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Journal of Biology, Agriculture and Healthcare                                               www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012

  Growth Influence of Some Additives on the Mycelial Growth
and Fruit Body Development of Pleurotus Ostreatus (Jacq. Et.
                                         Fr.) Kummer
               Markson, Aniedi-Abasi A.*, Madunagu, B. E., Akpan, U. D., Eshiet, E. I.
              Department of Botany, University of Calabar, P. M. B. 1115. Calabar, Nigeria
                E-mail of corresponding Author: marksoncal@yahoo.com

Abstract
Studies were conducted to establish the response of a wood – rotting fungus, Pleurotus ostreatus to
different growth substrates (sawdust (SD), dry banana leaves (BL) and a combination of both) and
additives [groundnut cake (GC) and palm kernel cake (PKC)]. Spawning (Mycelia running rate) and fruit
body development (area of pileus, length of stipe and fresh weight) were assessed. Initiation of mycelial
growth took place in both treated and untreated substrate samples within 7 days post inoculation (dpi).
Spawn run was delayed in SD substrates treated to PKC beyond 2% and in BL substrates treated with 4%
PKC and above. In BL+SD, spawn run was delayed at all levels of PKC treatments. Spawn run in the BL,
BL+SD and SD controls were completed in 21, 28 and 35 days respectively, following inoculation.
Primordia production occurred in all the samples within 7 days of transfer of bags to the cropping room.
Treatment of substrates with additives (GC and PKC) negatively affected the performance of the substrates.
Addition of 4% of the additives on BL and BL+SD substrates produced mushrooms with smaller pileus
size with the smallest (13.08cm2) observed on 4% PKC-treated BL substrates. Similar trends were observed
in length of stipe, fresh weight and biological efficiency of the mushroom. All substrates evaluated
supported production of fruit bodies for 70 days making a total of 14 flushes from the inception of
cropping.
Key words: Additives, Mycelial growth, fruit body production, Pleurotus ostreatus.
1.        INTRODUCTION
Agro-industry and the wood industry generate enormous amount of wastes yearly which are either burnt or
dumped in places that constitute hazard to humans and the environment. The wise use and recycling of
waste will not only minimize health hazards in the environment but can serve to generate wealth. Chang
and Miles (1989) believe that using only 25% of the yearly volume of burnt cereal straws in the world for
mushroom production could result in a mushroom yield of 317 million metric tons (317 billion kg) of fresh
mushrooms per year. It is also the belief of Poppe (2004) that about 360 billion kg of fresh mushrooms
would have been produced from the estimated 600 billion kg of dry wastes generated from agriculture (500
billion kg) and forestry (100 billion kg) within a year. Such level of mushroom production will directly
translate to 60kg of mushroom per head per year which will automatically provide the 4% protein content
of mushroom to them. Pleurotus ostreatus is a species of the mushrooms belonging to the group commonly
called oyster mushrooms. Like many of the members of the Pleurotaceae, P. ostreatus is a primary
decomposer of hardwood trees and are distributed worldwide (Kong, 2004).
Of the over 70 species of Pleurotus, P. ostreatus is the most important and most commercially viable
because of the compatibility of its growth requirements with the prevailing environmental factors of most
regions of the world. However, different Pleurotus species require varying environmental conditions for
optimal growth. For P. ostreatus, the optimum temperature range for spawn run is put at between 25-350C,
primordia formation occurs at temperature between 10 and 150C while production of fruit bodies is
accomplished                           between                          10                         to170C.
Kong (2004) reported that P. ostreatus requires a sharp drop in temperature usually referred to as ‘cold
shock’ for primordia formation to be initiated. Co2 requirement is put at < 1,000 and the suitable methods
of cultivation ranges from shelf cultivation through cultivation in boxes, in bottles and in bags (Kong,
2004). Substrate is a major component of mushroom cultivation as it forms the nutritional source and the
substratum on which the mushroom is anchored. Among the commercially viable mushrooms known, only


                                                   59
Journal of Biology, Agriculture and Healthcare                                              www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012

Pleurotus spp. have been reported to be capable of utilizing the largest variety of wastes with its fast
mycelial growth and multilateral enzyme system capable of degrading nearly all types of wastes (Poppe,
2004). Worldwide survey on oyster mushroom substrates revealed about 200 different types of substrates
reported to effectively support the growth of Pleurotus spp. Among the listed substrate materials, are dry
banana leaves and sawdust. These materials do not produce optimum mushroom growth in isolation; other
substances are usually added to them to boost their growth-support potentials. Such materials are called
additives. Most additives are organic materials such as flours of maize, wheat, rice, cowpea, horse gram
(Renganathan et al., 2008) and, poultry dung, rice bran, palm kernel cake (PKC), Groundnut cake (GC) and
cow dung (Fasidi, 2006). This study is therefore undertaken to assess two additives for growth performance
of Pleurotus ostreatus on sawdust and banana leaves substrates.

2.       MATERIALS AND METHODS
2.1 Collection of materials

Dry banana leaves were collected from the University of Calabar farms. Sawdust was obtained from
Government-owned timber market at Marian Hill, Calabar. Palm kernel cake was obtained from Federal
Institute of Industrial Research (FIIRO), Oshodi, Lagos State, Nigeria and groundnut cake collected from
hausa traders at Bogobiri, Calabar Municipality, Calabar, Cross River State. The

spawn was donated by Zartech farms, Ibadan, Oyo State, Nigeria. Rice bran was collected from private-
owned rice mill in Itu Mbonuso, Ini L. G. A., Akwa Ibom State, all in Nigeria.

2.2 Substrate composition

Three substrates were composted namely: sawdust, dry banana leaves and sawdust + dry banana leaves.
Sawdust was allowed to ferment for about 30 days. During this period it was turned regularly at 7 days
interval. The fermented sawdust was soaked for 24 hrs. The banana leaves were shredded into bits of about
2cm2 and soaked for 12 hrs. These substrates were transferred to sac bags and drained of water to a
moisture level of 70% using cassava press. Fifty kilograms (50kg) of each of banana and sawdust substrates
were mixed with 0.4kg of CaCO3 and 10kg of rice bran. Pasteurization was achieved in sac bags using
metal drums of about 500litres capacity with a wooden platform of about 35cm high within at 60-800C for
6 hrs (Fasidi, 2006). Initial water level in the drum was 30cm from the bottom of the drum. The pasteurized
substrates were allowed to cool for 12 hrs, and then were divided into 3.5kg portions representing the
content of each mushroom bag. Each of these portions was mixed with 2%, 4% or 6% of either GC or PKC
separately before bagging. Bagging was done in an axenic condition. Plastic bags measuring 30x25cm were
used. Each bag was inoculated with 5g of Pleurotus ostreatus. The open end of each bag was secured with
a PVC pipe 3cm long and 3cm in diameter wrapped with a rubber band and plugged with cotton wool.

2.3 Spawn running

The bags were hanged with ropes serially from the roof down with each line carrying a maximum of five
bags. Spawn running room was allowed limited light. Room temperature was about 300C and relative
humidity between 65 and 75% achieved by spraying the compost bags and walls of the spawn running
room 2 to 3 times daily with clean water (Fasidi, 2006).

2.4 Cropping and harvesting

At the end of spawn run, the bags of mycelial colonized substrates were transferred to the cropping room
stacked on bamboo shelves and opened. This set up was sprayed with clean water after every 6hrs. The
floor of the cropping room was covered with coarse sand from the sea bed and continuously moistened.
The walls of the mushroom house were wetted regularly to humidify the environment to about 70%. On
fruit bodies’ formation and maturity, data were taken on area of pileus, length of stipe using conventional


                                                    60
Journal of Biology, Agriculture and Healthcare                                                  www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012

methods. Fresh weight was taken using Agilent electronic balance. Data were analyzed using SPSS version
14.0.


Biological efficiency (%) = Total weight of the fruit bodies___ X 100
                        Total weight of substrate (compost)     1

3.       RESULTS
3.1 Spawn run
Result from the study revealed that initiation of mycelial growth took place in both treated and untreated
substrate samples within 7 days post inoculation (dpi). However, completion of spawn run was delayed in
SD substrates treated to PKC beyond 2% and in BL substrates treated with 4% PKC and above and in
BL+SD at all levels of PKC treatments. Spawn run in the BL, BL+SD and SD controls were completed in
21, 28 and 35 days respectively, following inoculation (data not shown). The uniformity in the initiation of
mycelial growth in all the substrates may stem from the low hydrolase activity reported of P. ostreatus
during substrate colonization (Elisashvili et al., 2008). The implication is that the level of breakdown of
substrate materials for the release of nutrients is low or non-existent within this short period of 7 days,
hence nutrients in the additives may have not been released and consequently not
available           to          the            mycelia          and           hence           had           no
influence on the initial growth of the mycelia. The variation in the spawn run rate between the treated
substrate samples may have resulted from the differences in the chemical composition of both the
substrates and additives and the rate at which these materials were hydrolyzed. Guillen-Navarro et al.
(1998) linked the production of mycelial biomass to the hydrolytic enzymes and availability of glucose in
the substrate of cultivation. Variations in the speed of colonization (spawn run) by mycelia of different
species of Pleurotus cultivated on different substrates supplemented with various additives have been
reported (Stanley and Awi-Waadu, (2010), Renganathan et al., 2008, Pathmashini et al., 2008). However,
the negative correlation observed between additive supplementation of substrates and the mycelial growth
of P. ostreatus is in contrast with an earlier report (Renganathan et al., 2008) on impact of additives on the
growth of mushrooms. The contrast noted may be a function of the type and availability as well as quantity-
dependent interactions between the nutrient content of the additives and that of the substrates. Chemical
content of palm kernel cake (PKC) revealed the presence low amounts of nutrients and particularly,
proteins reported to be lowest in PKC among the seed cakes in Sub Saharan Africa (FAO, 1984). This may
have contributed to the reduced growth performance on PKC supplemented substrates.

3.2 Cropping
Primordia production in all the substrates took place within the first seven days of transfer of the bags to the
cropping room, however, the earliest (4 days) was in the controls of the three substrates and the last (7
days) to produce primordial was in PKC- treated SD substrates. There were a total of 14 flushes within 10
weeks          with         a        significant        decline           in        growth           parameters
from the 8th week of harvest, especially in mushrooms cultivated on PKC-treated SD substrates.
Assessment of the influence of substrates and additives on the pileus (cap) area of the test mushroom
revealed that areas of mushroom cap were larger in untreated substrate samples compared with those
produced on treated units (Table 1). The largest pileus area (40.64cm2) was recorded in mushrooms
produced on BL substrates (control), while SD substrates (control) supported mushrooms with the least
pileus area (24.82cm2). The disparity observed in the size of pileus produced on BL and SD substrates is
likely a function of nutrient availability from the materials of the two substrates. Sawdust contains high
amounts of lignin (a complex carbon source highly resistant to enzymatic degradation) than cellulose (a
compound easily broken down to glucose which is the form taken in by the fungus as nutrient) and vice
versa (Choi, 2004), hence glucose may have been readily available to the fungus grown on banana leaves
substrates than those on sawdust. However, there were no significant difference (P<0.05) in pileus areas on
SD substrates treated to either additives at all levels of concentration. Generally, addition of 4% of the
additives on BL and BL+SD substrates produced mushrooms with smaller pileus size with the smallest


                                                      61
Journal of Biology, Agriculture and Healthcare                                                www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012

(13.08cm2) observed on 4% PKC-treated BL substrates. This negative impact of additive on pileus size is
probably consequent upon its influence on the mycelial growth (Baysal et al., 2006).
When the performance of the additives were considered irrespective of the level of concentration, it was
discovered that PKC supplemented sawdust substrates recorded a slightly significantly (P<0.05) larger
pileus area than those grown on other substrates. This suggests that, concentration of PKC was the major
determinant of the vigor of growth when added to sawdust substrates. BL and BL+SD substrates performed
better when they were supplemented with GC+PKC. This result points to the fact that the presence of GC
may                                                                                                       have
improved on the nutrient status of the substrate. Groundnut has been reported to be high in nutrients such as
protein (25.2%), oil (4.8%) and 4.5% soluble sugars (Jambunathan, 1991). The additives did not have any
significant effect on the length of stipe, irrespective of the substrate used except marginal differences
observed in mushrooms produced on PKC supplemented SD substrates and in BL substrates supplemented
with zero and 2% GC+PKC (Table 2 ).
There were no significant differences observed in the length of stipe produced on the test substrates
supplemented with different levels of concentrations of the two additives. However, the length of stipe
produced on all the treated substrates were significantly (P<0.05) lower than those produced on untreated
substrate samples (Table 4). This suggests that the growth and development of stipe was not influenced by
the concentration levels of the additives, but by the very presence of the additives in the substrates.
The influence of additives and substrates on the fresh weight of the test mushroom was assessed. A
significant (P<0.05) decline in the fresh weight of P. ostreatus was observed when PKC was added to
sawdust and when the same substrate was supplemented with GC+PKC beyond 2%. The decline in growth
observed seems to be influenced by the introduction of PKC. The low protein status of this supplement may
be the determining factor. FAO (1984) reported PKC to contain the lowest protein level among the oil seed
cakes in Sub-Saharan Africa. On banana leaves substrates, the highest fresh weight of the test mushroom
was recorded on the untreated samples. Treatment with additives did not encourage fresh weight
accumulation in the mushroom cultivated on BL alone. However, when this substrate (BL) was
supplemented with sawdust (BL+SD) and treated with PKC or GC+PKC, the mushroom recorded weight
increases at all levels of the additives concentration (Table 5). The banana
leaves sawdust composite (BL+SD) seems to be compatible with the additives when compared with
sawdust or banana leaves alone. The reason for positive growth support compatibility of the additives with
BL+SD substrates is likely resultant from the high cellulose content of banana leaves (which makes up for
the highly lignolytic nature of sawdust) (Choi, 2004), and the high nutritional content of groundnut cake
(FAO, 1984) which makes up for the low nutrient status of PKC. The difference between the mean fresh
weight of mushroom produced on untreated (no additives) samples and those obtained from additive-
supplemented substrates were in the range of 1.12 to 14.35g (Table 6). The weight differences were smaller
in samples containing BL and GC but higher in sawdust-containing samples treated to PKC alone as the
additives. The result obtained for fresh weight is as expected since the fruit body yield is largely dependent
on the performance of the fungus at mycelial stage and primordia formation (Baysal et al., 2006).
Irrespective of the concentration of the additives, the mean fresh weight of mushroom produced on sawdust
treated to PKC (21.74g) and GC+PKC (22.21g) at the end of 10 weeks of harvest were not significantly
different (P<0.05). However, the mean fresh weight produced on banana leaves substrate treated to
GC+PKC was significantly (P<0.05) higher than the value obtained for samples treated with only PKC.
Similar trend was observed in banana leaves + sawdust substrates (Table 7). The highest mean fresh weight
(29.87g) of mushroom was recorded on banana leaves + sawdust substrates treated with GC+PKC while
the lowest mushroom weight was seen on the sawdust substrate treated to GC+PKC.
Biological efficiency (BE) of the three substrates treated with PKC and GC+PKC were assessed. Results
showed that mushrooms grown on substrate samples without additives treatment recorded higher mean
values of biological efficiency compared with values obtained on treated samples except for BL+SD
substrates which recorded comparable BE values at all levels of additives treatment . Sawdust substrates
treated with 2% GC+PKC and 6% PKC still recorded higher BE values. Generally, sawdust or banana
leaves substrates supplemented with 4% and above of PKC were not efficient in the cultivation of P.
ostreatus (Table 8).


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Journal of Biology, Agriculture and Healthcare                                                 www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012

All growth parameters of Pleurotus ostreatus negatively correlated with PKC and GC + PKC supplemented
sawdust substrates. Sawdust substrates were more biologically efficient when they were supplemented with
2% GC+PKC. Sawdust or banana leaves substrates supplemented with 4% PKC and above were not
efficient in supporting the growth of P. ostreatus. Though, substrates were not supplemented with GC only,
it is likely that supplementation of sawdust and banana leaves substrates alone or BL+ SD composite with
GC alone may produce better results. Substrate supplementation with organic and/or inorganic substances
often boosts mushroom production (Quimio et al., 1990, Renganathan et al., 2008, Adejoye and
Mesewonrun, 2009). This is, however, true but with some reservations. The findings of this study revealed
that substrate supplementation can only be result-oriented when the nutrient status of both the substrates
and the supplement (additive) is considered complementary, and the additives’ concentrations adequate.
This fact should be borne in mind by mushroom farmers, especially when PKC is the additive and sawdust
or banana leaves the growth substrates.

References
Adejoye, O. D., Mesewonrun, O. (2009). The growth and yield of Lentinus squarrosulus (M.) Singer a
          Nigerian edible mushroom as affected by supplements. The Internet Journal of Nutrition and
          Wellness. Volume 8, Number 2.
Baysal, E., Peker, H., Yalinkilic, M. K., Temiz, A. (2003). Cultivation of oyster mushroom on waste paper
          with some added supplementary materials. Bioresource Technology. 89(1):95-97.
Chang, S. T., Miles, P. G. (1989). Edible mushroom and their cultivation. Florida.CRC Press.
Choi, K. W. (2004). Shelf cultivation of oyster mushroom-with emphasis on fermentation. In Mushroom
          Growers’ Handbook. Mushworld. Pp. 153-165.
Elisashvili, V., Kachlishvili, E., Penninckx, M. J. (2008). Lignolytic enzymes profile during growth and
          fruiting     of    Pleurotus      ostreatus     on    wheat       straw     and     tree     leaves.
          Acta Microbiol. Immunol. Hung. 55(2): 157-168.
Fasidi, I. O. (2006). Substrate sourcing and preparation for mushroom cultivation. Training workshop on
          cultivation                                        of                                      tropical
          mushrooms 20th -22nd November, 2006 at University of Ibadan, Oyo State.
Guillen-Navarro, G. K., Marquez-Rocha, F. J., Sanchez-Vazquez, J. E. (1998). Production of biomass and
          lignlytic enzymes py Pleurotus ostreatus in submerged culture. Rev. Iberoam Micol. 15(4): 302-
          306.
Jambunathan, R. (1991). Groundnut quality characteristics. In Uses of tropical grain legumes: Proceedings
          of a Consultants Meet ing, 2 7 - 3 0 Mar 1989, ICRISAT Center, India. Patancheru. Pages 267-
          275. A.P. 502 324, India: International Crops Research Institute for the Semi -Arid Tropics.
Kong, W. (2004). Description of commercially important Pleurotus species. In Mushroom Growers’
          Handbook. Mushworld. Pp. 55-61.
Pathmashini, L., Arulnandhy, A., Wijeratnam ,R. S. W. (2008). Cultivation of oyster mushroom (Pleurotus
          ostreatus) on sawdust. Cey. J. Sci. 37(2): 177-182.
Poppe, J. (2004). Agricultural wastes as substrates for oyster mushroom. In Mushroom Growers’
          Handbook. Mushworld. Pp. 76-85.
Quimio, T. H., Chang, S.T., Royse, D. J. (1990). Technical guidance for mushroom growing in the tropics.
          FAO publication. PP. 1-31.
Renganathan, P., Eswaran, A., Balabaskar, P. (2008). Effect of various additives to the bed substrate on
          the sporophore production by Pleurotus flabellatus (Berk. And Br.) Sacc. Mysore J. Agric. Sci.
          42(1): 132-134.
Stanley, O. H., Awi-Waadu, G. D. (2010). Effects of substrates of spawn production on mycelial growth of
          oyster mushroom species. Res. J. Appl. Sci. 5(3)161-164.




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Journal of Biology, Agriculture and Healthcare                                                     www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012


Table 1: Influence of substrates and additives on area of pileus of Pleurotus ostreatus.



                              Substrate (kg) /Additives (g) and Area of Pileus (cm2)
                                  SD                    BL                    BL+SD
Concentration (g/kg)        PKC     GC+PKC PKC             GC+PKC        PKC        GC+PKC

0                           24.82      24.82       40.64     40.64         35.58      35.58


2                            17.48       24.49      28.36      27.12         16.61         23.73

4                            27.27       27.86      13.08      23.99         19.72         19.70

6                            28.32       19.48      14.32      23.67         18.77         23.44

LSD                            1.24                  1.17                     1.34


SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.




Table 2: Comparative influence of PKC and GC+PKC supplemented Sawdust, Banana leaves and
        Banana leaves + Sawdust substrates on the area of pileus of Pleurotus ostreatus.

                                      Substrate (kg) and Area of Pileus (cm2)

Substrates                     SD                            BL                      BL+SD
PKC                           24.38*                        22.78                     22.13

GC + PKC                      24.16                         28.86*                    25.61*

t- Value                       0.33                          0.41                          0.36

*Significantly different at P<0.05.
SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.




                                                     64
Journal of Biology, Agriculture and Healthcare                                                 www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012


Table 3: Influence of substrates and additives on length of stipe of Pleurotus ostreatus.


                        Substrate (kg) /Additives (g) and Length of Stipe (cm)

                              SD                     BL                 BL+SD
                        PKC        GC+PKC      PKC        GC+PKC      PKC   GC+PKC
Concentration (g/kg)
0 (untreated)             2.66          2.66      3.33       3.33        3.27       3.27

2                          2.10         2.74      1.84       3.30         2.45      2.66

4                          2.00         2.01      1.73       2.38         2.53      2.72

6                          2.00         1.89      2.41       1.97         2.44      2.29

LSD                        1.21                   1.03                    1.15



SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.



Table 4: Comparative influence of PKC and GC+PKC supplemented Sawdust (SD), Banana leaves (BL)
and Banana leaves + Sawdust (BL+SD) substrates on the length stipe of Pleurotus ostreatus.
                          Substrate (kg) and Length of Stipe (cm)

Substrate                     SD                           BL                       BL+SD
PKC                           2.19Ns                      2.33                       2.67Ns

GC + PKC                         2.33                      2.75Ns                       2.74

t- Value                         0.21                      0.43                         0.11


SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Ns= no significant
difference between treatments.




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Journal of Biology, Agriculture and Healthcare                                                            www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012




Table 5: Influence of substrates and additives on fresh weight of Pleurotus ostreatus.

                                   *Substrate (kg) /Additives (g) and Fresh weight (g)

                                            SD                             BL              BL+SD
                                   PKC           GC+PKC        PKC              GC+PKC   PKC    GC+PKC
 Concentration (g/kg)
 0                                  27.87         27.87            33.75         33.75   31.17       31.17

 2                                  13.91         30.69            15.80         24.12    30.34          35.12

 4                                  21.08         16.67            19.92         15.86    37.38          21.30

 6                                  15.74         13.59            12.46         26.69    25.30          31.89

 LSD                                  1.36

*values are means of three determinations
SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.




Table 6: difference in mean fresh weight of P. ostreatus produced on additive-treated and untreated
mushroom substrates.
                                                Fresh weight (g)
Substrates               Untreated                Treated                  Difference
SD alone                 27.87                    -
SD+PKC                   -                        22.07                      5.8
SD+GC+PKC                -                        22.25                      5.62
BL alone                 33.75                    -
BL+PKC                   -                        19.40                      14.35
BL+GC+PKC                   -                     25.11                        8.64

BL+SD alone                31.17                               -
BL+SD+PKC                     -                                    25.72                          5.45

BL+SD+GC+PKC                   -                                   30.05                          1.12

SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.




                                                          66
Journal of Biology, Agriculture and Healthcare                                                  www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol 2, No.3, 2012


Table 7: Comparative influence of PKC and GC+PKC supplemented Sawdust (SD), Banana leaves (SD)
and Banana leaves + Sawdust (BL+SD) substrates on the fresh weight of Pleurotus ostreatus.
                                                Substrate (kg) and Fresh weight (g)

  Substrate                              SD                        BL                 BL+SD
  PKC                                  19.65ns                   20.48                 31.03

  GC + PKC                              18.461                    25.02*                29.87*

  t- Value                               1.22                      1.31                  1.14

*significantly different at P<0.05; ns – not significantly different at P<0.05.
SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.


Table 8: Biological efficiency of Pleurotus ostreatus as influenced by different substrates and additives
                                              *Biological efficiency (%)
                                        SD                       BL                     BL+SD
                                 PKC      GC+PKC          PKC       GC+PKC         PKC        GC+PKC
 Concentration (g/kg)
 0                               7.47     7.96            9.12      9.64           7.71       8.91
 2                               3.97     8.77            4.51      6.89           8.67       10.03
 4                               6.02     4.76            5.69      4.53           10.68      6.09
 6                               3.88     7.35            3.56      7.63           7.23       9.11
*Values are computed from means of 14 flushes.
SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake.




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11.growth influence of some additives on the mycelial growth

  • 1. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Growth Influence of Some Additives on the Mycelial Growth and Fruit Body Development of Pleurotus Ostreatus (Jacq. Et. Fr.) Kummer Markson, Aniedi-Abasi A.*, Madunagu, B. E., Akpan, U. D., Eshiet, E. I. Department of Botany, University of Calabar, P. M. B. 1115. Calabar, Nigeria E-mail of corresponding Author: marksoncal@yahoo.com Abstract Studies were conducted to establish the response of a wood – rotting fungus, Pleurotus ostreatus to different growth substrates (sawdust (SD), dry banana leaves (BL) and a combination of both) and additives [groundnut cake (GC) and palm kernel cake (PKC)]. Spawning (Mycelia running rate) and fruit body development (area of pileus, length of stipe and fresh weight) were assessed. Initiation of mycelial growth took place in both treated and untreated substrate samples within 7 days post inoculation (dpi). Spawn run was delayed in SD substrates treated to PKC beyond 2% and in BL substrates treated with 4% PKC and above. In BL+SD, spawn run was delayed at all levels of PKC treatments. Spawn run in the BL, BL+SD and SD controls were completed in 21, 28 and 35 days respectively, following inoculation. Primordia production occurred in all the samples within 7 days of transfer of bags to the cropping room. Treatment of substrates with additives (GC and PKC) negatively affected the performance of the substrates. Addition of 4% of the additives on BL and BL+SD substrates produced mushrooms with smaller pileus size with the smallest (13.08cm2) observed on 4% PKC-treated BL substrates. Similar trends were observed in length of stipe, fresh weight and biological efficiency of the mushroom. All substrates evaluated supported production of fruit bodies for 70 days making a total of 14 flushes from the inception of cropping. Key words: Additives, Mycelial growth, fruit body production, Pleurotus ostreatus. 1. INTRODUCTION Agro-industry and the wood industry generate enormous amount of wastes yearly which are either burnt or dumped in places that constitute hazard to humans and the environment. The wise use and recycling of waste will not only minimize health hazards in the environment but can serve to generate wealth. Chang and Miles (1989) believe that using only 25% of the yearly volume of burnt cereal straws in the world for mushroom production could result in a mushroom yield of 317 million metric tons (317 billion kg) of fresh mushrooms per year. It is also the belief of Poppe (2004) that about 360 billion kg of fresh mushrooms would have been produced from the estimated 600 billion kg of dry wastes generated from agriculture (500 billion kg) and forestry (100 billion kg) within a year. Such level of mushroom production will directly translate to 60kg of mushroom per head per year which will automatically provide the 4% protein content of mushroom to them. Pleurotus ostreatus is a species of the mushrooms belonging to the group commonly called oyster mushrooms. Like many of the members of the Pleurotaceae, P. ostreatus is a primary decomposer of hardwood trees and are distributed worldwide (Kong, 2004). Of the over 70 species of Pleurotus, P. ostreatus is the most important and most commercially viable because of the compatibility of its growth requirements with the prevailing environmental factors of most regions of the world. However, different Pleurotus species require varying environmental conditions for optimal growth. For P. ostreatus, the optimum temperature range for spawn run is put at between 25-350C, primordia formation occurs at temperature between 10 and 150C while production of fruit bodies is accomplished between 10 to170C. Kong (2004) reported that P. ostreatus requires a sharp drop in temperature usually referred to as ‘cold shock’ for primordia formation to be initiated. Co2 requirement is put at < 1,000 and the suitable methods of cultivation ranges from shelf cultivation through cultivation in boxes, in bottles and in bags (Kong, 2004). Substrate is a major component of mushroom cultivation as it forms the nutritional source and the substratum on which the mushroom is anchored. Among the commercially viable mushrooms known, only 59
  • 2. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Pleurotus spp. have been reported to be capable of utilizing the largest variety of wastes with its fast mycelial growth and multilateral enzyme system capable of degrading nearly all types of wastes (Poppe, 2004). Worldwide survey on oyster mushroom substrates revealed about 200 different types of substrates reported to effectively support the growth of Pleurotus spp. Among the listed substrate materials, are dry banana leaves and sawdust. These materials do not produce optimum mushroom growth in isolation; other substances are usually added to them to boost their growth-support potentials. Such materials are called additives. Most additives are organic materials such as flours of maize, wheat, rice, cowpea, horse gram (Renganathan et al., 2008) and, poultry dung, rice bran, palm kernel cake (PKC), Groundnut cake (GC) and cow dung (Fasidi, 2006). This study is therefore undertaken to assess two additives for growth performance of Pleurotus ostreatus on sawdust and banana leaves substrates. 2. MATERIALS AND METHODS 2.1 Collection of materials Dry banana leaves were collected from the University of Calabar farms. Sawdust was obtained from Government-owned timber market at Marian Hill, Calabar. Palm kernel cake was obtained from Federal Institute of Industrial Research (FIIRO), Oshodi, Lagos State, Nigeria and groundnut cake collected from hausa traders at Bogobiri, Calabar Municipality, Calabar, Cross River State. The spawn was donated by Zartech farms, Ibadan, Oyo State, Nigeria. Rice bran was collected from private- owned rice mill in Itu Mbonuso, Ini L. G. A., Akwa Ibom State, all in Nigeria. 2.2 Substrate composition Three substrates were composted namely: sawdust, dry banana leaves and sawdust + dry banana leaves. Sawdust was allowed to ferment for about 30 days. During this period it was turned regularly at 7 days interval. The fermented sawdust was soaked for 24 hrs. The banana leaves were shredded into bits of about 2cm2 and soaked for 12 hrs. These substrates were transferred to sac bags and drained of water to a moisture level of 70% using cassava press. Fifty kilograms (50kg) of each of banana and sawdust substrates were mixed with 0.4kg of CaCO3 and 10kg of rice bran. Pasteurization was achieved in sac bags using metal drums of about 500litres capacity with a wooden platform of about 35cm high within at 60-800C for 6 hrs (Fasidi, 2006). Initial water level in the drum was 30cm from the bottom of the drum. The pasteurized substrates were allowed to cool for 12 hrs, and then were divided into 3.5kg portions representing the content of each mushroom bag. Each of these portions was mixed with 2%, 4% or 6% of either GC or PKC separately before bagging. Bagging was done in an axenic condition. Plastic bags measuring 30x25cm were used. Each bag was inoculated with 5g of Pleurotus ostreatus. The open end of each bag was secured with a PVC pipe 3cm long and 3cm in diameter wrapped with a rubber band and plugged with cotton wool. 2.3 Spawn running The bags were hanged with ropes serially from the roof down with each line carrying a maximum of five bags. Spawn running room was allowed limited light. Room temperature was about 300C and relative humidity between 65 and 75% achieved by spraying the compost bags and walls of the spawn running room 2 to 3 times daily with clean water (Fasidi, 2006). 2.4 Cropping and harvesting At the end of spawn run, the bags of mycelial colonized substrates were transferred to the cropping room stacked on bamboo shelves and opened. This set up was sprayed with clean water after every 6hrs. The floor of the cropping room was covered with coarse sand from the sea bed and continuously moistened. The walls of the mushroom house were wetted regularly to humidify the environment to about 70%. On fruit bodies’ formation and maturity, data were taken on area of pileus, length of stipe using conventional 60
  • 3. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 methods. Fresh weight was taken using Agilent electronic balance. Data were analyzed using SPSS version 14.0. Biological efficiency (%) = Total weight of the fruit bodies___ X 100 Total weight of substrate (compost) 1 3. RESULTS 3.1 Spawn run Result from the study revealed that initiation of mycelial growth took place in both treated and untreated substrate samples within 7 days post inoculation (dpi). However, completion of spawn run was delayed in SD substrates treated to PKC beyond 2% and in BL substrates treated with 4% PKC and above and in BL+SD at all levels of PKC treatments. Spawn run in the BL, BL+SD and SD controls were completed in 21, 28 and 35 days respectively, following inoculation (data not shown). The uniformity in the initiation of mycelial growth in all the substrates may stem from the low hydrolase activity reported of P. ostreatus during substrate colonization (Elisashvili et al., 2008). The implication is that the level of breakdown of substrate materials for the release of nutrients is low or non-existent within this short period of 7 days, hence nutrients in the additives may have not been released and consequently not available to the mycelia and hence had no influence on the initial growth of the mycelia. The variation in the spawn run rate between the treated substrate samples may have resulted from the differences in the chemical composition of both the substrates and additives and the rate at which these materials were hydrolyzed. Guillen-Navarro et al. (1998) linked the production of mycelial biomass to the hydrolytic enzymes and availability of glucose in the substrate of cultivation. Variations in the speed of colonization (spawn run) by mycelia of different species of Pleurotus cultivated on different substrates supplemented with various additives have been reported (Stanley and Awi-Waadu, (2010), Renganathan et al., 2008, Pathmashini et al., 2008). However, the negative correlation observed between additive supplementation of substrates and the mycelial growth of P. ostreatus is in contrast with an earlier report (Renganathan et al., 2008) on impact of additives on the growth of mushrooms. The contrast noted may be a function of the type and availability as well as quantity- dependent interactions between the nutrient content of the additives and that of the substrates. Chemical content of palm kernel cake (PKC) revealed the presence low amounts of nutrients and particularly, proteins reported to be lowest in PKC among the seed cakes in Sub Saharan Africa (FAO, 1984). This may have contributed to the reduced growth performance on PKC supplemented substrates. 3.2 Cropping Primordia production in all the substrates took place within the first seven days of transfer of the bags to the cropping room, however, the earliest (4 days) was in the controls of the three substrates and the last (7 days) to produce primordial was in PKC- treated SD substrates. There were a total of 14 flushes within 10 weeks with a significant decline in growth parameters from the 8th week of harvest, especially in mushrooms cultivated on PKC-treated SD substrates. Assessment of the influence of substrates and additives on the pileus (cap) area of the test mushroom revealed that areas of mushroom cap were larger in untreated substrate samples compared with those produced on treated units (Table 1). The largest pileus area (40.64cm2) was recorded in mushrooms produced on BL substrates (control), while SD substrates (control) supported mushrooms with the least pileus area (24.82cm2). The disparity observed in the size of pileus produced on BL and SD substrates is likely a function of nutrient availability from the materials of the two substrates. Sawdust contains high amounts of lignin (a complex carbon source highly resistant to enzymatic degradation) than cellulose (a compound easily broken down to glucose which is the form taken in by the fungus as nutrient) and vice versa (Choi, 2004), hence glucose may have been readily available to the fungus grown on banana leaves substrates than those on sawdust. However, there were no significant difference (P<0.05) in pileus areas on SD substrates treated to either additives at all levels of concentration. Generally, addition of 4% of the additives on BL and BL+SD substrates produced mushrooms with smaller pileus size with the smallest 61
  • 4. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 (13.08cm2) observed on 4% PKC-treated BL substrates. This negative impact of additive on pileus size is probably consequent upon its influence on the mycelial growth (Baysal et al., 2006). When the performance of the additives were considered irrespective of the level of concentration, it was discovered that PKC supplemented sawdust substrates recorded a slightly significantly (P<0.05) larger pileus area than those grown on other substrates. This suggests that, concentration of PKC was the major determinant of the vigor of growth when added to sawdust substrates. BL and BL+SD substrates performed better when they were supplemented with GC+PKC. This result points to the fact that the presence of GC may have improved on the nutrient status of the substrate. Groundnut has been reported to be high in nutrients such as protein (25.2%), oil (4.8%) and 4.5% soluble sugars (Jambunathan, 1991). The additives did not have any significant effect on the length of stipe, irrespective of the substrate used except marginal differences observed in mushrooms produced on PKC supplemented SD substrates and in BL substrates supplemented with zero and 2% GC+PKC (Table 2 ). There were no significant differences observed in the length of stipe produced on the test substrates supplemented with different levels of concentrations of the two additives. However, the length of stipe produced on all the treated substrates were significantly (P<0.05) lower than those produced on untreated substrate samples (Table 4). This suggests that the growth and development of stipe was not influenced by the concentration levels of the additives, but by the very presence of the additives in the substrates. The influence of additives and substrates on the fresh weight of the test mushroom was assessed. A significant (P<0.05) decline in the fresh weight of P. ostreatus was observed when PKC was added to sawdust and when the same substrate was supplemented with GC+PKC beyond 2%. The decline in growth observed seems to be influenced by the introduction of PKC. The low protein status of this supplement may be the determining factor. FAO (1984) reported PKC to contain the lowest protein level among the oil seed cakes in Sub-Saharan Africa. On banana leaves substrates, the highest fresh weight of the test mushroom was recorded on the untreated samples. Treatment with additives did not encourage fresh weight accumulation in the mushroom cultivated on BL alone. However, when this substrate (BL) was supplemented with sawdust (BL+SD) and treated with PKC or GC+PKC, the mushroom recorded weight increases at all levels of the additives concentration (Table 5). The banana leaves sawdust composite (BL+SD) seems to be compatible with the additives when compared with sawdust or banana leaves alone. The reason for positive growth support compatibility of the additives with BL+SD substrates is likely resultant from the high cellulose content of banana leaves (which makes up for the highly lignolytic nature of sawdust) (Choi, 2004), and the high nutritional content of groundnut cake (FAO, 1984) which makes up for the low nutrient status of PKC. The difference between the mean fresh weight of mushroom produced on untreated (no additives) samples and those obtained from additive- supplemented substrates were in the range of 1.12 to 14.35g (Table 6). The weight differences were smaller in samples containing BL and GC but higher in sawdust-containing samples treated to PKC alone as the additives. The result obtained for fresh weight is as expected since the fruit body yield is largely dependent on the performance of the fungus at mycelial stage and primordia formation (Baysal et al., 2006). Irrespective of the concentration of the additives, the mean fresh weight of mushroom produced on sawdust treated to PKC (21.74g) and GC+PKC (22.21g) at the end of 10 weeks of harvest were not significantly different (P<0.05). However, the mean fresh weight produced on banana leaves substrate treated to GC+PKC was significantly (P<0.05) higher than the value obtained for samples treated with only PKC. Similar trend was observed in banana leaves + sawdust substrates (Table 7). The highest mean fresh weight (29.87g) of mushroom was recorded on banana leaves + sawdust substrates treated with GC+PKC while the lowest mushroom weight was seen on the sawdust substrate treated to GC+PKC. Biological efficiency (BE) of the three substrates treated with PKC and GC+PKC were assessed. Results showed that mushrooms grown on substrate samples without additives treatment recorded higher mean values of biological efficiency compared with values obtained on treated samples except for BL+SD substrates which recorded comparable BE values at all levels of additives treatment . Sawdust substrates treated with 2% GC+PKC and 6% PKC still recorded higher BE values. Generally, sawdust or banana leaves substrates supplemented with 4% and above of PKC were not efficient in the cultivation of P. ostreatus (Table 8). 62
  • 5. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 All growth parameters of Pleurotus ostreatus negatively correlated with PKC and GC + PKC supplemented sawdust substrates. Sawdust substrates were more biologically efficient when they were supplemented with 2% GC+PKC. Sawdust or banana leaves substrates supplemented with 4% PKC and above were not efficient in supporting the growth of P. ostreatus. Though, substrates were not supplemented with GC only, it is likely that supplementation of sawdust and banana leaves substrates alone or BL+ SD composite with GC alone may produce better results. Substrate supplementation with organic and/or inorganic substances often boosts mushroom production (Quimio et al., 1990, Renganathan et al., 2008, Adejoye and Mesewonrun, 2009). This is, however, true but with some reservations. The findings of this study revealed that substrate supplementation can only be result-oriented when the nutrient status of both the substrates and the supplement (additive) is considered complementary, and the additives’ concentrations adequate. This fact should be borne in mind by mushroom farmers, especially when PKC is the additive and sawdust or banana leaves the growth substrates. References Adejoye, O. D., Mesewonrun, O. (2009). The growth and yield of Lentinus squarrosulus (M.) Singer a Nigerian edible mushroom as affected by supplements. The Internet Journal of Nutrition and Wellness. Volume 8, Number 2. Baysal, E., Peker, H., Yalinkilic, M. K., Temiz, A. (2003). Cultivation of oyster mushroom on waste paper with some added supplementary materials. Bioresource Technology. 89(1):95-97. Chang, S. T., Miles, P. G. (1989). Edible mushroom and their cultivation. Florida.CRC Press. Choi, K. W. (2004). Shelf cultivation of oyster mushroom-with emphasis on fermentation. In Mushroom Growers’ Handbook. Mushworld. Pp. 153-165. Elisashvili, V., Kachlishvili, E., Penninckx, M. J. (2008). Lignolytic enzymes profile during growth and fruiting of Pleurotus ostreatus on wheat straw and tree leaves. Acta Microbiol. Immunol. Hung. 55(2): 157-168. Fasidi, I. O. (2006). Substrate sourcing and preparation for mushroom cultivation. Training workshop on cultivation of tropical mushrooms 20th -22nd November, 2006 at University of Ibadan, Oyo State. Guillen-Navarro, G. K., Marquez-Rocha, F. J., Sanchez-Vazquez, J. E. (1998). Production of biomass and lignlytic enzymes py Pleurotus ostreatus in submerged culture. Rev. Iberoam Micol. 15(4): 302- 306. Jambunathan, R. (1991). Groundnut quality characteristics. In Uses of tropical grain legumes: Proceedings of a Consultants Meet ing, 2 7 - 3 0 Mar 1989, ICRISAT Center, India. Patancheru. Pages 267- 275. A.P. 502 324, India: International Crops Research Institute for the Semi -Arid Tropics. Kong, W. (2004). Description of commercially important Pleurotus species. In Mushroom Growers’ Handbook. Mushworld. Pp. 55-61. Pathmashini, L., Arulnandhy, A., Wijeratnam ,R. S. W. (2008). Cultivation of oyster mushroom (Pleurotus ostreatus) on sawdust. Cey. J. Sci. 37(2): 177-182. Poppe, J. (2004). Agricultural wastes as substrates for oyster mushroom. In Mushroom Growers’ Handbook. Mushworld. Pp. 76-85. Quimio, T. H., Chang, S.T., Royse, D. J. (1990). Technical guidance for mushroom growing in the tropics. FAO publication. PP. 1-31. Renganathan, P., Eswaran, A., Balabaskar, P. (2008). Effect of various additives to the bed substrate on the sporophore production by Pleurotus flabellatus (Berk. And Br.) Sacc. Mysore J. Agric. Sci. 42(1): 132-134. Stanley, O. H., Awi-Waadu, G. D. (2010). Effects of substrates of spawn production on mycelial growth of oyster mushroom species. Res. J. Appl. Sci. 5(3)161-164. 63
  • 6. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Table 1: Influence of substrates and additives on area of pileus of Pleurotus ostreatus. Substrate (kg) /Additives (g) and Area of Pileus (cm2) SD BL BL+SD Concentration (g/kg) PKC GC+PKC PKC GC+PKC PKC GC+PKC 0 24.82 24.82 40.64 40.64 35.58 35.58 2 17.48 24.49 28.36 27.12 16.61 23.73 4 27.27 27.86 13.08 23.99 19.72 19.70 6 28.32 19.48 14.32 23.67 18.77 23.44 LSD 1.24 1.17 1.34 SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Table 2: Comparative influence of PKC and GC+PKC supplemented Sawdust, Banana leaves and Banana leaves + Sawdust substrates on the area of pileus of Pleurotus ostreatus. Substrate (kg) and Area of Pileus (cm2) Substrates SD BL BL+SD PKC 24.38* 22.78 22.13 GC + PKC 24.16 28.86* 25.61* t- Value 0.33 0.41 0.36 *Significantly different at P<0.05. SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. 64
  • 7. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Table 3: Influence of substrates and additives on length of stipe of Pleurotus ostreatus. Substrate (kg) /Additives (g) and Length of Stipe (cm) SD BL BL+SD PKC GC+PKC PKC GC+PKC PKC GC+PKC Concentration (g/kg) 0 (untreated) 2.66 2.66 3.33 3.33 3.27 3.27 2 2.10 2.74 1.84 3.30 2.45 2.66 4 2.00 2.01 1.73 2.38 2.53 2.72 6 2.00 1.89 2.41 1.97 2.44 2.29 LSD 1.21 1.03 1.15 SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Table 4: Comparative influence of PKC and GC+PKC supplemented Sawdust (SD), Banana leaves (BL) and Banana leaves + Sawdust (BL+SD) substrates on the length stipe of Pleurotus ostreatus. Substrate (kg) and Length of Stipe (cm) Substrate SD BL BL+SD PKC 2.19Ns 2.33 2.67Ns GC + PKC 2.33 2.75Ns 2.74 t- Value 0.21 0.43 0.11 SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Ns= no significant difference between treatments. 65
  • 8. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Table 5: Influence of substrates and additives on fresh weight of Pleurotus ostreatus. *Substrate (kg) /Additives (g) and Fresh weight (g) SD BL BL+SD PKC GC+PKC PKC GC+PKC PKC GC+PKC Concentration (g/kg) 0 27.87 27.87 33.75 33.75 31.17 31.17 2 13.91 30.69 15.80 24.12 30.34 35.12 4 21.08 16.67 19.92 15.86 37.38 21.30 6 15.74 13.59 12.46 26.69 25.30 31.89 LSD 1.36 *values are means of three determinations SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Table 6: difference in mean fresh weight of P. ostreatus produced on additive-treated and untreated mushroom substrates. Fresh weight (g) Substrates Untreated Treated Difference SD alone 27.87 - SD+PKC - 22.07 5.8 SD+GC+PKC - 22.25 5.62 BL alone 33.75 - BL+PKC - 19.40 14.35 BL+GC+PKC - 25.11 8.64 BL+SD alone 31.17 - BL+SD+PKC - 25.72 5.45 BL+SD+GC+PKC - 30.05 1.12 SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. 66
  • 9. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol 2, No.3, 2012 Table 7: Comparative influence of PKC and GC+PKC supplemented Sawdust (SD), Banana leaves (SD) and Banana leaves + Sawdust (BL+SD) substrates on the fresh weight of Pleurotus ostreatus. Substrate (kg) and Fresh weight (g) Substrate SD BL BL+SD PKC 19.65ns 20.48 31.03 GC + PKC 18.461 25.02* 29.87* t- Value 1.22 1.31 1.14 *significantly different at P<0.05; ns – not significantly different at P<0.05. SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. Table 8: Biological efficiency of Pleurotus ostreatus as influenced by different substrates and additives *Biological efficiency (%) SD BL BL+SD PKC GC+PKC PKC GC+PKC PKC GC+PKC Concentration (g/kg) 0 7.47 7.96 9.12 9.64 7.71 8.91 2 3.97 8.77 4.51 6.89 8.67 10.03 4 6.02 4.76 5.69 4.53 10.68 6.09 6 3.88 7.35 3.56 7.63 7.23 9.11 *Values are computed from means of 14 flushes. SD = sawdust, BL = banana leaves, PKC = palm kernel cake, GC = Groundnut cake. 67
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