Oyster Pleurotus ostreatus Mushroom Growth Yield from Probiotic Water Supplement PDF

OYSTER Pleurotus ostreatus MUSHROOM GROWTH YIELD FROM PROBIOTIC WATER SUPPLEMENT LIFE RESSEARCH IN SIP TEAM PRESENTED TO DIVISION OF SULTAN KUDARAT IRENE NERI CASSANDRA NICOLE PAÑADA THERESE REYES...

OYSTER Pleurotus ostreatus MUSHROOM GROWTH YIELD FROM PROBIOTIC WATER SUPPLEMENT LIFE RESSEARCH IN SIP TEAM PRESENTED TO DIVISION OF SULTAN KUDARAT IRENE NERI CASSANDRA NICOLE PAÑADA THERESE REYES RESEARCHERS JULY 2023  TABLE OF CONTENTS PRELIMINARY PAGES Page Title Page 1 Table of Contents 2 Lists of Tables 3 Lists of Figures 3 Abstract 4 INTRODUCTION Introduction 6 Statement of the Problem 9 Scope and Delimitation 11 Significance of the Study 11 MATERIALS AND METHODS Research Design 14 Research Locale 15 Research Procedure 16 Tissue Culture Spawn Production (DA Adaptation) 17 a. Tissue Culture Mushroom 21 b. Spawn Preparation 22 c. Inoculation of Spawn 24 Oyster Mushroom Cultivation (Researchers Procedure) 26 a. Substrate Preparation 26 b. Spawning/Planting Procedures 28 c. Spraying Procedures 30 d. Harvesting and Weighing 32 RESULTS AND DISCUSSIONS 35 Conclusion 41 Recommendations 42 BIBLIOGRAPHY 43 ACKNOWLEDGEMENTS 45 2  LISTS OF TABLES Table Title Page Flower Production of Oyster mushroom Pluerotus 1 osteatus on different fruiting bags treated with varying 35 water treatments. Analysis of Variance for the treatment Means on weight 2 37 level of flowers among fruiting bags treated with different water mixtures on Oyster mushroom Pleurutos ostreatus LISTS OF FIGURES Figure Title Page Conceptual Framework of the Study for Oyster 1 Mushroom Cultivation Exposed to Different Water 14 Mixtures Map of the Research Locale 2 15 Flow Chart on the Oyster Mushroom Cultivation in 3 Fruiting Bags Exposed to Different Water Mixtures 16 Comparison of Treatment Means on the growth 4 38 performance (weight) level of Oyster Mushroom 3  ABSTRACT NERI, IRENE, PANADA, CASSANDRA NICOLE, REYES, THERESE. “OYSTER Pleurotus ostreatus MUSHROOM GROWTH YIELD FROM PROBIOTIC WATER SUPPLEMENT” Research Capstone Project, New Panay National High School, New Panay, Esperanza, Sultan Kudarat. This study evaluated the effectiveness of different water mixtures as fertilizer supplements for promoting the growth of Oyster mushrooms (Pleurotus ostreatus). Conducted at the New Panay National High School Mushroom Center in New Panay, Esperanza, Sultan Kudarat, the research implemented an Input-Process-Output framework with five treatments: control (water), rice wash, water and salt, water and sugar, and water and probiotic milk. The study focused on assessing the number of flowers and weight of mushrooms as growth parameters. Treatment 4, utilizing water and probiotic milk, demonstrated the highest number of flowers, indicating its effectiveness in enhancing flower production. Furthermore, Treatment 4 resulted in the heaviest weight of mushrooms, highlighting its superior performance in overall growth and weight gain. Statistical analysis indicated no significant difference in flower weight among the fruiting bags treated with different water mixtures, suggesting that the choice of water mixture did not significantly influence flower weight. Based on these findings, Treatment 4 (water and probiotic milk) emerged as the most effective nutrient enhancer for cultivating Oyster mushrooms, warranting its prioritization as a fertilizer supplement due to its positive impact on flower production and weight. Future research should focus on elucidating the underlying mechanisms responsible for the growth-promoting effects of water and probiotic 4 milk, optimizing application methods and dosages to maximize benefits in Oyster mushroom cultivation. Keywords: Oyster mushroom cultivation, Nutrient enhancer, Probiotic milk, STEM Research Capstone 5  INTRODUCTION Background of the Study Mushrooms are fungi with significant nutritional value currently counting around 2000 edible species distributed around the world (Rathore et al., 2019). The most cultivated species include button mushroom (Agaricus bisporus), shiitake mushroom (Lentinula edodes), and oyster mushrooms (Pleurotus spp.). The current global market value of fresh mushroom reached 38 billion US dollars in 2018, and China is the largest mushroom producer within the Asian region, contributing approximately 35 % to the global mushroom market (Elaine and Tan, 2009). Asia countries contribute up to 76 % of mushroom production, followed by Europe (17.2 %) and United States (5.9 %) (Sande et al., 2019). Oyster mushroom (Pleurotus spp.) is one of the most cultivated mushroom species globally, which possess high demand and many beneficial properties (e.g., pharmaceutical properties). Therefore, it would be useful to review and discuss the different techniques of cultivation and valorization, and the factors affecting the growth and yield of oyster mushroom in mushroom cultivation industry. The use of various valorization techniques aiming at mitigating hazards and recovering value-added products from WMS is also presented and reviewed to provide an overview and key insights of the current progress in disposal and valorization of this abundant waste 6 generated in mushroom cultivation industry. Error: Reference source not found Sharma et al. (2013) reported that cultivation of oyster mushroom (Pleurotus ostreatus) has increased tremendously throughout the world because of their abilities to grow at a wide range of temperature and utilizing various agro-based residues. Pleurotus species are efficient lignin degraders, which can grow on different agricultural wastes with broad adaptability to varied agro-climatic conditions. Mushrooms with their flavor, texture, nutritional value and high productivity per unit area have been identified as an excellent food source to alleviate malnutrition in developing countries. P. osteratus are rich source of proteins, minerals & vitamins. Apart from food value, its medicinal value for diabetics and in cancer therapy has been emphasized. The practice of mushroom cultivation not only produces medicinal and nutritious food but also improves the straw quality. This takes place by reducing lignin, cellulose, hemicelluloses, tannin and crude fiber content of straw making it ideal for animal feed.Error: Reference source not found Consistent with the claims of Stamets (2000) and Lindequist et al., (2005), edible mushrooms are nutritionally endue fungi (mostly Basidiomycetes) that grow naturally on the trunks, leaves and roots of trees also as decaying woody materials. These edible mushrooms include Agaricus spp. (button mushrooms), straw mushroom (oil palm 7 mushrooms), Jew's-ear (wood ear mushroom), also as oyster mushroom (oyster mushrooms). According to Raboy (2021) rice wash’s use as a booster and 50% RS + 50% SD as substrates are recommended for mushroom production. Different proportions of the substrates and boosters evaluated must also be tried. Also, other available indigenous substrates and possible boosters in the locality should be explored. Analyses of the inorganic ions confirmed the existence of a characteristic chemical composition of the many monocot tyledonous salt marsh plants therein they contain high levels of potassium and comparatively low levels of sodium. Sugar is a carbohydrate. It is sweet, colorless, water-soluble compounds present in the plants and also in the milk of mammals. Sucrose is found in most plants, but it occurs at concentrations high enough for economic recovery only in sugarcane (Saccharum officinarum) and sugar beets (Beta vulgaris). Probiotic sour cream dip was developed using Lactobacillus casei strain Shirota from yakult as starter adjunct. Chemical composition (moisture, fat, protein, ash and total solid contents and pH) as well as putative lactic acid bacterial (LAB) counts were determined. Thus, the researchers ought to prove in an experimental study that different water mixtures as supplement is an effective type of watering method for the growth of Oyster Pluerotus ostreatus mushroom. 8 Statement of the Problem This study aimed to determine the effectiveness of different water mixtures as fertilizer supplement to influence the growth performance of Oyster mushroom Pluerotus ostreatus. Thus, the following questions are formulated. 1. What is the fruit production growth performance of oyster mushrooms Pluerotus ostreatus exposed in varying water mixtures in terms of: a. Rice wash, b. Water and salt, c. Water and sugar, and d. Water and probiotic milk? 2. What is the weight production growth performance of oyster mushroom Pleurotus ostreatus exposed in varying water mixtures of: a. Rice wash, b. Water and salt, c. Water and sugar, and d. Water and probiotic milk? 3. Is there a significant difference in the number of fruit and growth performance of Oyster Pluerotus ostreatus mushroom 9  treated with four different water mixtures as fertilizer supplement? 4. Which among the different water mixtures as fertilizer supplement is more effective in growing Oyster Pluerotus ostreatus mushroom? 10  Scope and Delimitation The experimental setup on fruiting bag was limited to sawdust, agricultural lime, rice bran (tiki-tiki), and sugar mixture that serve as our substrate and rice wash, water and salt, water and sugar, and water and probiotic milk as our fertilizer supplement for the production of Oyster Pluerotus ostreatus mushroom. The study was conducted at the School Oyster Mushroom House on November 2022. Sawdust was collected throughout the community and at the researchers' home. There were eight (8) fruiting bags of sawdust prepared each treatments. Each treatment bottles consists of Treatment 1- water (control), Treatment 2- water and salt, Treatment 3- water and sugar, and Treatment 4- probiotic milk and water. Significance of the Study The significance of the study on the growth performance of Oyster mushroom using different water mixtures as fertilizer supplements is multi-faceted and can benefit various individuals and groups: The findings of the study can be valuable for farmers who are looking to diversify their income sources. By demonstrating the 11 effectiveness of using sawdust mixture and different fertilizer supplements for cultivating Oyster mushrooms, farmers can consider integrating mushroom cultivation alongside their existing rice crops. This can potentially lead to increased profitability and economic stability for farmers. The study's results can assist the Department of Agriculture in developing strategies for mushroom production. By understanding the optimal water mixtures and fertilizer supplements for Oyster mushroom growth, the department can promote and support mushroom cultivation on a larger scale. This can contribute to ensuring sufficient mushroom supply and improving the agricultural sector's economic viability. The study's outcomes can have a positive impact on the entire community. By encouraging the utilization of sawdust and promoting mushroom cultivation as a business opportunity, the community can benefit from the creation of new economic ventures. This can lead to job creation, increased income, and overall community development. The study can be beneficial for academic institutions by providing students with exposure to scientific research and the proper procedures of a science investigatory project. It enhances the students' abilities to conduct scientific studies, encourages critical thinking, and fosters a deeper understanding of agricultural practices and their impact on food production; and 12  This research offers student researchers the opportunity to develop their skills in conducting scientific studies. By engaging in the research process and generating new knowledge, students can enhance their understanding of the subject matter and gain practical experience in real-life situations. This can foster a passion for scientific inquiry and inspire future careers in research or related fields. 13  METHODOLOGY Research Design This study was guided by experimental research design arranged Input-Process-Output Design using 5 different water mixtures as nutrient enhancer to wit: Control- Water, T1- Rice wash, T2- Water and Salt, T3- Water and Sugar, T4-Water and Probiotic Milk as it is sprayed in fruiting bags 2 times a day. INPUT PROCESS OUTPUT Preparation/s of water Treatment and Effective growth mixtures for Oyster observation of Oyster production of Oyster Pluerotus ostreatus Pluerotus ostreatus Pluerotus ostreatus mushroom substrate as mushroom growth mushroom in terms of fertilizer supplement under treatments of: performance under fruit and weight varying water mixtures Control- Water as fertilizer supplement T 1- Rice wash T2- Water & Salt T3- Water & Sugar T4- Water & Probiotic Milk Figure 1. Conceptual Framework of the Study for Oyster Mushroom Cultivation Exposed to Different Water Mixtures The conceptual framework focuses on studying the effects of different water mixtures as fertilizer supplements on the growth performance of Oyster Pluerotus ostreatus mushrooms. The study involves four treatments: a control group with plain water, rice wash, 14 water and salt, water and sugar, and water and probiotic milk. The water mixtures are prepared and used as supplements for mushroom substrate. The mushrooms are grown under controlled conditions, and their growth is observed in terms of fruit production and weight. The goal is to determine which water mixture, if any, results in improved mushroom growth. Research Locale The study was conducted at New Panay National High School Mushroom Center at New Panay, Esperanza, Sultan Kudarat beginning on the November 2022. All tests results were observed and recorded for statistical procedure. Figure 2. Map of Research Locale 15  Research Procedure Figure 3. Flow Chart on the Oyster Mushroom Cultivation in Fruiting Bags Exposed to Different Water Mixtures Preparation of Materials Materials used in this study were borrowed at the Science Laboratory of New Panay National High School. Materials needed are: alcohol lamp, spoon, beaker, and weighing scale. The spoon had undergone the sterilization process. Working area is free from any microorganism. a. Preparation of Sawdust Substrate The rice straw was cut into tiny bits, fully soaked in water for 48 hours and air dried for 24 hours. About 15 kilograms of rice straw substrate was weighed using weighing scale. The 500 grams of rice straw bits was put into plastic bags. b. Spawning Procedures A half tablespoon of Sorghum seed was put in the 500 grams of rice straws’ fruiting bags. Rubber bands and small amount of rounded 16  cotton was used to tie the fruiting bags to avoid bacteria that can contaminate sorghum inside the fruiting bags. c. Spraying Procedures Four different water mixtures as supplement were used to determine the most effective water treatments for the growth of oyster mushroom. The treatments were applied once a day only. d. Harvesting and Weighing Procedures The Oyster mushroom was harvested from different fruiting bags using a cutter or a hand only and weighed in a weighing scale. Tissue Culture Mushroom Spawn Production and Preparation of materials (adopted from Department of Agriculture training kits) Potato Dextrose Agar Preparation Ingredients: In order to prepare Potato Dextrose Agar, the researcher employed a variety of ingredients, including 300 grams of potatoes, 18 grams of agar (gulaman), 18 grams of white sugar, and 1liter. distilled water Materials: The researcher used a funnel to transport liquids, storage bottles for mixtures, and water that had been cooked in a kettle to make potato dextrose agar. To prevent contamination, cotton plugs are used to seal holes or remove impurities from liquids. To separate 17 or strain liquids, use cheesecloths and strainers. Cutting boards give you a level surface on which to cut with a knife. Using a weighing scale and a stirring rod to blend ingredients, precise measurements are taken before the concoction is wrapped in aluminium foil. Last but not least, a gas burner provides the heat for cooking. A pressure cooker, also known as an autoclave, uses high pressure to quickly and efficiently prepare food. Procedures: 1. Peel (250 g) potato and cut into cubes 2. Cook potato in 1 liter of distilled water 3. Separate potato broth by passing in a clean cheese cloth/ strainer 4. Add distilled water to the broth to make it 1 liter. Heat the broth in a low fire and mix agar and sugar. Stir continuously until the agar and sugar melts. 5. Dispense desired amount in clean bottle(s). 18 6. Cover bottle(s) with cotton plugs and aluminum foil. Slant the bottles and let the PDA solution cool and solidify. 19  A. Tissue Culture Mushroom Materials: In tissue culture mushroom to detect the presence of yeasts and molds in product samples, researchers need PDA solution; disinfectant to kill germs on surfaces and objects; an isolation chamber for maximum protection and operational safety; a scalpel or blade for making skin incisions and dissecting tissue; tissue paper to filter out excess oil; fresh mushrooms to cultivate as the primary source of growth; petri plates to hold growth medium in which cells can be cultured; and, finally, an alcohol lamp for heating, sterilizing, and burning in a lab. Procedure: 1. Wash the sorghum until the water is clear. 2. Cut the mushroom stem 3. Get a small amount of tissue in the center of the cut mushroom and put it in the center on the PDA solution (plated petri dish or bottle slant) 21 4. Seal properly the petri plate with the mushroom tissue, invert the plate and put it on a dark room for incubation. 21  B.Spawn Preparation Materials: In order to prepare spawn, researchers need to grow pure culture mushrooms, produce alcoholic beverages using sorghum seeds, store the spawn in flat bottles, sterilize equipment using an autoclave, secure the mixture inside the bottle with cotton balls, and cover the mixture with foil or paper. Procedure: 1. Wash the sorghum until the water is clear 2. Boil sorghum grains until it is half cooked (not so soft but already cooked)-30 minutes 22 3. Drain the sorghum grains and let it cool. 4. Put sorghum grains in the flat bottle about ¾ full. 5, Properly seal the bottle and sterilized it on a pressure cooker/ autoclave at 15 psi for 30 minutes. 6. After sterilization, collect the bottle and let it cool and ready for inoculation. 23  C.Inoculation of Spawn Materials: In inoculating spawn, the researcher uses a pure culture mushroom, an alcohol lamp to sterilize tools and create a sterile environment, alcohol as a disinfectant to clean hands and tools, a wire loop to transfer the mushroom culture to a sterile tool, and an inoculation chamber to reduce the possibility of contamination. Procedures: 1. Sanitized all materials need to avoid contamination. 24 2’ Using a mushroom picker/ wire loop cut the mycelia on the pure culture at about 1 sq. inch  3. Pick the cut mycelia and put it on the bottle of the mother spawn 4. Seal properly the inoculated mother grain spawn and put it on a dark room for incubation. 27  OYSTER MUSHROOM CULTIVATION A. Collecting of Materials/Substrate Preparation Materials: In order to prepare the substrate and gather materials, the researcher first gathers 83 kl of sawdust, 15 kl of rice bran, 1 kl of sugar, 1 kl of lime, 1 small pale of water, a hovel or pallet to mix all the materials, rice sacks to cover the materials, cellophane (6x12x.02) to hold the substrate, a rubber band to tie the cellophane, and finally a drum to sterilize the fruiting bags for 6–8 hours. Procedures: 1. Collecting of sawdust substrate from inside/outside school perimeter 3. Weight all the ingredients using weighing scale. 2. Mix the sawdust, rice bran, limestone, brown sugar and water using shovel and add 10 pale of water.  4. Ferment the substrate to 5-7 days. Mix the substrate every two days. 5. Bag the substrate into cellophane after 7 days of fermentation. Close it using rubber band. 6. Sterilize the fruiting bags inside the drum for 6-8 hours. 7. Cool the fruiting bags for 24 hours before inoculation. A. Spawning/Planting Procedures Materials: In the Spawning/Planting Procedures, the researchers used fruiting bags to keep all the ingredients inside, Pleurotus ostreatus spawn, which is the primary material used to grow mushrooms, 70% alcohol to keep the spawn from becoming contaminated, a candle to keep insects out of the spawn, cotton balls to balance the air coming from the outside, rubber bands to tie the fruiting bag, a match to light the candle, and finally a spoon to collect the spawn. Procedures: 1. Prepare all the materials needed 2. Open the fruiting bags 3. Inoculate the spawn of P. ostreatus  4. Put the cotton inside the opening of the cellophane and put a rubber band. 5. Inoculate the fruiting bags for 1 to 2 months for colonization C.Spraying Procedures Materials: The researcher employed a variety of mixtures in the spraying procedures, including clean water (Control), rice wash (Treatment 1), water and salt (Treatment 2), water and sugar (Treatment 3), water and Probiotic milk (Treatment 4), and a sprayer in which the various treatments were placed. Procedures: VARYING WATER MIXTURES AS FERTILIZER SUPPLEMENT Spraying of Oyster Mushroom Pleurotus ostreatus using different water mixtures as fertilizers supplement, 3 times a day for 1 month observation The fruiting bags are treated with five (5) different water mixtures as fertilizer supplement, the water (control), rice wash (treatment 1), water and salt (treatment 2), water and sugar (treatment 3) and water and probiotic milk (treatment 4). 1st DAY 2nd DAY 3rd DAY  C.Harvesting and Weighing Procedures Materials: In harvesting and weighing procedures, the researcher used scissors to cut the mushrooms, cellophane to collect the mushrooms after harvesting, a plastic tray to gather all the mushrooms, and lastly, a weighing scale to measure the kilograms of mushrooms. Procedures: 1. Harvest the fruits of mushrooms P. ostreatus by each treatments 2. Separate the fruit of mushrooms P. ostreatus by treatments 3. Weigh the mushroom P. ostreatus by each treatments 4. Get the exact weight of harvested mushroom P. ostreatus by each treatments Statistical Treatment The researchers used statistical tools appropriate to determine the significant difference of varying water treatments and fruiting bags. Statistical tools used were mean difference and One-Way ANOVA to determine significant difference between fruiting bags and shows the effectiveness of under different water mixtures.  RESULTS AND DISCUSSIONS Table 1. presents the Flower Production of Oyster mushroom Pluerotus osteatus on different fruiting bags treated with varying water mixtures as fertilizer supplement. Table 1. Flower production of Oyster Mushroom Pluerotus Ostreatus among Water Treatments Treated on Different Fruiting WEEK Grand Treatments 1 2 3 4 Control 1.7 1.1 14 2 12 1 14 2 8 4 1.71 Water Treatment 1 4.1 4.7 0.8 29 4 33 1 28 4 9 6 3.43 Rice wash Treatment 2 0.7 0.4 7 1 5 1 7 1 3 3 0.79 Water & Salt Treatment 3 2.7 2.5 4.4 19 1 18 7 31 3 7 1 2.68 Water & Sugar Treatment 4 4.7 3.8 6.1 1.8 33 4 27 6 43 4 13 6 4.15 Water & Probiotic milk Bags Among the experimental groups, Treatment 4, which involved the use of water and probiotic milk, resulted in the highest number of flowers. However, Treatment 1, which utilized rice water, had a mean value of 3.43 for flower production. Treatment 3, which involved water and sugar, had a mean value of 2.68, while the control group (Treatment 0) with water only had a mean value of 1.71. Treatment 2, which used water and salt, had the lowest number of fruits produced, with a mean value of 0.79.  Based on these results, it can be concluded that the combination of water and probiotic milk was the most effective in providing nutrients to influence the growth of Oyster mushrooms, particularly in increasing the number of flowers. Rice water also showed promising results in flower production, while water and sugar were less effective. The control group with water only had intermediate flower production, and Treatment 2 with water and salt had the lowest fruit production. The findings suggest that incorporating water and probiotic milk as supplements can greatly enhance the growth and reproductive performance of Oyster mushrooms. This implies that the addition of probiotic milk, which contains beneficial bacteria, and the use of rice water as nutrient sources can positively influence the cultivation process. The increased number of flowers indicates a higher potential for fruit production, leading to improved overall yield. Additionally, the negative impact of salt on mushroom growth and development highlights the importance of avoiding excessive salt content in the substrate. Overall, these findings contribute to optimizing mushroom cultivation techniques and promoting sustainable practices in the industry. In a study by Tiara in 2021, it was found that spraying a probiotic milk and water solution resulted in better growth of oyster mushrooms compared to using plain water. The average harvest frequency using the probiotic milk and water solution was 0.83 times, with an average wet weight of 22.6 grams per harvest, while the average harvest frequency using plain water was 0.66 times, with an average wet weight of 6.6 grams per harvest. These findings suggest that incorporating a probiotic milk and water solution through spraying can significantly improve the growth and yield of oyster mushrooms. The specific blend of ingredients and live bacteria in the probiotic milk appears to provide beneficial nutrients and create favorable conditions for mushroom growth (Tiara, 2021). Table 2. presents the ANOVA table on the Significant Difference in the Treatment Means on weight level of flowers among fruiting bags treated with different water mixtures on Oyster Pleurutos ostreatus mushroom. Table 2. ANOVA table on the Significant Difference in the Treatment Means on weight level of flowers among fruiting bags treated with different water mixtures on Oyster mushroom Pleurutos ostreatus Source of P- SS Df MS f F crit Variation value Between 718.76 3 239.59 1.13 0.66 3.49 Groups Within Groups 5099.24 24 212.47 Total 5818 27 The analysis of the data revealed that the computed f value of 1.13 was lower than the tabular f value of 3.49 at a significance level of 0.05, with 3 and 24 degrees of freedom. This indicates that there is no significant difference in the mean weights of flowers among the fruiting bags treated with different water mixtures as fertilizer supplements for the Oyster mushroom Pleurotus ostreatus. After four weeks of observation, the oyster mushroom growth performance across the different water mixtures resulted in a similar average number of flowers in the fruiting bags. The analysis showed that there was no significant difference in the weight of flowers among the Oyster mushroom Pleurotus ostreatus fruiting bags treated with different water mixtures as fertilizer supplements. This implies that the choice of water mixture does not significantly affect the growth performance in terms of flower weight. Growers may not need to focus extensively on selecting specific water mixtures fm or optimal flower production, but instead can prioritize other cultivation factors. However, further research is needed to explore the potential impact of water mixtures on other growth parameters.  Figure 4. Comparison of Treatment Means Figure 4 illustrates the growth performance of Oyster mushrooms in terms of weight (in grams) across different water treatments. Treatment 4, which involved the use of water and probiotic milk, resulted in the highest weight of 191.5 grams. Among the experimental groups, Treatment 3 (Water and sugar) had an average weight of 188.25 grams, Treatment 1 (Rice wash) had an average weight of 153.75 grams, the Control group (Water) had an average weight of 122.75 grams, and Treatment 2 (Water and salt) had the lowest weight with an average of 48 grams. These findings suggest that the water and probiotic milk mixture was the most effective in providing nutrients and serving as a fertilizer supplement to promote the growth of Oyster mushrooms, particularly in terms of weight gain. The implication of the findings presented in Figure 4 is that the choice of water mixture as a fertilizer supplement can have a significant impact on the growth performance and weight of Oyster mushrooms (Pleurotus ostreatus). Treatment 4, which involved the use of water and probiotic milk, resulted in the highest weight of 191.5 grams. This suggests that incorporating probiotic milk in the water mixture can provide essential nutrients and promote optimal growth conditions for the mushrooms. This finding has practical implications for mushroom growers, as they can enhance the weight and overall yield of Oyster mushrooms by utilizing a water and probiotic milk mixture as a fertilizer supplement. It highlights the importance of considering the specific composition of the water mixture to maximize the growth potential of Oyster mushrooms. Conclusions 1. Treatment 4, which involved the use of water and probiotic milk, resulted in the highest number of flowers, making it the most effective fertilizer supplement for growing Oyster Pleurotus ostreatus mushrooms. 2. Treatment 4 also yielded the heaviest weight of mushrooms, indicating that it was the most effective fertilizer supplement for promoting the overall growth and weight gain of Oyster Pleurotus ostreatus mushrooms. 3. The analysis showed that there was no significant difference in the weight of flowers among the fruiting bags treated with different water mixtures. This suggests that the choice of water mixture did not significantly affect the flower weight in Oyster Pleurotus ostreatus mushrooms. 4. Overall, among the different water mixtures used as fertilizer supplements, Treatment 4 (Water and Probiotic Milk) emerged as the most effective nutrient enhancer for the growth of Oyster Pleurotus ostreatus mushrooms, as it resulted in both higher flower numbers and greater weight. Recommendations 1. Prioritize Treatment 4 (Water and Probiotic Milk) as the recommended fertilizer supplement for Oyster Pleurotus ostreatus mushroom cultivation due to its superior results in flower production and weight. 2. Conduct further research to understand the specific nutrients and beneficial bacteria in probiotic milk that contribute to enhanced mushroom growth, optimizing its formulation and application. 3. Incorporate nutrient-rich water mixtures in cultivation practices to maximize Oyster Pleurotus ostreatus mushroom growth and yield. 4. Monitor and adjust watering practices to maintain proper moisture levels for optimal mushroom development. 5. Promote sustainable cultivation methods, such as using recycled water and incorporating organic waste, to reduce water consumption and minimize environmental impact in Oyster Pleurotus ostreatus mushroom cultivation.  BIBLIOGRAPHY Cheung, P.C.K. (2010). The nutritional and health benefits of mushroom. Nutrition Bulletin, 4(35), 292-299. Dobermann, A., & Fairhurst, T. H. (2002). Rice straw management. Better Crops International, 16(1), 7-11. Fan, K., Chen, L., Cai, J., Liu, S., Li, Y., & Shen, Y. (2011). Preliminary study on oyster mushroom growth situation in 12 kinds of straw substrates. Journal of Anhui Agricultural University, 38(5), 806-811. Győrfi, J., & Hajdu, C. S. (2007). Casing-material experiments with Pleurotus eryngii. 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Use of wheat straw residue from a paper mill for Pleurotus ostreatus cultivation. Acta Edulis Fungi, 17(4), 30- 31. 45  ACKNOWLEDGEMENT This research project could not have been completed successfully without the invaluable assistance and support of Mrs. Christy S. Bagayas, the Secondary School Principal II of New Panay National High School, for her constant support. Additionally, the researchers want to express their sincere gratitude to their research adviser, Dr. Melvin A. Garcia, MT-I. His constant direction, and inspiring nature were crucial to this project's successful conclusion. The researchers would like to express their profound gratitude to their supportive parents, namely Mr. & Mrs. Neri, Mr. & Mrs. Panada, and Mr. & Mrs. Reyes. Their financial support, understanding, and unwavering prayers were essential sources of strength and inspiration throughout this journey. For their innovative effort in this study, researcher would like to express their profound gratitude to the original authors, Mormolindo et al. Their findings have greatly advanced our knowledge and given us a strong starting point for our own research. 46  Finally, the researchers thank and acknowledge the Mighty Creator, their Heavenly Father, without whom this study would not have been possible. They sincerely thank Him and give Him honor for giving them their strength and wisdom. The Researchers 47