Study of remediation efficiency and biomass production using cyanobacteria and microalgae in secondary treated tannery wastewater

doi:10.21203/rs.3.rs-2991386/v1

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Study of remediation efficiency and biomass production using cyanobacteria and microalgae in secondary treated tannery wastewater

https://doi.org/10.21203/rs.3.rs-2991386/v1

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Secondary treated tannery wastewater (STTWW) is high in COD with value ranges from 264 mg/L to 275 mg/L also, the total suspended solids value was 164. The total nitrogen calculated was 33.2 mg/L. Cyanobacteria and microalgae are efficient bioremediators as well as significant biomass producers with loads of bimolecular content such as carbohydrate, lipid, protein which can be used for the production of value added products(VAPs). Simple tubular photobioreactors were made for mass culturing both cyanobacteria and microalgae. Cyanobacteria had higher amount of protein (0.58 mg/L) at 100% STTWW at pH 7 and at light intensity of 1000 lumens, carbohydrate (1801.4 mg/L), chlorophyll (6.6303 mg/L) 100% STTWW at pH 7 and at light intensity of 1000 lumens. Lipid was higher at 40% STTWW when compared to microalgae the remediation efficiency of cyanobacteria was better than microalgae. Cyanobacteria reduced COD reduced upto 94 % Overall this study depicts that cyanobacteria is more efficient in bioremediation and biomass production.

Sustainable remediation of industrial wastewater is a pressing environmental concern, particularly in the case of tannery effluent wastewater which contains high concentrations of organic, inorganic compounds (Chandra et al., 2011). Tannery industries are prominent in words economy providing employment opportunities and generating income. India became one of the leading players in the leather industry with the higher availability of technology for processing leather, water resources, and trained laborers (Suthanthararajan et al., 2022). The export of this sector reached 2 billion in the 2014 data report, which is 10% of the world’s share, 100 million pairs of shoe uppers, 16 million pieces of garments of leather, 52 million pairs of hand gloves, and 909 million footwear, being inevitable part (Ricky et al.,2022). Tannery industries undergo various steps involved in tanning to produce leather from raw hide which is stable and non-destructive form. The major steps in the process of tanning are soaking which introduce high amount of organic matter, dissolved salts and suspended solids. Liming is usually done by using calcium hydroxide which incorporates organic matter, fats. Dehairing process incorporates chemicals used for dehairing resulting in increased suspended solids and organic content (Chandra et al., 2011). The fleshing generates fats, organic content and suspended solids, deliming and batting contributes residual lime, organic matter, enzymes, and fats into generated wastewater. The main step tanning increases the levels of chromium salts (in case of chrome tanning), organic matter and dissolved salts. After tanning neutralization and dyeing generated wastewater introduced to chemicals, such as dyes and acids, suspended solids and organic matter. From above mention process the wastewater produced in tanneries become highly complex and rich in organic contents, dissolved salts, suspended solids and potentially harmful chemicals ammonia, hydrogen sulfide, volatile hydrocarbons, amines, aldehydes chromium, synthetic tannins, oils, resins, biocides, and detergents are emitted to the atmosphere from tannery plants as effluent (Islam et al., 2014). The treatment of tannery produced wastewater becomes very significant before releasing to the open water bodies and environment as it may poses harmful effects for environment as well as to humans.

To eliminate the problem associated with wastewater discharge wastewater treatment process such as physical (filtration, adsorption), chemical (coagulation/ flocculation) advanced treatment methods (membrane filtration, activated carbon adsorption, AOPs) and biological process (microbial treatment), phytoremediation, constructed wetlands are employed. All mentioned techniques have their own drawbacks and lack in potential (Cheng et al., 2021). Chemical methods introduce chemical to wastewater and produce large amount of sludge which become difficult to dispose. Biological processes (bacteria, fungi, yeast) are non-rewarding as simple bioremediation is needs high cost for rectors setting ups and produces secondary pollutants as sludge (Verma et al., 2019). The sludge becomes difficult to process and handle. Using organism of value like cyanobacteria, which is photosynthetic Cyanobacteria are a group of photosynthetic organisms that can easily survive with the minimum requirement of light, carbon dioxide, and water. Being phototrophic in nature and grow in a variety of locations. They fix nitrogen and produce some bioactive compounds, which improve the nutritional status of the soil, promote the growth of crops, and protect them from plant pathogens. Cyanobacteria have the ability to change their modes of nutrition: biogeochemical cycles by making essential transformations to the organic compounds (Yadav et al., 2021) and microalgae are a diverse group of prokaryotic and eukaryotic photosynthetic microorganism that can grow rapidly due to their simple structure (Rossi and Phillips, 2015). The microalgae have advantages over the cultivated plants such as faster growth, yield of high volume of oils and possibility of the use of any kind of water for their culture, they are tolerant to high concentration of salts Both cyanobacteria and microalgae are not only having the remediation quality but, also can resolve problem of secondary pollutant. The recovered valued biomass after treatment of tannery wastewater (TWW) can be utilized for production of various value-added product (VAPs) by analyzing the biomolecule composition (Chandra et al., 2018) The remediation of TWW with simultaneous production of value-added products is an initiative towards the development of a cost effective and green technology providing the additional economic advantage such as saving of chemicals like media preparation, reduced capital and operational cost with environmental benefits, TWW is highly rich in nitrogen, phosphorus, sulphur and some metal ions and serve as nutrient source for the effective microalgal growth (Yadav et al., 2021).

Photosynthetic microalgae use atmosphere CO₂ as carbon source and liberated O₂ is utilized by bacteria for their growth as well as in reduction of pollutants load (if microalgae and bacteria both are used together in bioremediation). But, heterotrophic microalgae use organic carbon as soluble carbonates directly or CO₂ as carbonates fixed by carbo-anhydrase enzyme. Also, the CO₂ produced by bacteria through respiration is absorbed as soluble carbonates by microalgae (Satyanarayana et al., 2011).

According to a reported data given by Sadvakasova et al., 2021, the use of biomass of cyanobacteria which actively accumulate lipid in the production of biofuel, recent studies have shown that the production of lipids by the Synechocystis PCC6803 strain was 12.5% (Maheshwari et al. 2018). The data on a sufficiently large number of lipids in the cells of cyanobacteria Oscillatoria sp. (31.9%), Synechococcus sp. (30.6%), Croococcidiopsis sp. (22.7%), Leptolyngbya sp. (21.15%), Limnothrix sp. (20.73%) are presented (Hossain et al. 2020). Palanisamy et al. (2021) established a high lipid accumulation of up to 34.5% for Spirulina phlatensis, which was increased to 35.8% after light optimization. Han et al. (2016) optimized the Anabaena variabilis strain for lipid production, achieving its productivity of up to 46.9%. Yalcin and Hoffmann (2020) cultivated the freshwater cyanobacteria Dolichospermum affine strain under starvation and limitation conditions, using six different concentrations of nitrate and phosphate in order to study fatty acid profile. Lipid content of the strain was 10.67% dry weight. Another related work was hold with Nagappan et al. (2020) research group. As previously mentioned, Bolatkhan et al. (2020) investigated several cyanobacterial strains for lipid production that grown in handmade photobioreactor and the result showed unicellular cyanobacteria Cyanobacterium sp. IPPAS B-1200 contains 16.1% lipid from dry weight biomass. As reported by Blažina et al. (2019) Synechococcus sp. MK568070 strain investigated as a high productive for biodiesel production and lipid content was higher from previously mentioned strains − 21.4% dry weight.

Co-cultivation of microalgae and cyanobacteria has been reported by various researchers to explore the symbiotic relationship between microalgae (chlorophytes) and cyanobacteria. B. braunii and N. muscorum co-culturing resulted in 38% enhancement in biomass content and 27% increase in lipid content due to the presence of modifed secondssary metabolites developed during co-culturing, Microalgal biomass produced during wastewater treatment can further be used for the production of pigments, nutraceuticals, biofuels, biomolecules, biofertilizers, and feed additives cyanobacteria biomass include several processes: biodiesel (transesterifcation of biomass); biogas (anaerobic digestion); bioethanol (fermentation); and biohydrogen (photosynthetic processes or dark fermentation). C. vulgaris cultivation in seafood wastewater effluent showed high lipid content of 32.15% for biodiesel production (Nguyen et al. 2019). C. sorokiniana cultivation in raw urban wastewater showed 97% nutrient removal, 44% total organic carbon removal, biomass productivity of 1.13 g L^− 1, and 31% dry weight lipid content (Arora et al. 2020). This research is focused on the nutrient recovery from secondary treated tannery wastewater by analyzing the physicochemical and biomass biomolecule analysis for further production of value-added products (VAPs)

2.1. Collection of Secondary treated tannery wastewater and sludge samples

The secondary treated tannery wastewater and sludge were collected in sterilized container from the Common Effluent Treatment Plant (CETP) of tannery industries located at Unnao, (26.48° N latitude, 80.43° E longitude), (U.P), India. The collected wastewater samples were brought to laboratory and stored at 4°C for physicochemical analysis.

2.1.1. Characteristics and physiochemical analysis of secondary treated tannery wastewater

Physiochemical analysis of the tannery effluents was performed following the standard methods by APHA (2017). The parameters analyses were color, conductivity, pH, TDS, BOD, COD and total nitrogen, EC, Cl⁻, Mg²⁺, Ca²⁺, HCO³⁻, CO³⁻, TSS, TN, Phosphorus.

2.2. Isolation and identification of pre-existing cyanobacteria and microalgae lab culture

A total two samples were collected from the CETP, Unnao Uttar Pradesh. Samples were bought to Lab with minimal contamination in clean, sterile sample collection bottles at a temperature of 4° C and kept in sterile condition for the analysis of the samples (Chittapun etal., 2017).

2.2.1. Isolation of cyanobacteria and microalgae

The cyanobacteria and microalgae isolated in this study is from the laboratory itself (Laboratory of Bioremediation and Metagenomics Research of the Department of Environmental Microbiology, BBAU, Lucknow) The collected samples were enriched initially in BG-11. Cyanobacterial and Microalgae samples were processed under aseptic conditions. 10 ml ofBG11 broth was prepared and then 1ml sample was inoculated into the BG11 broth. After inoculation, the cultures tube was kept for 20 days at 25–28°C and the cultures were illuminated with light intensity 1000 lumen at the surface of vessels or tube for better growth and then repeat for 3 times. The purity of the culture was monitored by regular observation under a microscope and the morphological characterization of the culture (Ferrish et al., 2015).

2.2.2. Identification of cyanobacteria and microalgae

Further the sample were examined under an electron microscope for the presence of cyanobacteria and microalgae. Identification of cyanobacteria and microalgae was conducted with aid of taxonomic keys and literature (Wisniewska et al., 2020).

2.3. Experimental setup

A methodology is presented for designing tubular photobioreactor consisted of recycled Polyethylene Terephthalate (PET) bottles, pipe, T-connector, LED light (1000 lumen), aquarium motor. The design methods effectively remediate secondary treated tannery wastewater (STTWW) and also produce biomass. The reactor was located in the chamber with white background to improve reflectance (Iqbal et al, 1993).

2.3.1. Construction of Photobioreactor

This research works aims to fabricate an optimized up-scaled photobioreactor using PET bottles to harvest the cyanobacteria and microalgae using STTWW and extract biomass as well as remediate the STTWW. Microalgae and cyanobacteria can be grown in photobioreactors (PBRs), where parameters like lighting, gas exchange, and mixing can be systematically controlled like for air circulation system, two holes were made in the cap where first one is connected to pipe i.e., air inlet pipe and other left open for proper delivery of oxygen.

The aquarium motor was used for the aeration system in the bottles filled with the concentration of autoclavable STTWW (strains used the nutrients in the wastewater to grow and produce oxygen) and tap water (shown in table) for the connection of the pipe the T-connector were used light source for photosynthesis process i.e., 1000 lumen. In the concentration pre-prepared inoculum with 1% (v/v) of cyanobacteria and microalgae strain were added. After exposing cyanobacteria and microalgae in light and oxygen then evaluate the productivity and efficiency of photobioreactor system in terms of biomass production, wastewater treatment and resource recovery (Skjånes et al., 2016). Experimental setup is shown in Table 1.

2.3.2. Mass culturing of Cyanobacteria using constructed photobioreactor

Mass culturing of cyanobacteria using constructed photobioreactor designed mainly the experiment uses different concentrations of STTWW (autoclave) and tap water(autoclave) in separate bottles. Bottle inoculated with 1% v/v of pre-prepared cyanobacterial strain in sterilized condition and kept in the design photobioreactor for 15 days under 1000 lumen light source. The cyanobacteria grow by using the nutrients present in then STTWW and produce biomass, which remediate the wastewater. After 15 days of incubation, the biomass is collected and dried in open atmosphere, while the remediated water is stored 4°C for further physiochemical analysis (Pruvost et al., 2016).

2.3.3. Mass culturing of Microalgae using constructed photobioreactor

Mass culturing of microalgae developed by taken 1% v/v of pre-prepared inoculum and inoculates in bottle with same concentration of STTWW and tap water in sterile condition (laminar air flow). The microalgae started to grow under the duration of 15days. The microalgae adopted the atmosphere and show growth in each concentration of bottles under the 1000 lumen of light. After 15 days, separate the microalgae biomass and remediated water through membrane filter paper and collect and dry into air. The remediated water stored into recyclable bottles at 4° C into freeze for physicochemical analysis (Pruvost et al.,2016)

2.2.4. Development of cyanobacteria and microalgae consortia for mass culturing using constructed photobioreactor

Mass culturing of cyanobacteria using constructed photobioreactor designed mainly the experiment employs different concentrations of STTWW (autoclave) and tap water(autoclave) in separate bottles. Bottle inoculated with1% v/v of pre-prepared inoculum of eachcyanobacterial and microalgal strainand kept in design photobioreactor for 15 days under 1000 lumen light source.Based on the observed performance of the photobioreactor system, further refine the consortium composition, growth circumstances, and operating parameters.

2.5. Analysis of produced biomass of cyanobacteria and microalgae (carb, lipid, protein, chlorophyll, biomass growth)

2.5.1. Carbohydrate estimation

Principle- This test can be used to access all carbs. In this test, furfural is reacted when carbohydrates react with conc. sulfuric acid to dehydrate them. This furfural interacts with anthrone to produce a blue-green coloured complex (Chen et al, 2013).

Using glucose as standard calibration

For the procedure take 1 ml of sample and place it in test tube add 1 ml of distilled water in another tube as a control then add 2 ml of anthrone reagent to all the test tube shake thoroughly the contents of the test tube and check out the color at 630 absorbance.

2.5.2.Lipid estimation

Principle: The technique is based on the idea that unsaturated lipids react with sulfuric acid and carbonium ion, vanillin reacts with phosphoric acid to produce an aromatic phosphate ester, and the carbonium ion reacts with the activated carbonyl group of phospho-vanillin to produce a charged, colored complex that is stabilized by resonance and absorbs light maximally at about 530 nm to produce (Byreddy et al., 2016). H₂SO₄ and phosphor- vanilline reagent: take 0.06gm vanilline add in 1ml of alcohol, 9ml of distilled water and 40ml of H₃PO₄(Orthophosphoric acid).

Procedure

In this technique oleic acid used as standard diluted with H₂SO₄ to reach 1 mg.ml ^− 1 then take 0.5 ml of sample in test tube add 2 ml of H₂SO₄ and boil the test tube for 10 minute, cool it for 5 minute and add 5 ml of phospho-vanillin reagent incubate it for 20 min at RT, then observe at absorbance at 530 nm.

2.5.3. Protein estimation

Folin Ciacateau phosphomolybdicphospho tungstic acid is reduced to heteropolymolybdenum blue by the copper-catalysed oxidation of aromatic acid in the Lowry method of protein concentration determination. This reduction occurs under alkaline conditions and is the basis for the method. Because the Lowry technique is sensitive to pH variations, the test solution's pH should be kept between 10 and 10.5(Waterborg, 2019). 2% Na₂CO₃ in 0.1N NaOH, 1% NaK Tartrate in H₂O, 0.5% CuSO₄.5H₂O in H₂O For this 1 ml of test tube with distilled water serve as blank, add 0.5 ml of sample and405 ml of Reagent 1 and incubate it for 10 minute after incubation add 0.5 ml of Reagent 2 and again incubate it for 30 minute, measure the absorbance at 660nm.

2.5.4. Chlorophyll estimation

1 ml of sample in Eppendorf and centrifuge at 4000 rpm for 10 minute discard the pallet and store the left supernatant in the refrigerator at 4° C for 24 hours then take the left supernatant and add 1 ml of acetone and centrifuge it on 4000 rpm for 15 minute. Further take the absorbance at 630nm, 645nm, 663nm, and 750nm after this calculate the total chlorophyll by the given formula (Jaiswal et al. 2018).

Chlorophyll a = 11.64\(\times\)(663nm-750nm) -2.16× (645nm-750nm) + 0.1 (630nm-750nm)\(\times\) Volume of sample

3.1. Characteristics and physicochemical analysis of secondary treated tannery wastewater

The secondary treated tannery wastewater was light brow in color with suspended solids and slightly pungent smell the analysis was given in table

3.2. Isolation and identification of cyanobacteria and microalgae for biomass production

The isolation of microalgae and cyanobacteria from the sludge done by take a sample from sludge. Firstly, 120 ml BG11 broth prepared by using BG11 media and A5 mix component with distilled water. 10 ml broth transfer in each culture tube in sterile condition using laminar air flow. In each culture tube 1ml inoculum transfer from the sample into the media and transferred inoculum in 2 culture tubes so prepared in twice. The media contains BG11 and A5 mix components (Yang et al. 2015).

The isolation of pure culture was done by using BG11 agar media. Prepare 100 ml of media for 4 petri dish plates and autoclave media at 15 psi pressure and 121̊ C temperature for 15 minutes. Taking 1ml sample from culture tube for spreading on agar plate media and kept in photobioreactor under illumination and observed colonies after 10–15 days. These colonies used as inoculum for streaking on BG11 agar plate media and observed result the pure colonies shows the growth on media. Using of pure culture for the remediation and biomass production. Prepare 120 ml BG11 broth in 250 ml conical flask and autoclaved at temprture121̊ C at 15 psi for 15 minutes. Take small amount of inoculum and inoculate into conical flask. Put for growth into photobioreactor (Wei et al., 2008).

Morphological identification and characterization is done by compound light microscope by using standard method. The growth observed by naked eyes. The inoculated 3and 4 sample is cyanobacteria because they appeared dark green in colour and other 1 and 2 sample are microalgae because it appeared light green in colour. The cyanobacteria shows dark green colour during the growth into the environment (Desikachary, 1959 and Hossain et al., 2016).

3.3. Bioremediation efficiency and biomass analysis

3.3.1. Cyanobacterial remediation efficiency and biomass analysis

Cyanobacteria remediated secondary treated tannery wastewater pH was observe after remediation pH 7.4 to 7.1. The cyanobacteria efficiently remediate chemical oxygen demand (COD) that observed 94.31%. There chlorine, calcium (Ca²⁺) and magnesium (Mg²⁺) remediate by cyanobacterial strain is 92.45%. Bicarbonate (HCO₃²⁻) observed after remediation efficiency 74.35% and the total nitrogen (TN) reduced by cyanobacteria 52.53%. The cyanobacteria show the high efficiency to remediate secondary treated tannery wastewater.

3.3.2. Microalgae remediation efficiency and biomass analysis

The remediation of STTWW by the microalgae shows less efficiency than cyanobacteria. The pH of STTWW reduces 7.5 to 7.1 after the treatment. Other parameter such as COD and total nitrogen (TN) detected value 89.09% and 31.32%. In treated STTWW with microalgae shows the efficiency of chlorine and bicarbonate is less as 21.05% and 40.27%. The reduction efficiency of calcium and magnesium shows 62.18% and 32.99%. It is also less that cyanobacteria remediation efficiency.

3.4. Comparative analysis

3.4.1. Comparative analysis of remediation efficiency of treated STTWW between cyanobacteria and microalgae

The comparative analysis shows that cyanobacteria are more efficient to remediate STTWW than microalgae (as shown in table). It analyzed that higher efficiency remediation of cyanobacteria strain value of COD, chlorine, calcium, magnesium, bicarbonate, carbonate and total nitrogen than microalgae. In cyanobacterial remediated STTWW, the parameter COD, chlorine and magnesium shown higher reduction efficiency as 94.31%, 92.45% and 97.21% instead of microalgae reduction efficiency are 89.09%, 21.05% and 32.99%. So it observed that nutrient utilization capacity of the cyanobacteria is greater than microalgae.

3.4.2. Comparative analysis of biomass between cyanobacteria and microalgae

The comparative analysis of biomass shows that higher amount of biomolecule found in cyanobacteria than microalgae. In cyanobacterial biomass, the amount of lipid, carbohydrate and chlorophyll is higher than in microalgae. The cyanobacteria is in higher amount in lipid 2.0241 mg/L present in 40% concentration of STTWW instead of microalgae lipid content 0.0171 mg/L; the amount of carbohydrate in cyanobacteria is 1801.4 mg/L in 100% concentration of STTWW but the amount of carbohydrate in microalgae 126.85 mg/L and the chlorophyll content of cyanobacteria is 6.6303 mg/L analyzed rather than the microalgae 0.2920 mg/L in 100% concentration of STTWW. The content of protein is higher value 0.580 mg/L in cyanobacteria rather than the content of protein 0.0591 mg/L present in 100% concentration of STTWW treated with microalgae. The biomass analysis show reduced value of cyanobacterial biomass than microalgae because the found amount of biomass 0.26 mg/L of cyanobacteria instead of microalgae amount 0.7105 mg/L in 100% concentration of STTWW

In conclusion, tannery wastewater can be remediated through the use of cyanobacteria and microalgae, are able to remove pollutants from wastewater, converting them into biomass, additionally cyanobacteria and microalgae mass cultured using photobioreactor and recover biomass. The recovery of biomass from STTWW using cyanobacteria and microalgae is an efficient approach for sustainable wastewater treatment and valued biomass production. The recovered biomass by cyanobacteria and microalgae harvested and use for the various purposes such as animal feed, biofuel, and biofertilizer, dietary supplements, cosmetics and also in nutraceutical industries.

The biomolecule like protein, lipid, carbohydrate, and chlorophylls were analyses for both cyanobacteria and microalgae as the result shown and we conclude that the cyanobacteria have more protein, lipid and carbohydrate content in comparison with microalgae. The studied cyanobacteria strains are promising protein, carbohydrate, lipid and chlorophyll as shown high content.

The study of cyanobacteria and microalgae interaction with secondary treated tannery wastewater reduce pH, colour intensity and the concentration of the chemical oxygen demand (COD), chlorine, calcium, magnesium, bicarbonate, total nitrogen (TN) of secondary treated tannery wastewater after 15 days. After remediation the concentration revealed by cyanobacteria 94.31%, 92.45%, 72.28%, 97.21%, 74.35%, 52.53% and by microalgae 89.09%, 21.05%, 62.18%, 32.99%, 40.27% and 31.32% respectively. The result shows that cyanobacteria strain indicted that better efficiency to remediate and use nutrient form secondary treated tannery wastewater.

The benefit of this study is to treat tannery wastewater is much less operating cost and eco-friendly process. The new biological-based technologies can supplement conventional treatment approaches, but may complement them especially secondary treated tannery wastewater.

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgement

Ms. Anuradha Devi is highly thankful to the ‘‘University Grants Commission’’ (UGC), New Delhi, India for providing the financial support for her research work.

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Table 1

Experimental design
CONCENTRATION 2(L)	TAP WATER (ml)	STTWW (ml)
20%	1600	400
40%	1200	800
60%	800	1200
80%	400	1600
100%	0	2000

Table 2

Physicochemical analysis of cyanobacteria (*Oscillatoria* sp.) and microalgae (Chlorella sorokiniana NMLX)
Physicochemical parameters	Control	Cyanobacteria	Reduction (%)	Control	Microalgae	Reduction (%)
pH	7.4	7.1	7.1	7.5	7.1	7.1
EC (mS/cm)	20.4		-	21.4		-
COD (mg/l)	264	15	94%	275	30	89.09%
Cl⁻ (mg/l)	53	4	52.92%	57	45	21.05%
Ca²⁺ (mg/l)	629.34	174.4	62.40%	619.34	234.2	62.18%
Mg²⁺ (mg/l)	842.1	23.42	84.20%	847.1	567.6	32.99
HCO³⁻ (mg/l)	53.26	13.66	53%	57.26	34.2	40.27
CO₃²⁻ (mg/l)	0.00	0.00	-	0.00	0.00	-
TSS (mg/l)	146	167	-	136	156	-
TN (mg/l)	33.2	15.76	32.72%	33.2	22.8	31.32%
P (mg/l)	2.65	-	-	2.65	1.79	-
*pH: Potential of hydrogen, EC: Electrical conductivity, COD: Chemical oxygen demand, Cl⁻: Chlorine, Ca²⁺: Calcium, Mg²⁺: Magnesium, HCO³: Bicarbonate, CO₃²⁻: Carbonate ion, TSS: Total solid substrate, TN: Total nitrogen, P: Phosphorus

Table 3

Obtained concentration of biomolecule cyanobacteria and microalgae
		20%	40%	60%	80%	100%
	Protein	126.582	110.548	178.340	414.064	580.028
	Lipid	297.87	205.455	239.886	454.32	312.686
(Oscillatoria sp.)	Carbohydrate	631.437	576.645	1519.563	1573.313	1801.438
	Biomass	0.1705	0.0945	0.0575	0.07	0.026
	Protein	12.687	32.068	22.675	44.444	59.072
	Lipid	30.46	17.676	25.897	47.764	77.852
(Chlorella sorokiniana NMLX)	Carbohydrate	397.062	436.229	254.770	279.562	126.854
	Biomass	0.41	0.487	0.315	0.4245	0.7105
	Chlorophyll	0.1818	0.18992	0.51502	0.47164	0.29195

No competing interests reported.

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Study of remediation efficiency and biomass production using cyanobacteria and microalgae in secondary treated tannery wastewater

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Abstract

Figures

1. Introduction

2. Material and method

2.1. Collection of Secondary treated tannery wastewater and sludge samples

2.1.1. Characteristics and physiochemical analysis of secondary treated tannery wastewater

2.2. Isolation and identification of pre-existing cyanobacteria and microalgae lab culture

2.2.1. Isolation of cyanobacteria and microalgae

2.2.2. Identification of cyanobacteria and microalgae

2.3. Experimental setup

2.3.1. Construction of Photobioreactor

2.3.2. Mass culturing of Cyanobacteria using constructed photobioreactor

2.3.3. Mass culturing of Microalgae using constructed photobioreactor

2.2.4. Development of cyanobacteria and microalgae consortia for mass culturing using constructed photobioreactor

2.5. Analysis of produced biomass of cyanobacteria and microalgae (carb, lipid, protein, chlorophyll, biomass growth)

2.5.1. Carbohydrate estimation

2.5.2.Lipid estimation

2.5.3. Protein estimation

2.5.4. Chlorophyll estimation

3. Result discussion

3.1. Characteristics and physicochemical analysis of secondary treated tannery wastewater

3.2. Isolation and identification of cyanobacteria and microalgae for biomass production

3.3. Bioremediation efficiency and biomass analysis

3.3.1. Cyanobacterial remediation efficiency and biomass analysis

3.3.2. Microalgae remediation efficiency and biomass analysis

3.4. Comparative analysis

3.4.1. Comparative analysis of remediation efficiency of treated STTWW between cyanobacteria and microalgae

3.4.2. Comparative analysis of biomass between cyanobacteria and microalgae

Conclusion

Declarations

References

Tables

Additional Declarations

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