Enhancing Sustainable Agriculture through Wastewater Utilization and Low-Cost Organic Fertilizer Production

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The study explores the cultivation of nitrogen-rich sources such as Spirulina and Azolla using wastewater as a growth medium, aiming to reduce production costs and address water scarcity problems. Additionally, a low-cost organic fertilizer (LOF) was developed using underutilized resources like Palmyrah leaves, Coconut leaves, and Banana pseudostem, with Spirulina, and Azolla aiming to counteract the detrimental effects of inorganic fertilizers and contribute to the circular economy by converting waste into valuable agricultural inputs. Methods: Experimental trials assessed the efficacy of wastewater as a growth medium for Spirulina and Azolla, alongside evaluating the impact of organic nutrient sources on seed germination and seedling vigor. Pot experiments were conducted to assess biomass yield and crop growth parameters across various fertilizer treatments, including conventional organic, LOF, and inorganic methods. Results: Results reveal that treatments containing wastewater significantly enhanced the biomass yield of Spirulina and Azolla compared to standard methods, offering a cost-effective solution. The LOF enhanced the seed germination (5-15%), and seedling vigor (12-53%) of selected seeds compared to the control. Pot experiment revealed that the combination treatments of LOF gave improvements in crop growth parameters, biomass yield, and nutrient content of Sugargraze (Hybrid sweet sorghum) compared to conventional organic or inorganic treatments. Notably, substituting 50% of organic or inorganic fertilizer with LOF gave comparable or higher performance, highlighting the potential of LOF as an environmentally sustainable alternative. Conclusion: Overall, this study underscores the feasibility of integrating underutilized organic nutrient sources and wastewater utilization in sustainable agriculture. Azolla Crop productivity Integrated plant nutrition system Organic farming Wastewater utilization Spirulina Figures Figure 1 Figure 2 Introduction Fertilizers have been an indispensable input in agriculture in enhancing crop yields and market value. However, the excessive use of inorganic fertilizers by farmers has led to detrimental environmental consequences [ 1 , 2 ]. Inorganic fertilizers are hazardous to living beings due to the bio-magnification of toxic elements from inorganic fertilizers which accumulate in the plant and lastly reach our digestive system [ 3 , 4 ] and also affect the soil biota [ 5 ]. Available nutrients from inorganic fertilizers especially nitrate easily leach away into groundwater without fully benefiting the plant and cause groundwater pollution [ 6 ]. Increasing the rate of inorganic fertilizer application tended to reduce the carbon sequestration level and emit CO 2 into the environment which leads to the greenhouse gas effect [ 7 ]. To mitigate these issues, there has been a growing trend in transitioning towards sustainable agricultural practices such as organic farming and integrated plant nutrition systems (IPNS) [ 8 ]. Organic fertilizers are a promising alternative to conventional inorganic fertilizers especially organic waste as fertilizer is proposed as a sustainable alternative to mineral fertilizer [ 9 ] offering environmental benefits as well as economic viability. Organic manures have multiple benefits including the potential for preserving soil quality, enhancing carbon sequestration, minimizing water usage, preserving biodiversity, and ensuring the production of healthy food [ 10 ]. Despite these advantages, there exists a gap in knowledge among farmers regarding the potential utilization of underutilized organic residues as nutrient sources. Organic materials such as Palmyrah ( Borassus flabellifer L.) leaf [ 12 ], Coconut ( Cocos nucifera L.) leaf [ 11 ], and Banana ( Musa acuminata ) pseudostem [ 13 ] have been reported to have high nutrient content and are abundantly available in farms and home gardens, remain underutilized due to their low nitrogen content. Azolla, a nitrogen-fixing aquatic fern [ 14 ], and Spirulina, a cyanobacterium, offer viable alternatives with high nitrogen content and rapid growth rates [ 15 , 16 ]. Both are rich in phytohormones and play an important role in the growth and development of plants as regulators [ 17 ]. However, their cultivation requires substantial water resources, posing challenges in regions facing water scarcity. Additionally, large-scale production of Spirulina necessitates significant inputs of chemicals for the standard medium preparation, contributing to production costs. Approximately 80% of the total costs for microalgae cultivation are associated with excessive consumption of nutrients and water [ 18 ]. Addressing these difficulties, this research focused on exploring the potential of utilizing wastewater as a medium for cultivating nitrogen-rich sources such as Spirulina and Azolla and integrating the underutilized nutrient sources namely palymyrah leaf, coconut leaf, and banana pseudostem to develop a low-cost organic fertilizer (LOF) satisfying the nutrient requirements of a commercial organic fertilizer. The LOF was evaluated for its impact on seed germination and seedling vigor of selected seeds and crop performance of Sugargraze ( Sorghum bicolor (L.) Moench ) exploring the substitution of 50% of organic or inorganic fertilizer with LOF. Methodology Wastewater Preparation and Semi-mass Culturing of Spirulina To initiate the study, samples of reverse osmosis (RO) wastewater and parboiled effluent were collected from the University RO plant at Ariviyalnagar and Killinochchi rice mill, respectively. These samples were subjected to initial processing via sand filtration to reduce the turbidity concerns. In a complete randomized design, the experiment was structured with four treatments, denoted as T1 to T4. T1 represented the control group with 100% Zarrouk’s medium, while T2 involved a dilution of Zarrouk’s medium to 50%. T3 and T4 comprised combinations of 50% Zarrouk’s medium with 50% parboiled effluent and 50% RO wastewater, respectively. Each treatment was replicated thrice to ensure the reliability of the results. A two-liter formulated low-cost medium was prepared and inoculated with a 100 ml culture of Spirulina ( Spirulina subsalsa ) at a 0.5 optical density, sourced from a two-week-old fresh culture. The cultivation setup was placed under 65% shade within a net house environment. Regular readings were recorded until a consistent optical density (OD) was achieved, signaling the maturity of the Spirulina biomass. Following this, the biomass was harvested, filtered using Whatman No. 42 filter paper, and subjected to oven drying at 60°C for a duration of 48 hours. Azolla Cultivation A complete randomized design with nine treatments (T1 - Kitchen wastewater + triple super phosphate (TSP), T2- Tap water + TSP, T3- RO wastewater + TSP, T4 - Kitchen wastewater + Cow dung, T5 - Tap water + Cow dung, T6 - RO wastewater + Cow dung, T7 -Kitchen wastewater, T8 - Tap water, T9 - RO wastewater) and triplicates was prepared for Azolla cultivation to ensure statistical significance and reproducibility. Tanks (30×15 cm) were filled with a 10 cm depth of selected water. Eleven g of 2 days decomposed cow dung and 0.5 g triple super phosphate (TSP) was added with selected water based on decided treatments [ 19 ]. The setup was kept for 2 days to settle down the particles. 0.1 g of fresh Azolla biomass was crushed and added to each tank separately. The setup was kept under 65% shade net house condition. After 14 days biomass was harvested and oven dried at 60 0 C for 48 hours to reach the constant weight and dry weight was measured. Analysis of Organic Sources and Plant Tissue Organic sources were prepared through a sequence of chopping, washing, drying, grinding, and sieving through a 2 mm sieve. The powdered samples were stored in airtight conditions for nutrient analysis. The determination of total nitrogen, phosphorus, potassium, and total organic carbon content was carried out employing established laboratory standard methodologies such as total nitrogen (Kjeldhal method [ 20 ]), phosphorus (Vanadomolybdate method [ 21 ]) potassium [ 22 ], total organic carbon [ 23 ] and Calcium and magnesium were measured using an atomic absorption spectrophotometer. Formulation of LOF LOF was formulated by mixing 25% Banana pseudostem, 25% Palmyrah leaf, 25% Coconut leaf, 1% Spirulina, and 24% Azolla powders based on availability, nutrient analysis and C/N ration to achieve the level of N (1.47%), P (0.34%), K (1.12%) and C/N ratio (24.66) as equal or higher than that of the organic fertilizer, cattle manure N (1.42%), P (0.31%), K (1.21%) and C/N ratio (23.42). Germination and Vigor Test Seeds were soaked for 3 hours with 2% of seed weight in an organic source. Curry Chilli and Green Chilli seeds were placed in filter paper, while Water spinach and Sugargraze seeds were placed in sand beds [ 24 ]. Germinated seeds were counted at 2-day intervals. Seedling height was measured, and germination percentage and seedling vigor were calculated based on standard procedures. The percentage of germination was calculated by dividing the number of seeds germinated by the total number of seeds sown [ 25 ]. Percentage of germination = (Number of seeds germinated / Number of seeds sown) × 100% Seedling height (cm): Three seedlings were randomly uprooted from each replicate in each treatment and height was measured in centimeters. The Vigour index was estimated using the following formula (Sharma et al., 2014). Vigour index = (Seedling height (cm) × Germination percentage) Pot Experiment The pot experiment focused on Sugargraze as the test crop and was organized in a complete randomized design (CRD) comprising seven distinct treatments denoted as T1 to T7. These treatments included T1 - Control, T2–100% IN, T3–50% IN, T4–100% CM, T5–50% CM, T6–50% IN + 50% LOF, T7–50% CM + 50% LOF (IN: Inorganic fertilizer based on department recommendation, CM: cattle manure, LOF: low-cost organic fertilizer). Black polythene bags were used as pots with sieved soil. Organic treatments received cattle manure and low-cost organic fertilizer, inorganic treatments received urea (150 Kg/ha), Muriate of potash - MOP (50 Kg/ha), and Triple super phosphate - TSP (100 Kg/ha) fertilizers and they were applied at the Department of Agriculture (DOA) Sri Lanka recommended rates. Data Collection and Statistical Analysis Data on growth and yield parameters were collected, and statistical analysis was performed using the Statistical Analytical System (University version) with Duncan's mean separation at P = 0.05. Result and Discussion Biomass Yield of Spirulina The efficiency of semi-mass culturing was measured in terms of biomass yield, expressed in dry weight per unit volume. In this study, treatment with 50% Zarrouk’s medium + 50% parboiled effluent (T3) exhibited a significantly higher dry biomass yield compared to treatment with 100% Zarrouk’s medium (Table 1 ). It was observed that treatments with 100% Zarrouk’s medium (T2) and 50% Zarrouk’s medium + 50% RO wastewater (T4) did not exhibit significant differences in dry biomass. Notably, 50% Zarrouk’s medium + 50% parboiled effluent (T3) gave the highest dry biomass (3.023 g/l) of Spirulina on the 30th day of cultivation because the parboiled effluent, rich in ammonia-nitrogen, may have contributed to the increase in biomass yield in T3. Parboiled rice effluent is known to contain high levels of organic matter, nitrogen, and phosphorus [ 26 ] while RO wastewater comprises both organic and inorganic components, including single-charged ions such as Na + and Cl − [ 27 ]. Treatment combinations involving parboiled effluent and RO wastewater provided the necessary nutrient levels for Spirulina growth, resulting in higher biomass yields. The utilization of 50% wastewater, such as parboiled effluent or RO wastewater, as a substitute for Zarrouk’s medium, was found to halve the production costs needed for medium because in the conventional process of Spirulina biomass production, cultivation utilizing chemical-based culture mediums accounts for 35% of the total production cost. Furthermore, the environmental impact during the cultivation stage is notably the most significant among all production stages, primarily due to the extensive use of chemicals and nutrients [ 28 ]. Additionally, treatments involving wastewater sources (T3 and T4) yielded higher biomass compared to T2 (50% Zarrouk’s medium alone), suggesting that wastewater serves as a nutrient supplement for Spirulina cultivation. Therefore, employing wastewater instead of clean water for Spirulina cultivation presents a viable solution to address water scarcity issues. According to Richmond (2004), the biomass production of Spirulina relies heavily on factors such as temperature and nutrient availability in the growing medium. The synthetic standard growing medium for spirulina cultivation is Zarrouk's medium, which is rich in essential elements such as sodium, potassium, calcium, magnesium, and traces of iron and manganese [ 29 ]. However, the cost of Zarrouk's medium is relatively high, and it typically requires laboratory-grade clean water for preparation, contributing to operational expenses [ 30 ]. In a previous study, Mukherjee et al., 2016 reported that microalgae cyanobacteria consortium was grown successfully in parboiled effluent and the biomass was used as fertilizer. In another study [ 41 ] rice mill wastewater in combination with sodium chloride was used to grow Spirulina. Usharani et al., 2012 used parboiled effluent in combination with sodium bicarbonate. Table 1 Treatments Scheduled for Spirulina and Azolla Semi - Mass Culturing, Recorded Data Treatments Spirulina Azolla Treatment Combination Dry Biomass Yield (g/l) Treatment combination Dry Biomass Yield (g/l) T1 100% Zm 2.81 b Kw + TSP 2.14 bc T2 50% Zm 1.45 d Tw + TSP 1.21 d T3 50% Zm + 50% Parboiled effluent 3.20 a RO + TSP 1.97 c T4 50% Zm + 50% RO 2.8 b Kw + Cd 2.22 b T5 - - Tw + Cd 2.09 bc T6 - - RO + Cd 3.14 a T7 - - Kw 0.60 e T8 - - Tw 0.57 e T9 - - RO 0.64 e The same letters within columns are not statistically different by the DUNCAN at p = 0.05 (Kw - Kitchen wastewater, Tw - Tap water, RO - Reverse osmosis wastewater, Cd - Cow dung, TSP - Triple super phosphate Zm - Zarrouk’s medium) Azolla Fresh and Dry Biomass Yield Table 1 presents the dry biomass yield of Azolla. Among all treatments, RO wastewater + cow dung (T6) obtained the highest dry biomass yield and it was significantly higher than other treatments. Next to T6 (RO wastewater + cow dung) T4 (Kitchen wastewater + Cow dung) gave high dry biomass yield. The treatments that includes wastewater components shows higher or equal dry biomass yield compared to treatments that includes tap water (fresh water). Treatments combined with cow dung gave a high dry biomass yield compared with other treatments. The results of this study reveal the potential of utilizing wastewater as a viable growth medium for Azolla cultivation, a practice not commonly explored in traditional cultivation methods where Azolla is typically grown in well water, pond water, and freshwater except few studies where Azolla was used to remediate wastewater rich in ammonium and phosphorus [ 31 ] The significant increase in dry biomass yield observed in treatments incorporating wastewater components underscores the efficacy of waste utilization in enhancing Azolla growth. Nutrient Analysis of Organic Sources Five different organic sources, cattle manure and LOF were analyzed. Table 2 presents the primary nutrients (C, N, P, and K) and secondary nutrients (Mg and Ca) available in organic sources. Among the organic sources, Spirulina recorded the highest N content (115.03ppm), followed by Azolla, Coconut Leaf, Palmyrah Leaf, Banana Pseudostem, and Cattle Manure. Banana pseudostem gave the highest P (6.17ppm) content, while Spirulina has the lowest. Banana pseudostem also contains the highest K content (19.56ppm), whereas Coconut Leaf has the lowest. Palmyrah Leaf has the highest C (832.50 ppm) content, followed by Coconut Leaf, Banana Pseudostem, and Spirulina. Among secondary nutrients, Azolla recorded the highest amount of Ca (10861ppm) and Mg (1947.2ppm). The LOF is formulated to match the nutrient content of the prevailing organic source, in this case, cattle manure, in a proper ratio. By formulating LOF to have nutrient content equivalent to cattle manure, it ensures a consistent supply of nutrients for plant growth, utilizing locally available resources effectively. This approach promotes sustainable agriculture by utilizing organic waste materials as nutrient sources. Table 2 Nutrient Content of Organic Sources Organic Sources Primary Nutrients Secondary Nutrients N (ppm) P (ppm) K (ppm) C (ppm) Mg (ppm) Ca (ppm) Coconut leaf 1.87 e 3.56 c 7.19 c 430 b 901.6 d 4099.2 b Spirulina 115.03 a 1.26 d 4.97 e 415 b 1919.55 ab 2134.4 c Azolla 32.43 b 1.06 e 14.09 b 317.5 c 1947.2 a 10861 a Palmyrah leaf 11.43 c 4.07 b 6.15 d 832.50 a 1348.38 c 3423.47 b Banana pseudostem 7.00 d 6.17 a 19.56 a 330 c 1665.6 b 184 d Cattle Manure 14.2 3.1 12.1 332.5 LOF 14.7 3.4 11.2 362.5 The same letters within columns are not statistically different by the DUNCAN at p = 0.05 Germination Percentage and Vigour Index Table 3 shows the effect of different treatments on germination percentage and vigor index. The germination test conducted for Curry Chilli, Water spinach, Green Chilli, and Sugargraze indicated that the utilization of Spirulina, Azolla, and LOF as soaking agents resulted in enhanced germination rates compared to the control group. Moreover, seedling vigor parameters exhibited notable improvements across all treatments when compared to the control (T5). Compared to other treatments T3 and T4 gave high germination percentages and vigor index; the production of growth substances and vitamins by the algae may be partly responsible for this [ 32 ]. Spirulina, Azolla, and Banana contain phytohormones like Gibberellins, Auxin, and Cytokinin itself [ 33 ]. Gibberellin can break the seed dormancy and increase the germination percentage [ 34 ], which could have contributed to higher germination percentage and germination vigor recorded in all treatments than control. Consequently, the application of Spirulina, Azolla, and LOF demonstrated positive effects on both seed germination and vigor index, suggesting their viability as commercially and ecologically sustainable biostimulants. Table 3 Effect of different treatments on germination percentage and germination vigor of selected seed Treatments Water spinach Curry Chilli Green Chilli Sugargraze G.P V.I G.P V.I G.P V.I G.P V.I T1 94% 1495.21 b 96% 909.34 a 98% 662.07 ab 97% 2201.11 a T2 91% 1441.61 b 98% 775.72 b 96% 611.91 b 97% 2008.89 a T3 96% 1614.09 a 96% 923.34 a 100% 691.11 a 97% 2258.89 a T4 93% 1611.63 a 99% 882.12 a 99% 669.34 ab 100% 2288.89 a T5 87% 1137.00 c 93% 692.30 c 94% 509.88 c 87% 86.6667 b T1 - Low-cost organic fertilizer, T2 - Dry Azolla powder, T3 - Dry Spirulina Powder, T4 - Live Spirulina Culture, T5 - Control (Distilled water) G.P - Germination Percentage, V.I – Vigor Index Pot Experiment - Sugargraze Plant Height The plant height at 2-week intervals is illustrated in Table 4 . Almost during all weeks significantly higher plant height was observed in T2 (100% inorganic) and T6 (50% inorganic + 50% LOF) than other treatments. However, there were no significant height differences between T2 (100% inorganic) and T6 (50% inorganic + 50% LOF). During 10th and 12th weeks after seeding the height difference among treatments was similar. It is significant to note that plant height in T7 (50% organic + 50% LOF) was significantly higher than that of T4 (100% organic) after 4th week. Table 4 Plant height - Sugargraze Treatments 2nd week 4th week 6th week 8th week 10th week 12th week T1 28.00 c 36.50 c 48.50 e 75.00 c 102.00 f 121.00 f T2 41.50 a 68.00 a 120.00 ab 172.50 a 191.00 a 210.00 a T3 35.00 ab 58.50 b 101.50 c 142.50 b 161.00 d 183.00 d T4 31.00 bc 57.00 b 104.00 c 147.50 b 181.00 c 193.50 c T5 29.75 bc 40.50 c 62.00 d 95.00 c 141.00 e 150.50 e T6 41.50 a 67.5 a 124.50 a 171.50 a 192.50 a 210.00 a T7 34.50 bc 57.50 b 111.00 bc 157.50 ab 186.00 b 195.50 b T1 - Control, T2–100% inorganic, T3–50% inorganic, T4–100% organic, T5–50% organic, T6–50% inorganic + 50% LOF, T7–50% organic + 50% LOF Fresh and Dry Biomass Weight Figure 1 shows the fresh and dry biomass weight of plants with different treatments. The high fresh weight per plant was observed in T4 (100% organic) and T7 (50% organic + 50% LOF) while the lowest was in T1 (Control). Fresh weight of T4 (100% organic), T6 (50% inorganic + 50% LOF), and T7 (50% CM + 50% LOF) were not statistically significant among each other. The higher dry weight per plant was observed in T4 (100% organic) and T7 (50% organic + 50% LOF) while the lowest was in T1 (Control). There were no significant dry biomaáss differences among T2 (100% inorganic), T4 (100% organic), T6 (50% inorganic + 50% LOF), and T7 (50% organic + 50% LOF). Nutrient Content (N, P, K) - Sugargraze Figure 2 shows the nutrient content in plants with different treatments. The high N uptake was recorded with T6 (50% inorganic + 50% LOF) and T7 (50% organic + 50% LOF) and it was significantly higher than other treatments. The lowest N uptake was noted in T1 (Control).The high P uptake was observed in T4 (100% organic) & T7 (50% organic + 50% LOF). P uptake in T4 (100% organic) & T7 (50% organic + 50% LOF) were significantly higher than other treatments however there were no significant differences among them. The low P uptake was recorded with T1 (Control) and T3 (50% inorganic). K uptake in the plant was high in T4 (100% organic), T6 (50% inorganic + 50% LOF), and T7 (50% organic + 50% LOF). In accordance with N, P, and K the sole organic treatments namely T4 (100% organic) & T7 (50% organic + 50% LOF), and IPNS T6 (50% inorganic + 50% LOF) had a higher % of nutrient content than that of T2 (100% inorganic) because of high nutrient leaching problem in 100% inorganic system even though it has high nutrient availability [ 35 , 36 ]. In the pot experiment, treatments T2 (100% inorganic), T6 (50% inorganic + 50% LOF), and T7 (50% organic + 50% LOF) exhibited significantly higher values in crop growth parameters, dry biomass yield, and nutrient content in comparison to other treatments. Notably, there were no significant differences observed among these three treatments. Significantly higher growth and yield of Wheat and nutrient uptake in combined organic-inorganic treatments than sole inorganic treatment [ 37 , 38 ], better yield performance and quality of Mustard in integrated nutrient treatments [ 39 ] better performance of integrated nutrient management in different crops also reported by Fan et al., 2008 and Selim 2020. From an environmental and health perspective, the combination of 50% inorganic fertilizer with LOF (T6) or the sole organic combination with LOF (T7) emerged as viable alternatives to the 100% inorganic treatment (T2). The integrated nutrient management tool efficiently supplies plant nutrients, lowers costs, improves soil conditions, promotes a healthy environment, maintains nutrient balance, and manages agricultural waste safely [ 40 ]. The findings of the present study underscores that substituting 50% of organic or inorganic fertilizer with LOF can yield comparable or higher performance in sugargraze cultivation. CONCLUSION The research findings reveal the potential of Spirulina, Azolla, and LOF as bio-stimulants in enhancing seed germination and vigor across multiple crops. Spirulina semi mass culturing experiments revealed that incorporating wastewater sources significantly enhanced biomass yield, offering a cost-effective solution and addressing water scarcity issues. Azolla cultivation experiments emphasized the potential of utilizing kitchen and RO wastewater in combination with cow dung, to achieve comparable or higher biomass yields compared to tap water in combination with cow dung. Importantly, underutilized organic sources and nitrogen sources from wastewater were identified as valuable alternatives to conventional fertilizers, Spirulina serves as a promising alternative source for urea, Azolla addresses secondary nutrient deficiencies (Ca, Mg), Coconut leaf and Palmyrah leaf act as valuable sources of carbon, and Banana pseudostem offers an alternative source for triple superphosphate and muriate of potash. In the pot experiment of sugargraze, combinations involving LOF gave notable improvements in crop growth parameters, biomass yield, and nutrient content compared to conventional inorganic treatments. Particularly, substituting 50% of organic or inorganic fertilizer with LOF proved beneficial from both environmental and health standpoints. These organic sources demonstrate significant potential to replace inorganic fertilizers in agricultural practices. These findings collectively advocate for the adoption of environmentally sustainable practices in agriculture, highlighting the feasibility of integrating organic nutrient sources to enhance crop productivity while minimizing environmental impact and resource consumption. Declarations Funding We affirm that we did not receive any funds, grants, or additional support during the preparation of this manuscript. Competing Interests The authors have no relevant financial or non-financial conflicts to disclose. Author Contributions Both authors contributed to the conception and design of the study. Material preparation, data collection, and analysis were conducted by Kasthuri Kajeevan. The initial draft of the manuscript was authored by Kasthuri Kajeevan and subsequently revised by Nalina Gnanaverajah. Both authors read and approved the final manuscript. 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(2019) Sharma, A.D., Rathore, S.V.S., Srinivasan, K., Tyagi, R.K.: Comparison of various seed priming methods for seed germination, seedling vigor, and fruit yield in okra (Abelmoschus esculentus L. Moench), vol. 165, pp. 75–81. Scientia horticulturae (2014) Tarekegn, N.M., Mengistie, E.: Physicochemical Characterization of Parboiled Rice Effluent: A Case Study in a Medium Scale Parboiled Rice Mill. J. Environ. Anal. Toxicol. 7 (5), 1–5 (2017) Lee, Y., Elimelech, M.: Chemical and Physical Aspects of Reverse Osmosis Desalination Pretreatment. Water Res. 40 (17), 3137–3160 (2006) Lim, H.R., Khoo, K.S., Chew, K.W., Chang, C.K., Munawaroh, H.S.H., Kumar, P.S., Huy, N.D., Show, P.L.: Perspective of Spirulina culture with wastewater into a sustainable circular bioeconomy. Environmental Pollution , 284 , p.117492. (2021) Raoof, B., Kaushik, B.D., Prasanna, R.: Formulation of a low-cost medium for mass production of Spirulina. 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Plant Sci. 52 (3), 295–303 (1972) Di, H.J., Cameron, K.C.: Nitrate leaching in temperate agroecosystems: sources, factors and mitigating strategies. Nutr. Cycl. Agrosyst. 64 , 237–256 (2002) Lehmann, J., Pereira da Silva, J., Steiner, C., Nehls, T., Zech, W., Glaser, B.: Nutrient availability and leaching in an archaeological Anthrosol and a Ferralsol of the Central Amazon basin: fertilizer, manure and charcoal amendments. Plant. soil. 249 , 343–357 (2003) Sarwar, M., Jilani, G., Rafique, E., Akhtar, M.E., Chaudhry, A.N.: Impact of integrated nutrient management on yield and nutrient uptake by maize under rain-fed conditions. Pakistan J. Nutr. 11 (1), 27–33 (2012) Selim, M.: Potential role of cropping system and integrated nutrient management on nutrients uptake and utilization by maize grown in calcareous soil. Egypt. J. Agron. 40 (3), 297–312 (2018) Tripathi, M.K., Chaturvedi, S., Shukla, D.K., Mahapatra, B.S.: Yield performance and quality in Indian mustard (Brassica juncea) as affected by integrated nutrient management. Ind. J. Agron. 55 (2), 138–142 (2010) Selim, M.: Introduction to the integrated nutrient management strategies and their contribution to yield and soil properties. International Journal of Agronomy, 2020. (2020) Amala, K., Ramanathan, N.: Chlorophyll production from Spirulina platensis (single cell protein, SCP); cultivation with sodium chloride in rice mill waste water. (2013) Fan, M., Cui, Z., Chen, X., Jiang, R., Zhang, F.: Integrated nutrient management for improving crop yields and nutrient utilization efficiencies in China. J. Soil Water Conserv. 63 (4), 126A–128A (2008). 10.2489/jswc.63.4.126a Richmond, A. (ed.): Handbook of Microalgal Culture: Biotechnology and Applied Phycology. Blackwell Publishing (2004) Usharani, G., Saranraj, P., Kanchana, D.: Spirulina cultivation: a review. Int. J. Pharm. Biol. Arch. 3 (6), 1327–1341 (2012) Supplementary Files Graphicalabstract.jpg HighlightsWBV.docx NoveltystatementWBV.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4411495","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":308358799,"identity":"965ac1ef-ba6d-45f0-8dd2-474ff38f16d9","order_by":0,"name":"Kasthuri Kajeevan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIie2SsWrDMBCGLwjsRXQ+40BeQaGgNNQkr+JgaJZAt5BunpRFoQ9TKB1jtDp7x5auHVyyGLr0ZGcKXLoWqm84wUmf9AsJIBD4gwysLyVAFFPJAYQfiRt1Uam8Ive94kcCWQXkSQHMT43fFGHdx/Hrxd1fJZ/67d1ALNNy0LSA12ywndFY1W5q0tVELQwFG+5FYgE1qzyChso4FaUrjXkNYk4JUwqW8Up8bDolqXtFYi6+Lyo7q7BTUJKy6ZTIn8IHs/UaD2apInm3JgVJWZgp7cNef2yXT82DuVWjrXtOWpUVEgv32m6ycckpZxNY9J+Bf0gYnTdm/NpAIBD4r/wAM71KfKC86aIAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0001-9853-9810","institution":"University of Jaffna Faculty of Agriculture","correspondingAuthor":true,"prefix":"","firstName":"Kasthuri","middleName":"","lastName":"Kajeevan","suffix":""},{"id":308358800,"identity":"6fdef27f-2d7c-4096-96a2-c258d55665cf","order_by":1,"name":"Nalina Gnanavelrajah","email":"","orcid":"","institution":"University of Jaffna","correspondingAuthor":false,"prefix":"","firstName":"Nalina","middleName":"","lastName":"Gnanavelrajah","suffix":""}],"badges":[],"createdAt":"2024-05-13 07:50:48","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4411495/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4411495/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58183987,"identity":"329a2ec8-8825-4623-89d4-4043e73ee140","added_by":"auto","created_at":"2024-06-12 06:56:16","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":29082,"visible":true,"origin":"","legend":"\u003cp\u003eFresh and Dry biomass yield - Sugargraze\u003c/p\u003e\n\u003cp\u003eT1 - Control, T2 - 100% inorganic, T3 - 50% inorganic, T4 - 100% organic, T5 - 50% organic, T6 - 50% inorganic + 50% LOF, T7 - 50% organic + 50% LOF\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/dbc6f9fa54f8cc6a52e314af.png"},{"id":58183984,"identity":"43fafed7-a357-478a-a5a5-cc8e3fe17c09","added_by":"auto","created_at":"2024-06-12 06:56:16","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":29789,"visible":true,"origin":"","legend":"\u003cp\u003eNutrient content - Sugargraze\u003c/p\u003e\n\u003cp\u003eT1 - Control, T2 - 100% inorganic, T3 - 50% inorganic, T4 - 100% organic, T5 - 50% organic, T6 - 50% inorganic + 50% LOF, T7 - 50% organic + 50% LOF\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/3d09a4bb1199abb2ee497ac4.png"},{"id":60191646,"identity":"46132f7b-d015-4a50-a21a-428564dd1aa8","added_by":"auto","created_at":"2024-07-12 20:52:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":686177,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/aa00b4ef-4be5-4f66-837e-bff47164f493.pdf"},{"id":58183985,"identity":"5b6c25e8-59f3-47fe-b427-db32b77d05b7","added_by":"auto","created_at":"2024-06-12 06:56:16","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":184768,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicalabstract.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/7000a92d30357fc327fee156.jpg"},{"id":58184429,"identity":"1721103d-301d-4113-bb68-314c2d830755","added_by":"auto","created_at":"2024-06-12 07:04:16","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":19758,"visible":true,"origin":"","legend":"","description":"","filename":"HighlightsWBV.docx","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/9645ed6d09f2bda5f90b3dfc.docx"},{"id":58183982,"identity":"0287b2b6-c7ee-4c45-907e-e18dd235c919","added_by":"auto","created_at":"2024-06-12 06:56:16","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":17676,"visible":true,"origin":"","legend":"","description":"","filename":"NoveltystatementWBV.docx","url":"https://assets-eu.researchsquare.com/files/rs-4411495/v1/9a4c18a1484cc738fe79e4f4.docx"}],"financialInterests":"","formattedTitle":"Enhancing Sustainable Agriculture through Wastewater Utilization and Low-Cost Organic Fertilizer Production","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFertilizers have been an indispensable input in agriculture in enhancing crop yields and market value. However, the excessive use of inorganic fertilizers by farmers has led to detrimental environmental consequences [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Inorganic fertilizers are hazardous to living beings due to the bio-magnification of toxic elements from inorganic fertilizers which accumulate in the plant and lastly reach our digestive system [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e] and also affect the soil biota [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Available nutrients from inorganic fertilizers especially nitrate easily leach away into groundwater without fully benefiting the plant and cause groundwater pollution [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Increasing the rate of inorganic fertilizer application tended to reduce the carbon sequestration level and emit CO\u003csub\u003e2\u003c/sub\u003e into the environment which leads to the greenhouse gas effect [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. To mitigate these issues, there has been a growing trend in transitioning towards sustainable agricultural practices such as organic farming and integrated plant nutrition systems (IPNS) [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Organic fertilizers are a promising alternative to conventional inorganic fertilizers especially organic waste as fertilizer is proposed as a sustainable alternative to mineral fertilizer [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] offering environmental benefits as well as economic viability. Organic manures have multiple benefits including the potential for preserving soil quality, enhancing carbon sequestration, minimizing water usage, preserving biodiversity, and ensuring the production of healthy food [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Despite these advantages, there exists a gap in knowledge among farmers regarding the potential utilization of underutilized organic residues as nutrient sources. Organic materials such as Palmyrah (\u003cem\u003eBorassus flabellifer\u003c/em\u003e L.) leaf [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], Coconut (\u003cem\u003eCocos nucifera\u003c/em\u003e L.) leaf [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], and Banana (\u003cem\u003eMusa acuminata\u003c/em\u003e) pseudostem [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] have been reported to have high nutrient content and are abundantly available in farms and home gardens, remain underutilized due to their low nitrogen content.\u003c/p\u003e \u003cp\u003eAzolla, a nitrogen-fixing aquatic fern [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], and Spirulina, a cyanobacterium, offer viable alternatives with high nitrogen content and rapid growth rates [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Both are rich in phytohormones and play an important role in the growth and development of plants as regulators [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. However, their cultivation requires substantial water resources, posing challenges in regions facing water scarcity. Additionally, large-scale production of Spirulina necessitates significant inputs of chemicals for the standard medium preparation, contributing to production costs. Approximately 80% of the total costs for microalgae cultivation are associated with excessive consumption of nutrients and water [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Addressing these difficulties, this research focused on exploring the potential of utilizing wastewater as a medium for cultivating nitrogen-rich sources such as Spirulina and Azolla and integrating the underutilized nutrient sources namely palymyrah leaf, coconut leaf, and banana pseudostem to develop a low-cost organic fertilizer (LOF) satisfying the nutrient requirements of a commercial organic fertilizer. The LOF was evaluated for its impact on seed germination and seedling vigor of selected seeds and crop performance of Sugargraze (\u003cem\u003eSorghum bicolor\u003c/em\u003e (L.) \u003cem\u003eMoench\u003c/em\u003e) exploring the substitution of 50% of organic or inorganic fertilizer with LOF.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eWastewater Preparation and Semi-mass Culturing of Spirulina\u003c/h2\u003e \u003cp\u003eTo initiate the study, samples of reverse osmosis (RO) wastewater and parboiled effluent were collected from the University RO plant at Ariviyalnagar and Killinochchi rice mill, respectively. These samples were subjected to initial processing via sand filtration to reduce the turbidity concerns. In a complete randomized design, the experiment was structured with four treatments, denoted as T1 to T4. T1 represented the control group with 100% Zarrouk\u0026rsquo;s medium, while T2 involved a dilution of Zarrouk\u0026rsquo;s medium to 50%. T3 and T4 comprised combinations of 50% Zarrouk\u0026rsquo;s medium with 50% parboiled effluent and 50% RO wastewater, respectively. Each treatment was replicated thrice to ensure the reliability of the results. A two-liter formulated low-cost medium was prepared and inoculated with a 100 ml culture of Spirulina (\u003cem\u003eSpirulina subsalsa\u003c/em\u003e) at a 0.5 optical density, sourced from a two-week-old fresh culture. The cultivation setup was placed under 65% shade within a net house environment. Regular readings were recorded until a consistent optical density (OD) was achieved, signaling the maturity of the Spirulina biomass. Following this, the biomass was harvested, filtered using Whatman No. 42 filter paper, and subjected to oven drying at 60\u0026deg;C for a duration of 48 hours.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eAzolla Cultivation\u003c/h2\u003e \u003cp\u003eA complete randomized design with nine treatments (T1 - Kitchen wastewater\u0026thinsp;+\u0026thinsp;triple super phosphate (TSP), T2- Tap water\u0026thinsp;+\u0026thinsp;TSP, T3- RO wastewater\u0026thinsp;+\u0026thinsp;TSP, T4 - Kitchen wastewater\u0026thinsp;+\u0026thinsp;Cow dung, T5 - Tap water\u0026thinsp;+\u0026thinsp;Cow dung, T6 - RO wastewater\u0026thinsp;+\u0026thinsp;Cow dung, T7 -Kitchen wastewater, T8 - Tap water, T9 - RO wastewater) and triplicates was prepared for Azolla cultivation to ensure statistical significance and reproducibility. Tanks (30\u0026times;15 cm) were filled with a 10 cm depth of selected water. Eleven g of 2 days decomposed cow dung and 0.5 g triple super phosphate (TSP) was added with selected water based on decided treatments [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The setup was kept for 2 days to settle down the particles. 0.1 g of fresh Azolla biomass was crushed and added to each tank separately. The setup was kept under 65% shade net house condition. After 14 days biomass was harvested and oven dried at 60\u003csup\u003e0\u003c/sup\u003eC for 48 hours to reach the constant weight and dry weight was measured.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eAnalysis of Organic Sources and Plant Tissue\u003c/h2\u003e \u003cp\u003eOrganic sources were prepared through a sequence of chopping, washing, drying, grinding, and sieving through a 2 mm sieve. The powdered samples were stored in airtight conditions for nutrient analysis. The determination of total nitrogen, phosphorus, potassium, and total organic carbon content was carried out employing established laboratory standard methodologies such as total nitrogen (Kjeldhal method [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]), phosphorus (Vanadomolybdate method [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]) potassium [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], total organic carbon [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] and Calcium and magnesium were measured using an atomic absorption spectrophotometer.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eFormulation of LOF\u003c/h2\u003e \u003cp\u003eLOF was formulated by mixing 25% Banana pseudostem, 25% Palmyrah leaf, 25% Coconut leaf, 1% Spirulina, and 24% Azolla powders based on availability, nutrient analysis and C/N ration to achieve the level of N (1.47%), P (0.34%), K (1.12%) and C/N ratio (24.66) as equal or higher than that of the organic fertilizer, cattle manure N (1.42%), P (0.31%), K (1.21%) and C/N ratio (23.42).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eGermination and Vigor Test\u003c/h2\u003e \u003cp\u003eSeeds were soaked for 3 hours with 2% of seed weight in an organic source. Curry Chilli and Green Chilli seeds were placed in filter paper, while Water spinach and Sugargraze seeds were placed in sand beds [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Germinated seeds were counted at 2-day intervals. Seedling height was measured, and germination percentage and seedling vigor were calculated based on standard procedures. The percentage of germination was calculated by dividing the number of seeds germinated by the total number of seeds sown [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003ePercentage of germination = (Number of seeds germinated / Number of seeds sown) \u0026times; 100%\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eSeedling height (cm): Three seedlings were randomly uprooted from each replicate in each treatment and height was measured in centimeters. The Vigour index was estimated using the following formula (Sharma et al., 2014).\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eVigour index = (Seedling height (cm) \u0026times; Germination percentage)\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003ePot Experiment\u003c/h2\u003e \u003cp\u003eThe pot experiment focused on Sugargraze as the test crop and was organized in a complete randomized design (CRD) comprising seven distinct treatments denoted as T1 to T7. These treatments included T1 - Control, T2\u0026ndash;100% IN, T3\u0026ndash;50% IN, T4\u0026ndash;100% CM, T5\u0026ndash;50% CM, T6\u0026ndash;50% IN\u0026thinsp;+\u0026thinsp;50% LOF, T7\u0026ndash;50% CM\u0026thinsp;+\u0026thinsp;50% LOF (IN: Inorganic fertilizer based on department recommendation, CM: cattle manure, LOF: low-cost organic fertilizer). Black polythene bags were used as pots with sieved soil. Organic treatments received cattle manure and low-cost organic fertilizer, inorganic treatments received urea (150 Kg/ha), Muriate of potash - MOP (50 Kg/ha), and Triple super phosphate - TSP (100 Kg/ha) fertilizers and they were applied at the Department of Agriculture (DOA) Sri Lanka recommended rates.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eData Collection and Statistical Analysis\u003c/h2\u003e \u003cp\u003eData on growth and yield parameters were collected, and statistical analysis was performed using the Statistical Analytical System (University version) with Duncan's mean separation at P\u0026thinsp;=\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e"},{"header":"Result and Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eBiomass Yield of Spirulina\u003c/h2\u003e \u003cp\u003eThe efficiency of semi-mass culturing was measured in terms of biomass yield, expressed in dry weight per unit volume. In this study, treatment with 50% Zarrouk\u0026rsquo;s medium\u0026thinsp;+\u0026thinsp;50% parboiled effluent (T3) exhibited a significantly higher dry biomass yield compared to treatment with 100% Zarrouk\u0026rsquo;s medium (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). It was observed that treatments with 100% Zarrouk\u0026rsquo;s medium (T2) and 50% Zarrouk\u0026rsquo;s medium\u0026thinsp;+\u0026thinsp;50% RO wastewater (T4) did not exhibit significant differences in dry biomass. Notably, 50% Zarrouk\u0026rsquo;s medium\u0026thinsp;+\u0026thinsp;50% parboiled effluent (T3) gave the highest dry biomass (3.023 g/l) of Spirulina on the 30th day of cultivation because the parboiled effluent, rich in ammonia-nitrogen, may have contributed to the increase in biomass yield in T3. Parboiled rice effluent is known to contain high levels of organic matter, nitrogen, and phosphorus [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] while RO wastewater comprises both organic and inorganic components, including single-charged ions such as Na\u003csup\u003e+\u003c/sup\u003e and Cl\u003csup\u003e\u0026minus;\u003c/sup\u003e [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Treatment combinations involving parboiled effluent and RO wastewater provided the necessary nutrient levels for Spirulina growth, resulting in higher biomass yields. The utilization of 50% wastewater, such as parboiled effluent or RO wastewater, as a substitute for Zarrouk\u0026rsquo;s medium, was found to halve the production costs needed for medium because in the conventional process of Spirulina biomass production, cultivation utilizing chemical-based culture mediums accounts for 35% of the total production cost. Furthermore, the environmental impact during the cultivation stage is notably the most significant among all production stages, primarily due to the extensive use of chemicals and nutrients [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Additionally, treatments involving wastewater sources (T3 and T4) yielded higher biomass compared to T2 (50% Zarrouk\u0026rsquo;s medium alone), suggesting that wastewater serves as a nutrient supplement for Spirulina cultivation. Therefore, employing wastewater instead of clean water for Spirulina cultivation presents a viable solution to address water scarcity issues. According to Richmond (2004), the biomass production of Spirulina relies heavily on factors such as temperature and nutrient availability in the growing medium. The synthetic standard growing medium for spirulina cultivation is Zarrouk's medium, which is rich in essential elements such as sodium, potassium, calcium, magnesium, and traces of iron and manganese [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. However, the cost of Zarrouk's medium is relatively high, and it typically requires laboratory-grade clean water for preparation, contributing to operational expenses [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. In a previous study, Mukherjee et al., 2016 reported that microalgae cyanobacteria consortium was grown successfully in parboiled effluent and the biomass was used as fertilizer. In another study [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] rice mill wastewater in combination with sodium chloride was used to grow Spirulina. Usharani et al., 2012 used parboiled effluent in combination with sodium bicarbonate.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTreatments Scheduled for Spirulina and Azolla Semi - Mass Culturing, Recorded Data\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eSpirulina\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eAzolla\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003cp\u003eCombination\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003cp\u003eBiomass\u003c/p\u003e \u003cp\u003eYield\u003c/p\u003e \u003cp\u003e(g/l)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003cp\u003ecombination\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003cp\u003eBiomass\u003c/p\u003e \u003cp\u003eYield\u003c/p\u003e \u003cp\u003e(g/l)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100% Zm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.81\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eKw\u0026thinsp;+\u0026thinsp;TSP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.14\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50% Zm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.45\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTw\u0026thinsp;+\u0026thinsp;TSP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.21\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50% Zm\u0026thinsp;+\u0026thinsp;50% Parboiled effluent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.20\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRO\u0026thinsp;+\u0026thinsp;TSP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.97\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50% Zm\u0026thinsp;+\u0026thinsp;50% RO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eKw\u0026thinsp;+\u0026thinsp;Cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTw\u0026thinsp;+\u0026thinsp;Cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.09\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRO\u0026thinsp;+\u0026thinsp;Cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.14\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eKw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.60\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.57\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.64\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eThe same letters within columns are not statistically different by the DUNCAN at p\u0026thinsp;=\u0026thinsp;0.05\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e(Kw - Kitchen wastewater, Tw - Tap water, RO - Reverse osmosis wastewater, Cd - Cow dung, TSP - Triple super phosphate Zm - Zarrouk\u0026rsquo;s medium)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eAzolla Fresh and Dry Biomass Yield\u003c/h2\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e presents the dry biomass yield of Azolla. Among all treatments, RO wastewater\u0026thinsp;+\u0026thinsp;cow dung (T6) obtained the highest dry biomass yield and it was significantly higher than other treatments. Next to T6 (RO wastewater\u0026thinsp;+\u0026thinsp;cow dung) T4 (Kitchen wastewater\u0026thinsp;+\u0026thinsp;Cow dung) gave high dry biomass yield. The treatments that includes wastewater components shows higher or equal dry biomass yield compared to treatments that includes tap water (fresh water). Treatments combined with cow dung gave a high dry biomass yield compared with other treatments. The results of this study reveal the potential of utilizing wastewater as a viable growth medium for Azolla cultivation, a practice not commonly explored in traditional cultivation methods where Azolla is typically grown in well water, pond water, and freshwater except few studies where Azolla was used to remediate wastewater rich in ammonium and phosphorus [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] The significant increase in dry biomass yield observed in treatments incorporating wastewater components underscores the efficacy of waste utilization in enhancing Azolla growth.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eNutrient Analysis of Organic Sources\u003c/h2\u003e \u003cp\u003eFive different organic sources, cattle manure and LOF were analyzed. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e presents the primary nutrients (C, N, P, and K) and secondary nutrients (Mg and Ca) available in organic sources. Among the organic sources, Spirulina recorded the highest N content (115.03ppm), followed by Azolla, Coconut Leaf, Palmyrah Leaf, Banana Pseudostem, and Cattle Manure. Banana pseudostem gave the highest P (6.17ppm) content, while Spirulina has the lowest. Banana pseudostem also contains the highest K content (19.56ppm), whereas Coconut Leaf has the lowest. Palmyrah Leaf has the highest C (832.50 ppm) content, followed by Coconut Leaf, Banana Pseudostem, and Spirulina. Among secondary nutrients, Azolla recorded the highest amount of Ca (10861ppm) and Mg (1947.2ppm). The LOF is formulated to match the nutrient content of the prevailing organic source, in this case, cattle manure, in a proper ratio. By formulating LOF to have nutrient content equivalent to cattle manure, it ensures a consistent supply of nutrients for plant growth, utilizing locally available resources effectively. This approach promotes sustainable agriculture by utilizing organic waste materials as nutrient sources.\u003c/p\u003e \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eNutrient Content of Organic Sources\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 7.2482%;\"\u003e\n \u003cp\u003eOrganic Sources\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\" style=\"width: 13.7589%;\"\u003e\n \u003cp\u003ePrimary Nutrients\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\" style=\"width: 17.2974%;\"\u003e\n \u003cp\u003eSecondary Nutrients\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003eK\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003eMg\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003cp\u003e(ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eCoconut leaf\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e1.87\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e3.56\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e7.19\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e430\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003e901.6\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003e4099.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eSpirulina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e115.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e1.26\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e4.97\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e415\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003e1919.55\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003e2134.4\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eAzolla\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e32.43\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e1.06\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e14.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e317.5\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003e1947.2\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003e10861\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003ePalmyrah leaf\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e11.43\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e4.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e6.15\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e832.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003e1348.38\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003e3423.47\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eBanana pseudostem\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e7.00\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e6.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e19.56\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e330\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\n \u003cp\u003e1665.6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\n \u003cp\u003e184\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eCattle Manure\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e14.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e12.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e332.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 14.4877%;\"\u003e\n \u003cp\u003eLOF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.7951%;\"\u003e\n \u003cp\u003e14.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 4.6536%;\"\u003e\n \u003cp\u003e3.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.0926%;\"\u003e\n \u003cp\u003e11.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 5.8829%;\"\u003e\n \u003cp\u003e362.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 7.4633%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.8487%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"8\" style=\"width: 42.3997%;\"\u003eThe same letters within columns are not statistically different by the DUNCAN at p\u0026thinsp;=\u0026thinsp;0.05\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n\u003c/table\u003e\u003cp\u003eGermination Percentage and Vigour Index\u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e shows the effect of different treatments on germination percentage and vigor index. The germination test conducted for Curry Chilli, Water spinach, Green Chilli, and Sugargraze indicated that the utilization of Spirulina, Azolla, and LOF as soaking agents resulted in enhanced germination rates compared to the control group. Moreover, seedling vigor parameters exhibited notable improvements across all treatments when compared to the control (T5). Compared to other treatments T3 and T4 gave high germination percentages and vigor index; the production of growth substances and vitamins by the algae may be partly responsible for this [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Spirulina, Azolla, and Banana contain phytohormones like Gibberellins, Auxin, and Cytokinin itself [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Gibberellin can break the seed dormancy and increase the germination percentage [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e], which could have contributed to higher germination percentage and germination vigor recorded in all treatments than control. Consequently, the application of Spirulina, Azolla, and LOF demonstrated positive effects on both seed germination and vigor index, suggesting their viability as commercially and ecologically sustainable biostimulants.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of different treatments on germination percentage and germination vigor of selected seed\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eWater spinach\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eCurry Chilli\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eGreen Chilli\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eSugargraze\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eG.P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eV.I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eG.P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eV.I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eG.P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eV.I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG.P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eV.I\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1495.21 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e96%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e909.34 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e662.07 \u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e97%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2201.11 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e91%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1441.61 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e775.72 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e96%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e611.91 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e97%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2008.89 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e96%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1614.09 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e96%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e923.34 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e691.11 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e97%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2258.89 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e93%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1611.63 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e99%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e882.12 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e99%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e669.34 \u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2288.89 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e87%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1137.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e93%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e692.30 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e94%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e509.88 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e87%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e86.6667 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eT1 - Low-cost organic fertilizer, T2 - Dry Azolla powder, T3 - Dry Spirulina Powder, T4 - Live Spirulina Culture, T5 - Control (Distilled water) G.P - Germination Percentage, V.I \u0026ndash; Vigor Index\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003ePot Experiment - Sugargraze\u003c/h2\u003e \u003cp\u003ePlant Height\u003c/p\u003e \u003cp\u003eThe plant height at 2-week intervals is illustrated in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Almost during all weeks significantly higher plant height was observed in T2 (100% inorganic) and T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF) than other treatments. However, there were no significant height differences between T2 (100% inorganic) and T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF). During 10th and 12th weeks after seeding the height difference among treatments was similar. It is significant to note that plant height in T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) was significantly higher than that of T4 (100% organic) after 4th week.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePlant height - Sugargraze\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2nd week\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4th week\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6th week\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8th week\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10th week\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e12th week\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e36.50 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e48.50 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e75.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e102.00 \u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e121.00 \u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e41.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e120.00 \u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e172.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e191.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e210.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35.00 \u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e101.50 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e142.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e161.00 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e183.00 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31.00 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e104.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e147.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e181.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e193.50 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.75 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e40.50 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e62.00 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e141.00 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e150.50 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e41.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67.5 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e124.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e171.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e192.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e210.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34.50 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e111.00 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e157.50 \u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e186.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e195.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eT1 - Control, T2\u0026ndash;100% inorganic, T3\u0026ndash;50% inorganic, T4\u0026ndash;100% organic, T5\u0026ndash;50% organic, T6\u0026ndash;50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF, T7\u0026ndash;50% organic\u0026thinsp;+\u0026thinsp;50% LOF\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFresh and Dry Biomass Weight\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the fresh and dry biomass weight of plants with different treatments. The high fresh weight per plant was observed in T4 (100% organic) and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) while the lowest was in T1 (Control). Fresh weight of T4 (100% organic), T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF), and T7 (50% CM\u0026thinsp;+\u0026thinsp;50% LOF) were not statistically significant among each other.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe higher dry weight per plant was observed in T4 (100% organic) and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) while the lowest was in T1 (Control). There were no significant dry bioma\u0026aacute;ss differences among T2 (100% inorganic), T4 (100% organic), T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF), and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF).\u003c/p\u003e \u003cp\u003eNutrient Content (N, P, K) - Sugargraze\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the nutrient content in plants with different treatments. The high N uptake was recorded with T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF) and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) and it was significantly higher than other treatments. The lowest N uptake was noted in T1 (Control).The high P uptake was observed in T4 (100% organic) \u0026amp; T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF). P uptake in T4 (100% organic) \u0026amp; T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) were significantly higher than other treatments however there were no significant differences among them. The low P uptake was recorded with T1 (Control) and T3 (50% inorganic). K uptake in the plant was high in T4 (100% organic), T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF), and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF). In accordance with N, P, and K the sole organic treatments namely T4 (100% organic) \u0026amp; T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF), and IPNS T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF) had a higher % of nutrient content than that of T2 (100% inorganic) because of high nutrient leaching problem in 100% inorganic system even though it has high nutrient availability [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the pot experiment, treatments T2 (100% inorganic), T6 (50% inorganic\u0026thinsp;+\u0026thinsp;50% LOF), and T7 (50% organic\u0026thinsp;+\u0026thinsp;50% LOF) exhibited significantly higher values in crop growth parameters, dry biomass yield, and nutrient content in comparison to other treatments. Notably, there were no significant differences observed among these three treatments. Significantly higher growth and yield of Wheat and nutrient uptake in combined organic-inorganic treatments than sole inorganic treatment [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], better yield performance and quality of Mustard in integrated nutrient treatments [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] better performance of integrated nutrient management in different crops also reported by Fan et al., 2008 and Selim 2020. From an environmental and health perspective, the combination of 50% inorganic fertilizer with LOF (T6) or the sole organic combination with LOF (T7) emerged as viable alternatives to the 100% inorganic treatment (T2). The integrated nutrient management tool efficiently supplies plant nutrients, lowers costs, improves soil conditions, promotes a healthy environment, maintains nutrient balance, and manages agricultural waste safely [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. The findings of the present study underscores that substituting 50% of organic or inorganic fertilizer with LOF can yield comparable or higher performance in sugargraze cultivation.\u003c/p\u003e \u003c/div\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe research findings reveal the potential of Spirulina, Azolla, and LOF as bio-stimulants in enhancing seed germination and vigor across multiple crops. Spirulina semi mass culturing experiments revealed that incorporating wastewater sources significantly enhanced biomass yield, offering a cost-effective solution and addressing water scarcity issues. Azolla cultivation experiments emphasized the potential of utilizing kitchen and RO wastewater in combination with cow dung, to achieve comparable or higher biomass yields compared to tap water in combination with cow dung. Importantly, underutilized organic sources and nitrogen sources from wastewater were identified as valuable alternatives to conventional fertilizers, Spirulina serves as a promising alternative source for urea, Azolla addresses secondary nutrient deficiencies (Ca, Mg), Coconut leaf and Palmyrah leaf act as valuable sources of carbon, and Banana pseudostem offers an alternative source for triple superphosphate and muriate of potash. In the pot experiment of sugargraze, combinations involving LOF gave notable improvements in crop growth parameters, biomass yield, and nutrient content compared to conventional inorganic treatments. Particularly, substituting 50% of organic or inorganic fertilizer with LOF proved beneficial from both environmental and health standpoints. These organic sources demonstrate significant potential to replace inorganic fertilizers in agricultural practices. These findings collectively advocate for the adoption of environmentally sustainable practices in agriculture, highlighting the feasibility of integrating organic nutrient sources to enhance crop productivity while minimizing environmental impact and resource consumption.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe affirm that we did not receive any funds, grants, or additional support during the preparation of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no relevant financial or non-financial conflicts to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBoth authors contributed to the conception and design of the study. Material preparation, data collection, and analysis were conducted by Kasthuri Kajeevan. The initial draft of the manuscript was authored by Kasthuri Kajeevan and subsequently revised by Nalina Gnanaverajah. Both authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and analyzed during the current study are not publicly accessible due to commercialization purposes. However, they can be obtained from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eGomiero, T., Pimentel, D., Paoletti, M.G.: Environmental impact of different agricultural management practices: conventional vs. organic agriculture. CRC. Crit. Rev. Plant Sci. \u003cb\u003e30\u003c/b\u003e(1\u0026ndash;2), 95\u0026ndash;124 (2011)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSavci, S.: Investigation of effect of chemical fertilizers on environment. 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Arch. \u003cb\u003e3\u003c/b\u003e(6), 1327\u0026ndash;1341 (2012)\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Azolla, Crop productivity, Integrated plant nutrition system, Organic farming, Wastewater utilization, Spirulina","lastPublishedDoi":"10.21203/rs.3.rs-4411495/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4411495/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose:\u003c/strong\u003e This research investigated sustainable agricultural practices focusing on the utilization of wastewater and underutilized organic nutrient sources. The study explores the cultivation of nitrogen-rich sources such as Spirulina and Azolla using wastewater as a growth medium, aiming to reduce production costs and address water scarcity problems. Additionally, a low-cost organic fertilizer (LOF) was developed using underutilized resources like Palmyrah leaves, Coconut leaves, and Banana pseudostem, with Spirulina, and Azolla aiming to counteract the detrimental effects of inorganic fertilizers and contribute to the circular economy by converting waste into valuable agricultural inputs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Experimental trials assessed the efficacy of wastewater as a growth medium for Spirulina and Azolla, alongside evaluating the impact of organic nutrient sources on seed germination and seedling vigor. Pot experiments were conducted to assess biomass yield and crop growth parameters across various fertilizer treatments, including conventional organic, LOF, and inorganic methods.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Results reveal that treatments containing wastewater significantly enhanced the biomass yield of Spirulina and Azolla compared to standard methods, offering a cost-effective solution. The LOF enhanced the seed germination (5-15%), and seedling vigor (12-53%) of selected seeds compared to the control. Pot experiment revealed that the combination treatments of LOF gave improvements in crop growth parameters, biomass yield, and nutrient content of Sugargraze (Hybrid sweet sorghum) compared to conventional organic or inorganic treatments. Notably, substituting 50% of organic or inorganic fertilizer with LOF gave comparable or higher performance, highlighting the potential of LOF as an environmentally sustainable alternative.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e Overall, this study underscores the feasibility of integrating underutilized organic nutrient sources and wastewater utilization in sustainable agriculture.\u003c/p\u003e","manuscriptTitle":"Enhancing Sustainable Agriculture through Wastewater Utilization and Low-Cost Organic Fertilizer Production","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-12 06:56:11","doi":"10.21203/rs.3.rs-4411495/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"21c5c7a5-e768-46ea-bf0a-c05f566374fe","owner":[],"postedDate":"June 12th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-12T20:44:12+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-12 06:56:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4411495","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4411495","identity":"rs-4411495","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}