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Research Article - (2018) Volume 9, Issue 8

Fungal Incidence and Growth of Two Pleurotus Species on Sawdust of Ceiba pentandra (Linn.) Gaertn and Ficus Mucuso welw (Softwoods)

Sobowale AA*, Atoyebi FT and Adenipekun CO
Department of Botany, University of Ibadan, Ibadan, Nigeria
*Corresponding Author: Sobowale AA, Department of Botany, University of Ibadan, Ibadan, Nigeria, Tel: 234 8037265148 Email:

Abstract

Objective: Nigeria is faced with one of the most pressing environmental challenge which is the large production of sawdust waste of which Ceiba pentandra and Ficus mucuso are among the major contributors. This work seeks to harness waste through mushroom cultivation thereby reducing environmental hazards usually caused by improper disposal and burning. Softwood sawdust of Ceiba pentandra and Ficus mucuso was evaluated for their effects on the production of Pleurotus ostreatus and Pleurotus pulmonarius. Similarly, probable relationship among the fungal incidence of the substrates (sawdust) and that of mushrooms were examined.

Methods: The treatments used were replicated three times in a completely randomized design. The fruiting bodies were harvested and growth parameters, total yield and biological efficiency (BE) of the mushroom were recorded while their nutritional analysis was carried out. Resident fungi in the sawdust and mushrooms were isolated and identified after obtaining pure cultures. The data obtained were subjected to analysis.

Results: Ceiba pentandra and Ficus mucuso sawdust (substrates) supported the growth of the two mushrooms. P. ostreatus grew significantly (p ≤ 0.05) higher than that of P. pulmonarius. However, the substrates had significant (p ≤ 0.05) impact on the different growth parameters of the mushrooms. Generally, fermented sawdust significantly (p ≤ 0.05) improved some growth parameters of the mushrooms compared to unfermented one. Zero percent (0%) additive had significant (p ≤ 0.05) impact on growth parameters of the mushrooms than other concentrations. The mushrooms was nutritionally good. Similar fungi were isolated from the substrates (sawdust) and the mushrooms.

Conclusion: Softwood sawdust is thus a conducive substrate for mushroom cultivation and could be part of the solution to environmental challenge due to sawdust in Nigeria.

Keywords: Ferment; Resident fungi; Ceiba pentandra; Ficus mucuso; Pleurotus ostreatus; Pleurotus pulmonarius

Introduction

Mushrooms are one of man’s earliest form of fungi known [1]. They are macro-fungi, many of which are known to be edible [2]. Pleurotus species is an edible mushroom known to grow on any kind of lignocellulose waste (Chang and Milles 2004). Member of the species have received considerable attention due to their nutritional value, medicinal properties and biodegradation abilities [3]. Nigeria is known to produce high amount of sawdust waste among all other lignocellulose waste (softwood inclusive). Sawdust of softwood such as Ceiba pentandra and Ficus mucuso are known to be among the major contributors of this waste. Micro-fungi (Resident fungi) on the other hand are known to be among the common contaminants growing both on sawdust and mushrooms [4]. Mushrooms are widely distributed in nature with some being edible and some poisonous [1] Sawdust of certain trees such as mango and cashew have been reported to improve yield of different mushrooms [5,6]. Fermented and Unfermented sawdust have also been reported to affect the yield of different mushrooms [7]. Certain additive such as rice bran and wheat bran have also been reported to improve growth of mushroom [8]. However, different species of fungi have been isolated from sawdust [9]. Therefore, the experiment was set up to determine the effect of softwood sawdust in the production of Pleurotus ostreatus and Pleurotus pulmonarius and to also examine any probable relationship among the fungal incidence of the substrate (sawdust) and that of mushrooms.

Materials and Methods

Collection of sawdust and additive

The sawdust and additive were collected from sawmill in Isopako and the feed mill in Bodija market, Ibadan, Oyo State respectively. The sawdust was identified by Prof. A.A Jayeola in the Anatomy unit of Botany Department, University of Ibadan.

Collection and multiplication of spawn

Spawn was collected from the Plant Physiology Laboratory, Department of Botany, University of Ibadan. Multiplication of spawn was done using the method of Adenipekun and Fasidi [10]. The spawn bottles were stored in the refrigerator below 10°C.

Fermentation and preparation of the substrates

The fermentation of the substrates (sawdust) was carried out using the method of Gbolagade [11]. Preparation of fermented and unfermented substrates were done using the method of Adenipekun and Fasidi [10].

Inoculation and fructification of mushrooms

The bottles were inoculated with 10 g spawn of Pleurotus pulmonarius and Pleurotus ostreatus and were incubated at 28 ± 2°C for 3 weeks. They were later taken out and watered regularly for fructification.

Data collection

The fruiting bodies were harvested and the growth parameters, total yield and Biological Efficiency (BE) of the mushroom were recorded.

Isolation and identification of fungal species

The isolation of resident fungi in the sawdust and mushroom were done in the Plant pathology and mycology department. They were identified after obtaining pure cultures.

Determination of proximate composition (Analysis)

Proximate composition of Pleurotus ostreatus and Pleurotus pulmonarius was determined according to AOAC 2005.

Data analysis

The data obtained were subjected to analysis (ANOVA) using SAS (version 9.3). Means were separated (p ≤ 0.05) using Duncan’s Multiple Range Test (DMRT).

Results

Growth parameters of Pleurotus ostreatus and Pleurotus pulmonarius

Ceiba pentandra and Ficus mucuso sawdust supported the growth of the mushrooms and also had significant (p ≤ 0.05) impact on different growth parameters of the mushrooms. Generally, fermented sawdust significantly (p ≤ 0.05) improved some growth parameters of both Pleurotus ostreatus and Pleurotus pulmonarius compared to the unfermented one. Such parameters included; pileus thickness, number of fruiting bodies and biological efficiency. However, P. ostreatus was significantly (p ≤ 0.05) higher than that of P. pulmonarius in all the growth parameters. The mushroom’s first harvest was significantly higher than that of the second harvest (Table 1). Of all the wheat bran concentrations (additives), growth parameters of mushrooms that received 0% additive was significantly (p ≤ 0.05) higher than the growth in other concentrations (Table 2).

 Parameters Pileus length (cm) Pileus diameter (cm) Pileus thickness (cm) Stipe length (cm) Stipe width (cm) No of fruiting bodies Biological efficiency (%)
Mushroom species P. ostreatus 4.85a 5.68a 6.36a 7.16a 3.81a 5.02a 41.36a
P.pulmonarius 3.96b 3.96b 5.17b 4.48b 2.85b 3.09b 26.68b
Substrate types Ficusmucuso 4.26a 4.47b 5.53a 6.17a 3.38a 3.84a 32.82a
Ceibapentanda 4.55a 5.17a 6.01a 5.46b 3.29a 4.27a 35.22a
Substrate conditions Fermented 4.54a 4.63a 6.17a 5.64a 3.21a 4.60a 37.65a
Unfermented 4.27a 5.01a 5.36b 6.00a 3.45a 3.52b 30.39b
Harvest regimes First harvest 6.01a 6.87a 8.80a 8.30a 4.54a 6.24a 51.02a
Second harvest 2.79b 2.77b 2.74b 3.346b 2.12b 1.88b 17.02b
LSD0.05 0.48 0.57 0.57 0.64 0.39 0.45 2.91
R2 0.53 0.59 0.73 0.64 0.5 0.73 0.83

Means with different letters within the same columns are significantly different at p ≤0.05

Table 1: Effect of sawdust and harvest regime on the growth parameters of Pleurotus ostreatus and Pleurotus pulmonarius.

Parameters Pileus length (cm) Pileus diameter (cm) Pileus thickness
(cm)
Stipe length (cm) Stipe width (cm) No of fruiting bodies Biological efficiency (%)
Wheat bran concentrations 0 4.13a 5.36a 6.14a 5.69ab 3.44ab 5.04a 51.78a
10 4.50a 4.57ab 5.78ab 5.96ab 3.27ab 3.85b 33.24b
20 4.79a 5.12a 6.09a 6.44a 3.68a 3.77b 27.79c
30 4.19a 4.24b 5.05a 5.18b 2.93b 3.56a 23.26d
LSD0.05 0.67 0.81 0.81 0.9 0.56 0.64 4.12
R2 0.53 0.59 0.73 0.64 0.5 0.73 0.83

Means with different letters within the same column are significantly different at p ≤0.05

Table 2: Effect of additive on growth parameters of Pleurotus ostreatus and Pleurotus pulmonarius.

Fungal incidence in the mushroom species and sawdust

All the resident fungi isolated from the substrate (sawdust) were also isolated from the mushrooms (Table 3). The isolated fungi include Aspergillus niger, A. flavus, A. tamarii, A. fumigatus, Trichoderma harzinum, Trichoderma sp. 1, T. viride and Trichoderma sp. 2 (Figures 1-3).

plant-pathology-microbiology-fruiting

Figure 1: Aspergillus niger obtained from the substrates and fruiting bodies of Pleurotus ostreatus and Pleurotus pulmonarius (a) with its photomicrograph (b).

plant-pathology-microbiology-substrates

Figure 2: Aspergillus flavus obtained from the substrates and fruiting bodies of Pleurotus ostreatus and Pleurotus pulmonarius (a) with its photomicrograph (b).

plant-pathology-microbiology-photomicrograph

Figure 3: Trichoderma harzianum obtained from the substrates and fruiting bodies of Pleurotus ostreatus and Pleurotus pulmonarius (a) with its photomicrograph (b).

Parameters Variable Aspergillusniger A.flavus A. tamari A.fumigatus T.harzinum Trichoderma sp. 1 Trichoderma sp. 2
Mushroom Species P. ostreatus 0.33a 0.31a 0.21a 0.17a 0.25a 0.17 a 0.13a
P.sajorcaju 0.35a 0.31a 0.13a 0.15a 0.29a 0.04b 0.13a
Substrate types F. mucuso 0.35a 0.35a 0.17a 0.15a 0.27a 0.15a 0.15a
C.pentandra 0.33a 0.27a 0.17a 0.17a 0.27a 0.06a 0.10a
Substrate conditions Fermented 0.33a 0.33a 0.10a 0.13a 0.33a 0.04b 0.15a
Unfermented 0.35a 0.29a 0.23a 0.19a 0.21a 0.17a 0.10a
LSD0.05 0.13 0.18 0.15 0.15 0.18 0.12 0.13
R2 0.57 0.14 0.15 0.02 0.12 0.13 0.10

Means with different letters within the same column are significantly different at p ≤ 0.05.

Table 3: Fungal incidence in the mushroom species and the two sawdust.

Overall incidences of the fungi in the two mushrooms and sawdust

The overall incidence of the fungi (p ≤ 0.05) in mushroom species and sawdust were not significantly different from each other (Table 4). However, the comparison of the overall incidences of the fungi in the two mushrooms and sawdust were significantly different from each other (Table 5).

Parameters Variables Means
Mushroom species P. ostreatus 0.22a
P. pulmonarius 0.20a
Substrate types F. mucuso 0.23a
C. pentandra 0.20a
Substrate conditions Fermented 0.20a
Unfermented 0.22a
LSD0.05 0.06
Wheat bran concentrations 0 0.24a
10 0.22a
20 0.17a
30 0.21a
LSD0.05 0.09
R2 0.06

Means with different letters are significantly different at p ≤ 0.05

Table 4: Overall incidences of the fungi in the two mushrooms and sawdust.

Parameter Variables Means
    Pooled fungi     A. niger 0.34a
A. flavus 0.31a
A. tamari 0.17bc
A. fumigatus 0.16c
T. harzianum 0.27ba
Trichoderma. sp. 1 0.10c
Trichoderma. sp. 2 0.13c
LSD0.05 0.11
R2 0.06

Means with different letter are significantly different at p 0.05

Table 5: Comparison of the overall incidences of the fungi in the two mushrooms and sawdust.

Proximate composition of the mushroom species grown on the two sawdust

The two selected mushrooms grown on Ceiba pentandra and Ficus mucus sawdust were rich in protein, fibre, ash, and carbohydrate. However, the additives had different impact on the nutritional composition of the mushrooms (Table 6).

Parameters Moisture content Crude protein Crude fat Crude fibre Ash CHO
Mushroom species Pleurotus ostreatus 15.32a 32.15b 1.10a 23.30a 9.90b 38.20a
Pleurotus pulmonarius 16.27a 45.93a 0.92b 19.61b 11.40a 29.26b
Substrate condition Unfermented 15.61a 40.19a 1.05a 22.55a 9.83b 35.63a
Fermented 15.79a 37.90b 0.96b 20.36b 11.47a 31.83b
Substrate types Ceiba pentandra 16.33a 39.16a 1.08a 18.96b 10.89a 32.83a
Ficus mucuso 15.07a 38.92a 0.93b 23.95a 10.40b 34.63a
LSD0.05 2.52 1.41 0.08 1.88 0.35 3.31
Wheat bran concentrations 0 13.00b 32.97c 0.79c 23.46a 9.48c 44.61a
10 23.54a 43.14a 1.16a 17.12c 10.75b 26.52c
20 11.51b 40.23b 1.08ab 20.53b 10.86b 37.01b
30 14.75b 39.82a 1.00b 24.70a 11.50a 26.78c
LSD0.05 3.57 2 0.12 2.66 0.5 4.68
R2 0.4 0.96 0.49 0.37 0.69 0.56

Means with different letters in the same column are significantly different at p ≤ 0.05

Table 6: Proximate composition of mushroom species grown on the two sawdust.

Effect of fermentation pH and temperature on the two sawdust during fermentation

After fermenting the two sawdust for twelve days, the temperature and pH between the two substrates were significantly different (Figures 3-5).

plant-pathology-microbiology-fermentation

Figure 4:Effect of pH on the two-saw dust during fermentation.

plant-pathology-microbiology-temperature

Figure 5:Effect of temperature on the two saw dust during fermentation.

Discussion

The significant effect of substrates and additives on the growth parameter of P. ostreatus and P. pulmonarius confirmed the appropriateness of the model used. The enhanced growth of P. ostreatus and P. pulmonarius recorded in C. pentandra and F. mucuso sawdust agreed with the findings of Kadiri and Fasidi [12]; Fuwape et al. [13] which reported that lignocellulose wastes are good growth media for Pleurotus mushrooms. The variations observed in the growth parameters of the mushrooms species might be attributed to the impact of ecological factors, texture, water holding capacity, degree of aeration formulations and nutrients in substrates possibly affected the final mushroom growth [14-16]. However, the different effect of the two sawdust on some the mushrooms’ growth parameters corroborated the findings in the study of Ahmed [17] who reported significant influence of various substrates on the growth parameters. The increased biological efficiency of P. ostreatus and P. pulmonarius at the different substrate conditions indicated might be as a result of the fermented substrates as found in line with the reports of Hernandez et al. [18] who also recorded the highest biological efficiency in the fermented substrate. However, the better result obtained at 0% additive compared to other concentrations levels was in agreement with Soniya et al. [19] who revealed that the yield of rice straw without supplementation was the most significant highest among the treatments. The significant harvest rate recorded in the first harvest over the second harvest corroborated the report of Soniya et al. [19] who reported that the yield of oyster mushroom recorded the highest significance in the first flush of all treatments. This could possibly be a result of P. ostreatus and P. pulmonarius utilization of the growth nutrient mostly during the first harvest, thus resulting in declining in mushroom yield in the second harvest [20,21]. The eight resident fungi isolated from the substrates and mushrooms are in agreement with the findings of Lennox et al., Obire and Amadi [22] that also isolated Penicillium sp., Mucor sp., Trichoderma sp. and Aspergillus sp. from fermented sawdust. One of the fungi major characteristic is the ability to help in sawdust degradation according to the report of Obire and Amadi [22]. The similarity in the fungal incidence of the sawdust and mushroom species can be as a result of the interaction between them. The nonsignificant effect of the overall incidence of the fungi in the mushroom species and sawdust shows that the incidence of each fungus does not have any effect on the mushroom species and sawdust. However, the significance in the comparison of the overall incidences of the fungi in the two mushrooms and sawdust shows that incidence of each fungus occurred independently. This implies that when exposed to suitable conditions it can be virulent [23]. However, in spite of this, fungi produces lignocellulosic enzymes that help in the fermentation of the substrates because the medium composition, quality aeration rate, pH and temperature of the substrates used in mushrooms cultivation have an effect on the growth, chemical, functional and sensorial characteristics of mushrooms [24,25]. The pH of the substrates ranged from 6.90 - 7.00 during the fermentation process which agrees with the work of Urben [26]; Kalmis et al. [27] which reported the optimal pH range for the development of mushroom as 4.0-7.0. Fermentation is therefore necessary to adjust the pH level of the substrate pH [25]. The variation in the fermentation temperature could be due to the changes in the environmental conditions while the gradual increase in the temperature contrasted the findings of Sher et al. [28]. who reported that lower temperatures and dry condition can cause a reduction in the stalk height and cap size of mushroom. The richness of the two mushrooms supported the earlier findings that mushrooms have nutritional attributes and can be used as supplements [29,30]. Furthermore, the moisture content of mushroom depend on their harvesting time, maturation period and environmental conditions such as humidity and temperature in growing period and storage conditions [31]. The relatively low moisture content obtained in this study is in line with the report of low moisture content in some mushrooms collected in Nigeria and this indicated that mushrooms can be easily dried and have an extended shelf life [32,33]. The high crude protein and total carbohydrate content obtained in this study corroborates the works of Kadiri and Fasidi [12]; Kayode et al. [32]; Adejumo and Awosanya [34]. This indicates that mushrooms can replace animal protein and can be used as supplement in cereals or carbohydrate meals because of its affordability, medicinal and nutritive attributes. The crude fat content of the mushrooms was low and this agrees with the findings of Adebiyi et al. [35] and contracts the works of Akindahunsi and Oyetayo [36]; Khurdishul [37]. The relatively low fat content of the mushrooms showed that they have no cholesterol, suitable as a component of weight restricted diet and are less harmful to health than the saturated fatty acids [38]. The high crude fiber content obtained contrasts the work of Kayode et al. [32]. However, high fiber content in mushrooms indicated that it is a good source of roughages, thus having ability to aid easily passage of feaces from the body, reducing the risk of been exposed to several diseases [35]. The high ash content obtained in this study could be based on the role of ash content in the determination and characterization of the content of mineral substances which implies that such mushroom is a good source of minerals [39]. The substrates, substrate conditions, wheat bran concentration and mushroom species significantly (p ≤ 0.05) affected moisture content, crude protein content, crude fiber content, total ash content and carbohydrate contents of the selected mushrooms. The notable variation in the proximate composition of the mushrooms depends on the type of mushroom, the stage of development, the part samples, level of nitrogen available, the environments, storage conditions, additives and the substrate [40,41].

Conclusion

Mushroom cultivation on sawdust is an efficient way of recycling and managing agricultural and industrial waste. Mushroom farmers are encouraged to use softwood sawdust because they are easily accessible with no cost implication and also support mushroom growth. The mushrooms, which are relatively cheap and easy to come by, were rich in nutrients and therefore should be incorporated into our daily diets to improve its quality, nutritional and medicinal benefits. Further studies, should be done on the substrates composition before addition of additives. It is also important for the environmental conditions of the mushroom house to be kept sterile to limit the fungal incidence on mushroom.

Funding Information

No funding was received for this research.

Compliance with Ethical Standards

Conflicts of interest

All authors declare that there is no conflict of interest.

Ethical approval

This work does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual involved participants included in the study.

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Citation: Sobowale AA, Atoyebi FT, Adenipekun CO (2018) Fungal Incidence and Growth of Two Pleurotus Species on Sawdust of Ceiba pentandra (Linn.) Gaertn and Ficus Mucuso welw (Softwoods). J Plant Pathol Microbiol 9: 448.

Copyright: © 2018 Sobowale AA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.