The Comparison of Physiological Parameters of seven different Cyanobacterial strains

The Comparison of Physiological Parameters of seven different Cyanobacterial strains | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 25 July 2025 V1 Latest version Share on The Comparison of Physiological Parameters of seven different Cyanobacterial strains Authors : Anjali Singh 0009-0001-1473-7412 , Kamakshi [email protected] , and Devendra Kumar Authors Info & Affiliations https://doi.org/10.22541/au.175345758.85764042/v1 244 views 139 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract The diverse spectrum of pigments possessed by cyanobacteria, which are easily recognized by their characteristic blue-green colouring, enables them to exhibit a multitude of colors and hence adapt and flourish in a wide range of biological habitats. This study compares the physiological and physiochemical characteristics of isolated cyanobacteria strains procured from the Department of Microbiology, Chaudhary Charan Singh University (CCSU), Meerut. Physiological factors that were investigated include growth rates and pigment composition. These properties were assessed using spectrophotometry, growth curves, and standardized biochemical assays. The results indicated significant variations in the physiological traits of the cyanobacterial strains. Seven cyanobacterial strains were examined to analyze physiological parameters. The identified strains include species of Nostoc, Anabaena, Westiellopsis, Oscillatoria, Tolypothrix, Calothrix, and Phormidium . These strains were cultivated in BG-11 medium, and their pigment levels were analyzed at 7 th , 14 th , 21 st , and 28 th intervals throughout incubation. Distinguishable differences were seen among the different strains of these pigments. Maximum Chlorophyll, Carotenoid, Phycocyanin (PC), Protein, and Carbohydrate content was observed in Anabaena sp. on the 14 th day of incubation, whereas Tolypothrix sp. showed a maximum amount of Allophycocyanin (APC) and Phycoerythrin (PE). This thorough comparison of physiological data sheds light on cyanobacteria’s adaptation mechanisms and possible uses in biotechnology. Comprehending the varied physiological characteristics of these strains may have consequences for bioremediation, environmental management, and the creation of innovative biotechnological instruments across multiple sectors. Introduction Cyanobacteria are a substantial group of structurally intricate and ecologically important gram-negative prokaryotes, displaying diverse nutritional capabilities (Sánchez-Baracaldo et al. 2022). Cyanobacteria inhabit nearly every ecological niche on Earth, encompassing freshwater and saltwater environments, rice fields, geothermal springs, arid deserts, and polar regions (Bataeva et al. 2024). The ability of certain cyanobacteria to fix atmospheric nitrogen is a crucial biological function of economic significance (Nawaz et al.2025). Cyanobacteria exist both independently and in symbiotic partnerships with many species of the plant kingdom, including Gymnosperms, Pteridophytes, and Bryophytes (Chauhan et al. 2023). Yet their relationships with crop plants remain underexplored. They exhibit significant adaptability to diverse environmental conditions and have been extensively utilized as bio-inoculants to augment soil fertility, enhance soil structure, and improve crop yields, particularly in rice (Ríos-Ruiz 2025). The paddy-field ecosystem constitutes a distinctive aquatic-terrestrial habitat that fosters cyanobacteria’s growth and nitrogen fixation, satisfying their needs for water, increased temperatures, light, and nutrient availability. Cyanobacteria lead to organic matter, synthesize and release amino acids, vitamins, and auxins, decrease the oxidizable matter content of the soil, supply O 2 to the submerged rhizosphere, mitigate salinity, regulate pH, solubilize phosphates, and enhance fertilizer efficiency in crop plants (Murugesan et al. 2025). Many studies have documented the physiological and biochemical profile of one or several cyanobacterial strains under different environmental or stress conditions. El-Sheekh et al. (2005) examined chlorophyll and carotenoid levels in Nostoc muscorum under various nutritional contexts. Mouga et al. (2024) concentrated on growing the cyanobacterial strain Nostoc sp. 136 under regulated laboratory settings. Though these studies shed light on the metabolic capacity of certain strains, Comparative data on the physiological parameters of various cyanobacterial strains grown under the same laboratory settings are scant. A methodical comparison of several strains is required to find those with better metabolic and physiological profiles. The current study sought to acquire cyanobacterial isolates from selected paddy fields of western Uttar Pradesh that can produce metabolites such as proteins, pigments, and carbohydrates, which are valuable substances with potential applications in the food, pharmaceutical, and cosmetics industries. The present study aims to address these research gaps by conducting a comparative analysis of selected cyanobacterial strains isolated from paddy fields and exploring their physiological parameters. These traits play significant roles in determining their photosynthetic activity, stress tolerance, and metabolic processes, which are of substantial concern as used in biofertilizers, extraction of bioactive metabolites, and environmental remediation (Ansabayeva et al. 2025). The study emphasizes the importance of selecting effective strains based on their multifaceted growth-promoting activities, beyond their established role as nitrogen-fixers. Materials and Methods Culture selection Seven cyanobacterial strains specifically isolated from the selected paddy fields of Meerut, Muzaffarnagar, and Haridwar regions were procured from the algal culture collection of the Department of Microbiology, CCSU, Meerut, Uttar Pradesh. 250001 (Table 1). jabbrv-ltwa-all.ldf jabbrv-ltwa-en.ldf 1 Nostoc sp. Pabarsa, Meerut AS1 2 Anabaena sp. Jani Khurd, Meerut AS2 3 Westiellopsis sp. Mawana, Meerut AS3 4 Oscillatoria sp. Khatauli, Muzaffarnagar AS4 5 Tolypothrix sp. Budhana, Muzaffarnagar AS5 6 Calothrix sp. Laskar, Haridwar AS6 7 Phormidium sp. Khanpur, Haridwar AS7 jabbrv-ltwa-all.ldf jabbrv-ltwa-en.ldf Table 1 Selected Cyanobacterial strains and their designation used in the text (AS1-AS7) Isolation and identification of Cyanobacterial strains Cyanobacterial strains were cultured in a modified BG-11 medium at 28 ± 2°C, under controlled temperature at 16/8 hour light/dark cycle with a light intensity of 55 µmol photon m -2 s -1 and pH of 7.5 to optimize culture development (Fig. 1). After the 14 th day of the incubation period, cultures were streaked on the agar medium to isolate distinct colonies, which were subsequently transferred to 50 ml flasks containing 20 ml of BG-11 medium. To ensure the purity of the cultures, agar plates were regularly streaked with the cultures and examined under a light microscope (Olympus, model: CX40RF200) (Fig. 2). The strains were identified by the Department of Microbiology, CCSU, Meerut, and were confirmed using the keys provided by Desikachary TV (1959). The micrographs were captured using (CAMEDIA C-5060 WIDE ZOOM) and processed with ImagePro+ software (version 4.5). The physiological parameters were analyzed using a UV-Plus spectrophotometer. Purification and axenization A 100 mg of dissolved penicillin G and 50 mg of streptomycin sulphate were mixed in 10 ml of distilled water, 10 mg chloramphenicol was added in 1 ml of 95% ethanol to the penicillin-streptomycin solution. The triple antibiotic solution was filtered through a cellulose membrane (0.20 μm pore size) in a sterilized 20 ml vial and covered with aluminum foil to create axenic strains of cyanobacterial cultures (Rippka 1988). Dry weight (mg /ml) A sintered glass device was employed to homogenize and filter 50 ml of algal suspension using preweighed Whatman No. 42 filter paper. After the filtration process, the device was cooled in a desiccator and dried in an oven at 60˚C until a constant weight was achieved. The dry weight measurement was utilized to indicate the difference between the initial and final weights, serving as a key metric in the analysis (Gabrielyan et al. 2023). Estimation of pigments Using standard protocols, pigments were estimated at various stages of incubation (7 th , 14 th , 21 st , and 28 th day). Chlorophyll For estimating the chlorophyll, a homogenized algal culture of 10 ml was derived and centrifuged at 4000g for 10 minutes. The supernatant was discarded, the same amount of 95% methanol was added to the pellet, and kept in a water bath at 60 °C for 30 minutes (Mackinney 1941). The suspension was centrifuged, and the absorbance of the supernatant was measured at 650 nm and 665 nm with 95% methanol as a blank. Carotenoids For estimating the carotenoid, 10 ml of homogenized algae was centrifuged at 4000g for 10 minutes. The supernatant was discarded after rinsing with distilled water to remove any remaining salts. 2.5 ml of 85% acetone was added to the pellet and left at 4°C for one week. The suspension was subjected to freezing and thawing cycles until the supernatant turned colorless. All supernatant fractions were combined and adjusted to a final known volume (Jenssen 1978). The optical density at 450 nm was measured, using 85% acetone as a blank. Phycobiliproteins For estimating the phycobiliproteins, equivalent amounts of M K 2 HPO 4 and M KH 2 PO 4 were combined to produce a 0.05M phosphate buffer at pH 7.5, a homogeneous suspension of cyanobacterial culture of 10 ml was placed in a tube, and centrifuged at 4000 g for 10 minutes. After the removal of the supernatant, an equivalent volume of 0.05 M phosphate buffer was added to the pellet. The pigments were removed with repeated freezing and thawing (Bennett and Bogoard 1973). The optical density was measured at wavelengths of 562 nm, 615 nm, and 652 nm, using phosphate buffer as a blank. jabbrv-ltwa-all.ldf jabbrv-ltwa-en.ldf Physiochemical analysis Total soluble proteins For estimating the total soluble proteins, a standard volume of homogenized algal suspension of 1 ml was added to 5 ml of alkaline copper reagent (50 mL of 2% Na 2 CO 3 in 0.1N NaOH with 1 mL of 1% CuSO 4 .5H 2 O and 1 mL of 2% sodium potassium tartrate) and incubated at room temperature for 10 minutes at 25 °C. A 0.5 ml of Folin-Ciocalteu reagent was added and stored for 30 minutes in the dark at room temperature (Lowe and Evans 1964; Lowry et al. 1951). The absorbance of the blue colour was recorded at 660 nm using a spectrophotometer. Total carbohydrates For estimating the total carbohydrates, 1 ml of homogenized culture was added to 1 ml of 5% Anthrone solution, followed by the addition of 5 ml of concentrated sulphuric acid, and the mixture was incubated at room temperature for 10 minutes. The tubes were then placed in a water bath at 30 °C for 15 minutes (Spiro 1966). The optical density at 490 nm was recorded using a spectrophotometer. Statistical Analysis All experiments were performed in triplicate, and the results are presented as mean values ± standard deviation (SD). One-way analysis of variance (ANOVA) and Duncan’s Multiple Range Test (DMRT) were performed using IBM SPSS Statistics 25. Results Notable findings included both significant and non-significant differentiation of pigments among the different cyanobacterial strains (Table 2). Chlorophyll ranged highest of 20.0±1.0 µgmg -1 in Anabaena sp. (AS2), followed by 17.0±1.0 µgmg -1 in Nostoc sp . ( AS1), to the lowest of 2.0±1.0 µgmg -1 in Tolypothrix sp . (AS5), respectively. From Anabaena sp. (AS2) to Tolypothrix sp. (AS5), the chlorophyll levels decreased by 90%, indicating species-dependent differences in photosynthetic efficiency. Carotenoids varied from the highest of 19.0±1.0 µgmg -1 in Anabaena sp. (AS2) and the lowest of 2.0±1.0 µgmg -1 in Phormidium sp. (AS7). This indicates a 89.5% drop from Anabaena sp. to Phormidium sp., highlighting the isolate’s varying adaptive and photoprotective strategies. The different components of phycobilisomes vary significantly among all strains. PC was highest of 18±1.0 µgmg -1 in Anabaena sp. (AS2) and lowest of 2.0±1.0 µgmg -1 in Phormidium sp. (AS7), indicating that PC levels between these species had decreased by 88.9%. PE was highest in Tolypothrix sp., recording 18.0±1.0 µgmg -1 , followed by Anabaena sp. and Nostoc sp. A notable increase of more than 80% PE in Tolypothrix sp., compared to the lowest PE producer, Calothrix sp., indicates that Tolypothrix sp. has improved PE synthesis capabilities. APC was recorded highest in Tolyprothrix sp. at 20.0±1.0 µgmg -1 , followed by Phormidium sp. and Calothrix sp., ranked 2 nd and 3 rd , respectively. APC content variation indicates species-specific variations in energy transmission and light absorption. The comparative profile of carbohydrates and total soluble protein shows significant variations for all the isolates. Total soluble proteins were highest at 20±1.0 µgmg -1 in Anabaena sp. (AS2) and lowest at 2.0±1.0 µgmg -1 in Tolypothrix sp. (AS5). This equates to a 90% reduction, suggesting a significant variation in the strain’s capacities for protein production. Carbohydrates varied from the highest of 20.0±1.0 µgmg -1 in Anabaena sp. (AS2) to the lowest of 2.0±1.0 µgmg -1 in Tolypothrix sp. (AS5). The metabolic diversity was noted to be very high among strains, as it records a 90% decrease in carbohydrate as compared to the highest carbohydrate-producing strain. Anabaena sp. (AS2) exhibits the highest levels of protein, pigment synthesis, and carbohydrate storage, making it a strong contender for biotechnological applications such as carbon sequestration, pigment extraction, and biofuel production. Cyanobacterial Strains Chlorophyll Carotenoid Phycocyanin Phycoerythrin Allophycocyanin Protein Carbohydrate Anabaena sp. 20.0±1.0 a 19.0±1.0 a 18.0±1.0 a 15.0±1.0 b 11.0±1.0 d 20.0±1.0 a 20.0±1.0 a Calothrix sp. 8.0±1.0 e 8.0±1.0 e 11.0±1.0 d 2.0±1.0 f 14.0±1.0 c 8.0±1.0 e 14.0±1.0 c Nostoc sp. 17.0±1.0 b 16.0±1.0 b 13.0±1.0 c 8.0±1.0 d 8.0±1.0 e 14.0±1.0 c 11.0±1.0 d Oscillatoria sp. 11.0±1.0 d 10.0±1.0 d 16.0±1.0 b 9.0±1.0 d 5.0±1.0 f 5.0±1.0 f 8.0±1.0 e Phormidium sp. 5.0±1.0 f 2.0±1.0 g 2.0±1.0 g 5.0±1.0 e 17.0±1.0 b 11.0±1.0 d 17.0±1.0 b Tolypothrix sp. 2.0±1.0 g 5.0±1.0 f 8.0±1.0 e 18.0±1.0 a 20.0±1.0 a 2.0±1.0 g 2.0±1.0 g Westiellopsis sp. 14.0±1.0 c 13.0±1.0 c 5.0±1.0 f 12.0±1.0 c 2.0±1.0 g 17.0±1.0 b 5.0±1.0 f Table 2 Biochemical parameters (µgmg -1 ) of cyanobacterial strains with DMRT-based significance grouping (p<0.05). Fig. 1 Growth And Maintenance of Cyanobacterial Isolates AS1 Nostoc sp. AS2 Anabaena sp. AS3 Westiellopsis sp. AS4 Oscillatoria sp AS5 Tolypothrix sp. AS6 Calothrix sp. AS5 Phormidium sp Fig. 2 Photomicrographs of Strains DISCUSSION Cyanobacteria are versatile microorganisms with significant ecological and biotechnological importance, necessitating an understanding of their physiological traits (Maurya et al., 2025). This study characterized seven cyanobacterial strains, focusing on pigment analysis and physicochemical profiles. The observations were made on the 14 th day of growth in the BG-11 medium. Anabaena sp. exhibited the highest chlorophyll content (20.0±1.0 µgmg -1 ), significantly higher (p<0.05) than other strains. Chlorophyll content among strains was attributed to differences in metabolic activity, tolerance, light, and stress. Carotenoids were found at the highest concentrations in Anabaena sp. (19.0±1.0 µgmg -1 ) and the lowest in Phormidium sp (2.0±1.0 µgmg -1 ). The carotenoid content in Tolypothrix sp. is significantly larger (5.00±1.0 µgmg -1 ) than its chlorophyll content (2.00±1.0 µgmg -1 ), indicating stress-induced carotenoid accumulation, a prevalent adaptive mechanism in cyanobacteria subjected to adverse environmental conditions, including elevated light or nutritional scarcity (Yadav P et al.,2022). These pigments are valuable in biotechnology for their antioxidant properties and are useful as natural colorants in various industries. The presence of phycobiliproteins (PC, PE, and APC), which are essential for photosynthesis and have applications in food, cosmetics, and medicine (Barq et al., 2025). Anabaena sp. showed the highest amount of PC (18.0±1.0 µgmg -1 ), demonstrating its strong photosynthetic efficiency and adaptability. The findings are consistent with previous studies indicating that Anabaena sp., characterized by their filamentous and heterocystous nature, have a well-developed pigment system that enhances light absorption in diverse environmental conditions (Sharma et al., 2025). The maximum content of APC (20.0±1.0 µgmg -1 ) and PE (18.0±1.0 µgmg -1 ) was observed in Tolypothrix sp. The significant amount of PE suggests that Tolypothrix sp. could provide a competitive advantage in habitats where the quality of light changes due to variations in water depth (Kuczyńska-Kippen et al., 2024). The increased levels of APC could improve energy transfer efficiency to the photosynthetic reaction centers, aiding in consistent growth during low-light or stressful conditions (Salomon 2025). Phycobiliproteins can be highly helpful in selecting strains for prospective usage as coloring agents, phycological probes, or additives in various pharmaceutical and cosmetic products (García-Gómez et al., 2025). The protein content varied significantly among strains (p<0.05), with Anabaena sp. (20.0±1.0 µgmg -1 ) reflecting its metabolic versatility and nitrogen fixation capacity. The metabolic activity enhances its biosynthetic production, making Anabaena a strong contender as a biofertilizer, nutraceutical, and for applications involving protein in industry (Rai et al., 2025). As the incubation time increased, the amount of carbohydrates also increased. Anabaena sp. had the highest carbohydrate content at (20.0±1.0 µgmg -1 ), while Tolypothrix sp. had the lowest (2.0±1.0 µgmg -1 ). It is clear that the average daily growth and biomass directly impact the carbohydrate content. They serve as substrates for carbon fixation and the synthesis of organic molecules, playing a role in the process of photosynthesis (Karishma et al., 2024). The importance of this comparative analysis is rooted in its capacity to pinpoint metabolically advanced strains for specific applications in biofertilizers, natural colorants, protein supplements, and renewable energy sources. It addresses a significant gap in comparative physiological profiling, offering a valuable reference for forthcoming selection and optimization endeavors in sustainable biotechnology. Conclusion The cyanobacterial strains, based on their pigment composition and metabolic characteristics, are being explored for potential biotechnology applications. Anabaena sp. is a promising candidate for biofuel generation, natural colorants, and pharmaceuticals. Artificial colorants pose environmental and health risks, so natural sources like plants or algae are a desirable substitute. Tolypothrix sp. could be used in food, cosmetics, and fluorescence-based biosensors. Nostoc sp., Westiellopsis sp., and Oscillatoria sp. could be used for environmental remediation and biofertilizer manufacturing. Further research should focus on optimizing growth conditions, genetic changes, and large-scale farming techniques. The comparative physiological assessment done in this research increases our scientific understanding of cyanobacterial diversity and functionality and bears practical implications for boosting the sustainability and efficiency of agricultural and industrial systems. Acknowledgments I am thankful to SRM Institute of Science and Technology, Delhi-NCR Campus, Modinagar, Ghaziabad, 201204, U.P., India, and Chaudhary Charan Singh University, Meerut, 250001, U.P., India, for providing me with the research platform and availability, which was helpful during my research work. Funding No authors have specific funding for this work. 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Keywords cyanobacterial strains phycobiliproteins physiochemical analysis physiological parameters pigment composition Authors Affiliations Anjali Singh 0009-0001-1473-7412 SRM Institute of Science and Technology - Delhi NCR Campus View all articles by this author Kamakshi [email protected] SRM Institute of Science and Technology - Delhi NCR Campus View all articles by this author Devendra Kumar Chaudhary Charan Singh University View all articles by this author Metrics & Citations Metrics Article Usage 244 views 139 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Anjali Singh, Kamakshi, Devendra Kumar. The Comparison of Physiological Parameters of seven different Cyanobacterial strains. Authorea . 25 July 2025. DOI: https://doi.org/10.22541/au.175345758.85764042/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. 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