CHAPTER II. world attention in recent years. The bioconversion of wastes to useful products has

Transcription

1 CHAPTER II Biochemical Analysis of Spent Mushroom Compost 2.1 Introduction The bioconversion of agriculture and industrial wastes into food has attracted the world attention in recent years. The bioconversion of wastes to useful products has tremendous potential in that it can help to meet the increasing world demand for food and energy. Likewise, many wastes like coir pith and paddy straw (Ramalingam et al., 2004), Green wastes from local vegetable market (Logakanthi et al., 2006) were decomposed by using mushrooms. The spent mushroom compost, which was prepared in the present study by using dry flower industrial waste, reduced the pollution caused by the waste of dry flower industry to some extent. Spent mushroom substrate is an excellent one to spread over the top of newly seeded lawns. The material provides cover against birds eating the seeds and will hold the water in the soil while the seeds germinate. The fresh mushroom compost applied to soil or incorporated into soil has many benefits: improves soil structure, provides plant nutrients, increases plant nutrient availability, increases soil microbial populations, increases soil cation exchange capacity increases plant root structures increases soil aeration, improves soil water status and reduces soil compaction (Courtney and Mullen, 2008; Mullen and McMahon, 2001; Maher et al., 2000) It improves the quality of compost by increasing high nutrient content. It is an attractive proposition for utilizing spent mushroom compost as soil inorganic fertilizer supplementation. The obtained composts were tested for the presence of various nutrients like C, Total N, S, H, Zn, Mg, Fe, Ni, Cu, Na, K and C/N ratio. Previous studies show that, the spent mushroom bed is an excellent source of Phosphorus, Potassium and other trace 63

2 elements (Mullen and McMahon, 2001; Kutuk et al., 1998; Stewart et al., 1998). The spent straw contains large quantity of N, P, K and can be used as manure (Maher, 1991). Pleurotus species were the rich source of proteins and minerals like Ca, P, Fe, K and Na and vitamin like C, B complex like thiamine, riboflavin, folic acid and niacin (Caglarmak, 2007; Patrabansh and Madan, 1997). They are consumed for their nutritive as well as medicinal values (Agrahar-Murugkar and Subbulakshmi, 2005). Mushroom protein is intermediate between that of animals and vegetables (Mamiro and Royse, 2008; Ndekya, 2002; Mshandete and Cuff, 2007) and is of superior quality because of the presence of all the essential amino acids (Purkayastha and Nayak, 1981). Pleurotus sp. contains high potassium to sodium ratio, which makes mushrooms an ideal food for patients suffering from hypertension and heart diseases. Ganoderma lucidum possesses the potential for anti-hiv activity (Chang and Mshigeni, 2001). The practice of mushroom cultivation not only produces a nutritious food but also improves the degradability of the straw. This takes place by reducing lignin, cellulose, hemi cellulose, tannin and crude fiber content of straw making it ideal for animal feed (Ortega et al., 1992). 2.2 Methodology Nutritional analysis: Analysis of moisture, protein, fat, crude fiber, total carbohydrates, ash of samples were done by standard methods (AOAC, 1995; Wankhede and Tharanthan, 1976). Zinc, Magnesium, Iron, Nickel, Copper, in samples were determined by using Atomic Absorption Spectrophotometry (Model: Elico SL-173 AAS). Sodium and Potassium were determined with Flame photometer (Model: Elico CL 361). Carbon, Nitrogen, Sulphur and Hydrogen were analyzed by Analytical functional testing Vario EL III CHNS (Serial No ). 64

3 2.3 Result and Discussion Mushroom cultured spent substrates are the mixtures of materials with high nutritional values including mushroom mycelia, degraded cellulosic fibers, degraded lignins, proteins, minerals, and so on which make them a recognized valuable biological resource particularly as compost (Kwak et al., 2008; Kim et al., 2007a,b) used the mushroom spent materials as animal feed. In this regard, the micro nutritional level assessed using the spent substrates of Ganoderma lucidum, Pleurotus sapidus, Pleurotus flabellatus as shown in Table 2.1, 2.2 and 2.3, the concentrations of the mineral components in the post harvest cultivation substrates are largely higher than those in the input cultivation substrates. This is probably due to the supply of mineral elements through moisture during the cultivation (Chang-yun Lee et al., 2009). Minerals in the diet of animals are essential for metabolic reactions, healthy bone formation, transmission of nerve impulses, regulation of water and salt balance (Kalac and Svoboda, 2000). The spent mushroom beds can be used as a fertilizer for plants. The nutritional component present in the organic wastes used in the study enhanced the growth of mushrooms. They also enhanced the mineral content of spent mushroom compost, which varied with different substrates and their combinations (Table ). Magnesium Magnesium (Mg) content of spent mushroom bed material of Ganoderma lucidum ranged from ppm. The highest value was observed in Trial III spent mushoom compost (15.86 ppm) which was followed by Trial II (15.34 ppm) and Trial IV (15.16 ppm). Magnesium content of spent mushroom bed material of Pleurotus sapidus ranged from ppm; while in Control it was ppm. In Pleurotus flabellatus it was ranged from ppm and in Control bed it was All these values observed were very 65

4 close to the Mg level (15.14 ppm) observed in the raw material (Table ). As for as the Mg level was concerned the result indicated that there were not much variations in all the three mushrooms or the preparations of different combinations of mushroom beds (Chang- Yun Lee et al., 2009; Ersin Polat et al., 2009). Iron Fe content of spent mushroom bed material of Ganoderma lucidum varied from ppm. The Trial III and II showed the presence of maximum Fe content of 3.78 and 3.65 ppm respectively. The Trial IV showed the presence of 3.88 ppm while, in Control it was about 2.94 ppm and in the raw material, it was about 4.09 ppm (Table 2.1). In case of the spent mushroom bed material of Pleurotus sapidus the Fe level showed no drastic changes in almost all Trials when compared to the raw material except in Trial I. In case of Trial I and in Control material the Iron content was almost same and it was about 3.86 and 3.96 ppm respectively (Table 2.2). In case of spent mushroom bed material of Pleurotus flabellatus the Fe content varied between ppm. The level of Fe decreased incase of Control material (3.83 ppm) and Trial I (3.94 ppm) while it was almost the same in case of Trial II, III and IV (4.14, 4.17 and 4.67 ppm) in relation to the raw material (Table 2.3) (Maniruzzaman Sikder et al., 2010; Chang-Yun Lee et al., 2009; Ersin Polat et al., 2009). Sodium The Sodium concentration of spent mushroom bed material varied sinificantly with different ratios of substrates used as well as different mushrooms cultured. The sodium concentration of Ganoderma lucidum in its spent mushroom compost had significantly increased in Trial I and Control ( ppm respectively) than in the raw material 66

5 (194.6 ppm). But it significantly decreased in an ascending order from Trial II, III and IV (168.65, and ppm) (Table 2.1). In Pleurotus sapidus spent mushroom compost the sodium concentration was more or less the same in case of raw material bed, Control and Trial I (194.6, and ppm respectively). The result indicated that, with regard to the presence of sodium, there was no change in the bed materials or in other words, the development of mushrooms on different Trials containing different ratio of raw materials could not alter the sodium level (Table 2.2). In Pleurotus flabellatus spent mushroom compost; also almost the same trend was observed. High sodium concentration was observed in Trial III ( ppm) and Trial IV (195.5 ppm). There was a significant drop in the level of sodium than the Control in Trial I and II ( and ppm) (Table 2.3) (Chang-Yun Lee et al., 2009; Danny Rinker et al., 2004). Potassium Potassium is commonly found in all cultivated and wild mushrooms (Mattila et al., 2001; La Guardia et al., 2005). In the present study also all the three cultivated mushrooms showed the presence of excellent quantity of potassium. In Ganoderma lucidum spent mushroom bed materials the quantity of K recorded was from ppm. The high quantity was recorded in Trial I ( ppm) followed by the Trial III ( ppm) Trial II ( ppm) and Trial IV ( ppm). The amount of K obtained in Control bed ( ppm) was very close to the Trial I. But in all the studied Trials the quantum of K observed had significantly increased than the K present in raw material. This indicated that the growth of Ganoderma lucidum enhanced the level of K in the bed material. More or less the same trend had developed in case of the bed materials of the Pleurotus sapidus and Pleurotus 67

6 flabellatus grown in various Trials (Table ) (Chang-Yun Lee et al., 2009; Ersin Polat et al., 2009; Danny Rinker et al., 2004). Nickel Ni content of spent mushroom bed materials of Ganoderma lucidum varied from ppm. The raw material showed the presence of 0.65 ppm of Ni content. In the Control bed material the amount of Ni present was only 0.42 ppm. In the present study, the cultivation of Ganoderma lucidum using the dry flower industrial wastes resulted in the significant depletion of Ni. In Trial I which included 25% of raw material, the Ni level reduced to 0.47 ppm, when the raw material level increased to 50%, 75% and 100% in Trial II, III and IV respectively the quantum of Ni present in bed material also increased accordingly (Table 2.1). More or less the same type of trend observed in case of the bed material in which the Pleurotus sapidus (Table 2.2) and Pleurotus flabellatus (Table 2.3) was cultured (Chang -Yun Lee et al., 2009; Danny Rinker et al., 2004). Zinc With reference to the heavy metal Zn, the spent mushroom bed material obtained after the culture of Ganoderma lucidum, Pleurotus sapidus and Pleurotus flabellatus showed very good result. The Zinc in the raw material was 1.05 ppm. While, in the Control material it was only 0.99 ppm. In case of Trial I and Trial II in all the three studied mushroom culture there was a significant drop in the level of Zinc was observed. It was about ppm in Ganoderma lucidum, 1.11 and 1.23 ppm in Pleurotus sapidus and 1.07 and 1.32 in case of Pleurotus flabellatus respectively. In case of Trial III and IV the level of Zinc showed an increased level, in almost all the three mushrooms studied (Table ) (Maniruzzaman et al., 2010; Chang-Yun Lee et al., 2009; Ersin Polat et al., 2009; Danny Rinker et al., 2004). 68

7 Copper Cu content of spent mushroom compost of Ganoderma lucidum varied from ppm. In the raw material the Copper content was estimated as 1.46 ppm, and in the Control bed material as 0.96 ppm, while, it gradually decreased in case of Trial I (0.97 ppm) and Trial II (0.99 ppm) where as it slightly increased incase of Trial III (1.32 ppm) and Trial IV (1.39 ppm) which contain 75 and 100% of the raw material (Table 2.1). More or less the same trend was observed in the spent materials of the Pleurotus sapidus and Pleurotus flabellatus (Table 2.2 and 2.3). This result indicated that the flurished growth of mushrooms on the dry flower materials absorbed remarkable quantum of heavy metals from the bed. Thus it reduces the toxic impact of these metals to certain extent. The studied heavy metals like Ni, Zn and Cu had decreased in all Trials than the raw material. Recent evidence has shown that mushrooms can absorb metal ions in high concentrations (Chang-Yun Lee et al., 2009; Ersin Polat et al., 2009; Bystrzejewska-Piotrowska et al., 2008; Gonen Tasdemir et al., 2008) and the metal absorption capability appeared to be species-specific (Alonso et al., 2003). Therefore, it is very natural for different mushrooms to exhibit a preferential difference in absorbing mineral components. It is notable that metal binding peptides, namely phytochelatins which are found in most eukaryotic cells and some prokaryotes, preferentially form complex with transition metals such as Zn, Cu, and Ni (Clemens, 2006). Therefore, the phytochelatins in the fruiting bodies can be one explanation to address the high uptake capacity of Zn, Cu, and Ni by the mushrooms. 69

8 The Ash content The ash content is an important indicative parameter for decomposition and mineralization of the substrate. The ash content was observed to have increased with the increase of saw dust and paddy straw ratio in the studied waste bed material. The ash content of spent mushroom compost of Ganoderma lucidum varied from %. The ash content of the raw material was very minimum and it was only 4.31% while in Control which contain the straw as a substrate the ash content was 23.8%. In Trial I, II, III and IV the ash content observed was about 22.17, 10.36, 8.39 and 4.19% respectively. The result indicated that as the addition of raw material percentage increased in the Trials I-IV, the quantum of ash formed had gradually decreased. More or less the same trend observed in case of the Pleurotus sapidus and Pleurotus flabellatus (Table ) (Meena Khwairakpam and Renu Bhargava, 2007; Singh et al., 2003; Atiyeh et al., 2000a; Kavian and Ghatnekar, 1991). Organic Carbon The organic carbon was found to have decreased from its initial higher value in all the Trials after composting. The reduction of organic carbon may be due to the growth of mushrooms, which resulted in and decomposition of waste (Mehalingam et al., 2008; Neena Dhiman and Battish, 2005; Chaudhari et al., 2000). The carbon content of raw material was 35.75%. From this value, the carbon content of Ganoderma lucidum spent mushroom content had drastically decreased in Trial I (25.64%) and Trial II (23.80%) while in case of the Control it was 25.86%. In case of Trial III and Trial IV, the level of carbon increased to and 32.97% respectively (Table 2.4). 70

9 In Pleurotus sapidus spent mushroom compost, the level of Carbon observed in Control was 27.97% while in the raw material it was 35.75%. In Trial I and Trial II the level of carbon decreased significantly (28.66 and 28.76% respectively) due to the decomposition activity of the mushrooms. But in case of Trial III and IV the level once again shot up to and 32.42% respectively because of the presence of huge quantum of raw material and less decomposition activity. Almost the same trend was observed in case of Pleurotus flabellatus culture also (Table 2.5 and 2.6) (Shahnawaz et al., 2009; Nageswaran et al., 2003). Nitrogen In the present study, the nitrogen content observed in the raw material was 0.65% and in Control 0.52%. The level of nitrogen had significantly increased in all Trials of Ganoderma lucidum spent material and it was 0.68, 0.74, 0.93 and 0.83 in case of Trial I, II, III and IV respectively (Swati Pattnaik and Vikram Reddy, 2010; Umamaheswari, 2005; Balamurugan et al., 1999; Kutuk et al., 1999; Rhoads and Olsan, 1995). Table 2.1 Various Mineral content (in ppm) in Ganoderma lucidum cultured spent bed materials. The values indicated are the mean of three observations and ± SD. Substrates Mg Fe Na K Ni Zn Cu Ash (%) Raw material 15.14± ± ± ± ± ± ± ±0.04 Control 15.4± ± ± ± ± ±0.21* 0.96± ±0.16 Trial I 13.27± ± ± ± ± ± ± ±0.08 Trial II 15.34± ± ± ± ± ± ± ±0.08 Trial III 15.86±1.81* 3.78± ± ± ± ± ± ±0.04 Trial IV 15.16±0.04* 3.88± ± ± ±0.05* 1.32± ± ±0.04 * indicated insignificant at 0.05% level. 71

10 Table 2.2 Various Mineral content (in ppm) in Pleurotus sapidus spent bed materials. The values indicated are the mean of three observations and ± SD. Substrates Mg Fe Na K Ni Zn Cu Ash (%) Raw material 15.14± ± ± ± ± ± ± ±0.04 Control 15.43± ± ±0.42* ± ± ±0.11* 0.72± ±0.21 Trial I 15.13±0.03* 3.86± ±0.03* ± ±0.25* 1.20± ± ±0.15 Trial II 15.28± ±0.06* ± ± ±0.06* 1.21± ± ±0.25 Trial III 15.22± ±0.05* ± ± ± ± ± ±0.02 Trial IV 14.86± ±0.03* ± ± ±0.07* 1.11± ±0.06* 4.16±0.03 * indicated insignificant at 0.05% level. Table 2.3 Various Mineral content (in ppm) in Pleurotus flabellatus spent bed materials. The values indicated are the mean of three observations and ± SD. Substrates Mg Fe Na K Ni Zn Cu Ash (%) Raw material 15.14± ± ± ± ± ± ± ±0.04 Control 14.87± ± ± ± ± ±0.12* 0.78± ±0.15 Trial I 14.34± ± ± ± ± ± ± ±0.03 Trial II 14.16± ±0.05* ± ± ± ± ± ±0.02 Trial III 14.83± ± ± ± ± ± ±0.08* 14.69±0.03 Trial IV 15.06± ± ± ± ± ±0.03* 1.44±0.12* 4.67±0.04 * indicated insignificant at 0.05% level. In Pleurotus sapidus and Pleurotus flabellatus spent mushroom compost also, the Nitrogen percentage range had significantly increased in almost all Trials. The increased percentage observed in all the Trials in the present study, might be due to the decomposition of raw materials because of the development of various mushrooms (Table ) (Shahnawaz et al., 2009). 72

11 C / N ratio The C/N ratio is a measure of quality of soil and its organic matter as an energy source. It is also an important determinant of humus types and is also related to soil moisture, ph and to many other soil properties (Lee, 1985). It is evident from the Tables 2.4 to 2.6, the spent material obtained from all the studied organisms, the C/N ratio gradually got decreased than the raw material (55.43). In the spent material of Ganoderma lucidum, the C/N ratio sharply fell from the C/N ratio of raw material level (55.43%) to and 32.07% in case of Trial I and II respectively while it increased in Trial III and IV (35.27 and respectively) since these two Trials included high quantity of raw materials. Almost the same trend was observed in case of Pleurotus sapidus and Pleurotus flabellatus spent materials (Table 2.4 to 2.6) (Shahnawaz et al., 2009). Protein In raw material, the estimated protein value was 4.03% and in Control, it was 3.28%. In spent material of Ganoderma lucidum the protein content increased gradually as the raw material content in the bed increased. The protein value significantly increased from 4.03 in raw material to 4.23% in Trial I, 4.65% in Trial II, 5.83% in Trial III and 5.20% in Trial IV. As in Ganoderma lucidum spent material the level of protein significantly increased in case of Pleurotus sapidus and Pleurotus flabellatus spent material also than the raw material (Table ). The increase in protein content of spent material in the present study may be because of decomposition of total carbohydrate, crude fiber, cellulose and hemi cellulose which were utilized by the mushroom from the stage of inoculation. The protein content of the spent mushroom compost may have increased because of the spread of mycelium in the substrate and secretion of extra cellular enzymes by the mushrooms (Shyam Sopanrao Patil et al., 2010). 73

12 Sulphur and Hydrogen The Sulphur and Hydrogen contents of the raw material were 1.58 and 3.66% respectively. But it steeply fell in Trial I and II of all the three mushroom species cultured spent mushroom compost material (Table 2.4, 2.5 and 2.6). The steep fall in the Sulphur and Hydrogen indicated the conversion of raw materials into the compost due to the utilization of bed materials by the fungus species (Hemalatha, 2012). And negative trend was observed in almost all fungus which were grown on the substrates which contain enormous quantity of the wastes (Trial III and IV which contain 75% and 100% raw material). Table 2.4 CHNS Analysis of Spent Mushroom Compost of Ganoderma lucidum. The values indicated are the mean of three observations and ± SD. Substrates S (%) H (%) C (%) N (%) C/N Ratio Protein (%) Raw material 1.58± ± ± ± ± ±0.02 Control 0.04± ± ± ± ± ±0.02 Trial I 0.92± ± ± ±0.05* 38.03± ±0.31* Trial II 0.33± ± ± ± ± ±0.22 Trial III 1.08± ± ± ± ± ±0.28 Trial IV 1.32± ± ± ± ± ±0.02 * insignificant at 0.05% level. Table 2.5 CHNS Analysis of Spent Mushroom Compost of Pleurotus sapidus. The values indicated are the mean of three observations and ± SD. Substrates S (%) H (%) C (%) N (%) C/N Ratio Protein (%) Raw material 1.58± ± ± ± ± ±0.02 Control 0.04± ± ± ± ± ±0.25 Trial I 0.14± ± ± ± ± ±0.05 Trial II 0.13± ± ± ± ± ±0.03 Trial III 1.07± ± ± ± ± ±0.34 Trial IV 1.13± ± ± ±0.10* 45.42± ±0.63* * insignificant at 0.05% level. 74

13 Table 2.6 CHNS Analysis of Spent Mushroom Compost of Pleurotus flabellatus. The values indicated are the mean of three observations and ± SD. Substrates S (%) H (%) C (%) N (%) C/N Ratio Protein (%) Raw material 1.58± ± ± ± ± ±0.02 Control 0.04± ± ± ± ± ±0.03 Trial I 0.11± ± ± ± ± ±0.29 Trial II 0.12± ± ± ± ± ±0.44 Trial III 1.29± ± ± ± ± ±0.44 Trial IV 1.36± ± ± ± ± ±0.35 * insignificant at 0.05% level. 2.4 Conclusion The biochemical analysis of spent mushroom materials obtained in the present study indicated that the nutrient content of the materials were enhanced to a greater extent after the rich growth of (Trial I and II) mushrooms. The total quantity of certain minerals and nutrients present in the raw material either reduced / increased, or tuned according to the requirement of the plants after the growth of mushrooms. This phenomena was well marked in almost all mushrooms cultured on Trial I and Trial II. Most of the minerals and nutrients did not reach the maximum required limit after the cultivation of mushroom. But one thing was definite that in all the cultured mushrooms, in all Trials the nutrient as well as the minerals level dropped to certain extent than in raw material. This itself may be considered as a very good result. 75