdust

15.7ab

40.0

+28% wheat straw

11.0b

30.1

Basal ingredients

61.5a

69.2

Basal ingredients (minus rye)

+8% corncobs

60.2a

66.4

aWet (53% moisture) substrate weight = 2.65 kg/bag.

b Basal ingredients were as follows: Oak sawdust (28%), millet (29%), rye (8%), peat (8%), alfalfa meal (4%), ground soybean (4%), wheat bran (9%), and CaCO3 (10%).

c Means followed by the same letter within a crop are not significantly different according to the Waller-Duncan k-ratio (-test (P = 0.05).

d Wet (53% moisture) substrate weight = 6 kg/bag.

2.5. Parameters evaluated

Mycelial growth measurements were plotted against time and the regression line for each treatment was calculated (Statistica, StatsSoft, USA, version 5.5). The slope of each line was denned as the linear extension rate (LER). Total weight of mushrooms per bag (TWM), weight of mushrooms >20 g/mushroom (= total port-abellos or TPM), biological efficiency (BE - ratio of fresh mushroom harvested (kg)/kg dry substrate, expressed as a percentage) and the yield in kg/m2 were determined after the third break.

2.6. Experimental design and statistical analysis

For LER a multivariate comparison of slopes was made (Kleinbaum et al., 1998) applying a correction of the significance level by the Bonferroni procedure (Si-mes, 1986). For fruiting parameters, a completely randomized design with 10 replications was used. The anova and means separation was performed using the GLM procedure (SAS Institute, 1998).

3. Results

3.1. Mycelial growth

The effect of various mixtures on LER of CS800 is shown in Table 1. The LER was 6.18 mm/day on standard phase II compost (control). Treatments one and two, containing a mixture of cotton seed hulls and sawdust had LERs of 2.91 and 3.22 mm/day, respectively. The three treatments containing straw were considered group C containing the lowest LERs (2.22, 1.88 and 1.94 mm/day).

The LERs of different peat/lime mixtures supplemented with various proteinaceous materials are shown in Table 2. The highest value (2.76 mm/day) was obtained when grown soybean was used as a supplement. This treatment formed group A that was significantly different (P = 0.05) from the rest of the treatments. A second group (B) was formed by both the control and the treatment, where wheat bran was used as a supplement (1.63 and 2.17 mm/day, respectively). Although no statistical difference (P = 0.05) was observed in LERs between these two treatments, a difference in mycelial growth pattern was observed. For example, the mycelium in the peat/lime mixture was thin and dispersed while in the wheat bran/peat mixture, and in the soy mixture the mycelium was whiter and denser. The corn-supplemented mixture formed group C with a LER of 1.45 mm/day. The last treatment contained yeast culture, where no growth was observed.

3.2. Biological efficiency

The results obtained when brewer's grain was substituted for rye grain (crop 1) in the BM are shown in Table 3. The highest % BE was obtained with BM (77.1%) and was significantly higher (P = 0.05) than the other two treatments containing brewer's grain (51.3% and 46.5%, respectively).

The effect of increasing levels of wheat straw substituted for sawdust (crop 2) in the BM is shown in Table 3. The highest yield (18.7 kg/m2) was obtained with BM (without wheat straw). There was no significant difference (P = 0.05) between the BM and the treatment where 50% of the sawdust was replaced by wheat straw. Lower mushroom yield was obtained from the treatment where wheat straw completely replaced the sawdust. Mycelial growth was more uniform and more rapid in BM than in the other two treatments.

The effect of substituting corncobs for rye is shown in Table 3 (crop 3). Overall, yields were higher for crop 3 than the other two crops. This was due to the increased amount (more than IX) of substrate (6 kg/bag) compared with crops 1 and 2 (2.65 kg/bag). There was no significant (P = 0.05) difference in yield or BE between treatments when corncobs were substituted for rye in the BM.

4. Discussion

In the United States, production and consumption of brown A. bisporus (portabello) increased 7% from 1998 (21,220 tons) to 1999 (22,771 tons; USDA, 1999). The value of the crop increased 12% (USDA, 1999). The increasing popularity of these mushrooms has paralleled the increasing sales (up 35%o last year) of specialty mushrooms in the United States. Some consumers consider the brown varieties as specialty mushrooms. Thus specialty growers would like to cultivate porta-bellos but may not have compost available. The ability to economically produce portabello mushrooms on non-composted substrate would open the brown market to specialty mushroom growers. It also may benefit certified organic growers since no fungicides or additives are used.

We have demonstrated that it is possible to obtain good yields of the brown A. bisporus on non-composted substrates. According to Schisler (1982) BEs of 50-70%. are considered average for the white form of A. bisporus, while BEs of 70-90%> are considered good. Our yields were as good or better than those obtained from commercial cropping houses in the United States where average yields were approximately 25.7 kg/m (USDA, 1999).

Our observation that yeast culture did not support growth is not well understood since yeast culture contains considerable protein and many essential vitamins and minerals. Also, the reduction in yield when brewer's grain was added to the formula was unexpected since this ingredient is normally used as a nitrogen source in A. bisporus compost (Murphy, 1972). Brewer's grain contains more nitrogen than rye (Stamets, 1993), thus it was expected to produce more mushrooms. Some factor(s) other than nitrogen content may have been responsible for decreased yields since nitrogen contents were 1.45% in BM, and 1.61 and 1.76% in mixtures containing 4% and 8%o brewer's grain, respectively. Brewer's grain may be more important in feeding the microbial population during composting than in supplying nutrients to the developing A. bisporus mycelium.

It is not surprising that A. bisporus grew relatively well on sawdust since Block and Rao (1962) reported that composted sawdust was suitable for producing A. bisporus. Murphy (1972) also used it for producing mushrooms with only phase II composting. The fact that soybean flour supported faster LER in all the peat/ limestone mixtures tested agrees with Sinden and Schisler (1962) who demonstrated that soybean improved mushroom yields. It is interesting to note that although the growth rate observed in SBF/peat/lime-stone mixture was very low (2.76 mm/day) when compared to that obtained with phase II compost (6.18 mm/ day; Table 1), the peat-limestone mixture was selective for A. bisporus. It may be worth exploring the possibility that supplementing peat moss directly with delayed release supplements would be useful for mushroom production.

During the course of our work, we observed that special care must be taken to ensure optimum substrate moisture and to provide optimum aeration inside the bag during spawn run. It was noticed that the greater the amount of substrate contained in the bag, the greater the yield. However, biological efficiency did not increase with increasing substrate quantities. Although we did not conduct specific experiments to determine the optimal size of bags, it was noticed that mycelium did not grow as well in substrate at the bottom of large (6 kg, 25 cm deep) bags (microporus filter patches were present near the top). Thus, it is important that adequate air exchange be provided through appropriate-sized filters. Bags made specifically for this purpose provided sufficient gas exchange and uniform growth from the top to the bottom of the bag.

We also observed that optimum moisture contents in the substrate ranged from 50% to 53%. These levels are considerably below the desired level for phase II compost (around 72%). Thus, additional work regarding substrate moisture contents may improve yields on non-composted substrate.

While our results are promising, substantially more work is needed. Less expensive and more readily available ingredients may reduce the cost of production and make this technology more competitive with compost-produced mushrooms. Such a development would benefit specialty mushroom growers wishing to diversify their product line.

Acknowledgements

The authors would like to thank Tom Rhodes, Doug Keith, and Henry Shawley, from the Mushroom Research Center, PSU and J. Valle from ECOSUR for their kind help in the development of this work. Many thanks are also given to the National Council of Science and Technology (CONACYT, Mexico) for their support during the sabbatical leave of J.E. Sanchez at PSU.

References

Block, S.S., Rao, S.N., 1962. Sawdust compost for mushroom growing. Mush. Sci. 5, 134-141.

Kleinbaum, D.G., Kupper, L.L., Mueller, K.E., 1998. Applied regression analysis and multivariate methods. Ed. PWS-Kent Pub. Co., pp. 276-283.

Mee, H.M., 1978. US Patent 4 127 964.

Murphy, W.S., 1972. Development of a mushroom production medium without phase I composting. Mush. News 20 (12), 4?22.

Royse, D.J., 1996. Yield stimulation of shiitake by millet supplementation of wood chip substrate. In: Royse, D.J. (Ed.), Proceedings of the Second International Conference on Mushroom Biology and Mushroom Products. 9-12 June, University Park, PA, pp. 277-283.

Royse, D.J., 1997. Specialty mushrooms and their cultivation. Hort. Rev. 19, 59-97.

SAS Institute, 1998. SAS User's Guide: Statistics. SAS Institute Statistical Analysis System, Cary, NC.

Schisler, L.C., 1982. New innovations for efficient mushroom growing. In: Wuest, P.J., Bengtson, G.D. (Eds.), Penn State Handbook for Commercial Mushroom Growers. Special Publication, College of Agricultural Sciences, The Pennsylvania State University, 129, pp. 117-118.

Schisler, L.C., Wuest, P.J., 1978. Watering and ventilation from casing through cropping in commercial mushroom production. Special circular 140. The Pennsylvania State University, University Park.

Simes, R.J., 1986. An improved Bonferroni procedure for multiple tests of significance. Biometrika 73, 751-754.

Sinden, J.W., Heuser, E., 1953. The nature of the short composting process and its relation to short composting. Mush. Sci. 2, 123-131.

Sinden, J.W., Schisler, L.C., 1962. Nutrient supplementation of mushroom compost at spawning. Mush. Sci. 5, 267-280.

Stamets, P., 1993. Growing Gourmet and Medicinal Mushrooms. Ten Speed Press, Berkeley, CA.

Tschierpe, H.J., 1990. Cacing, the elegant method to influence crop rhythm. In: Proceedings Australian and New Zealand National Mushroom Industry Conference, Mushroom Growers Association, Australia, pp. 96-104.

Till, O., 1962. Champignonkultur auf sterilisiertem naehrsubstrat und die wiederverwendung von abgetragenem kompost. Mush. Sci. 5, 127-133.

United States Department of Agriculture, 1992. Mushrooms. Agricultural Statistics Board, Washington.

United States Department of Agriculture, 1999. Mushrooms. Agricultural Statistics Board, Washington.

Wuest, P.J., Bengston, G.D. (Eds.), 1982. Penn State Handbook for Commercial Mushroom Growers. Special Publication, College of Agricultural Sciences, Pennsylvania State University, 129p.



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InvisibleATWAR
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Re: Acclimating a species [Re: Anno]
    #3644915 - 01/18/05 08:38 PM (11 years, 10 months ago)

Very nice read anno! Yet another post to add to my personal archives...

Quote:

2.1. Strains

A commercial brown strain (CS800, Sylvan Spawn Laboratories, Kittanning, Pennsylvania) of A. bisporus for the production of "portabello" (Fig. 1) was used






I think it is important to note that a comercially available strain was used for this substrate. There was no mention about adapting, acclimating, or selecting a strain to be productive on the wood based substrate...

The technique is adapted...


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OfflineMycelium5150
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Re: Acclimating a species [Re: ATWAR]
    #3645111 - 01/18/05 09:14 PM (11 years, 10 months ago)

Anno,
Thanks for the info. Tahoe will like that answer


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