Applications of Sentinel-2 satellite data for spatio-temporal mapping of deep pools for monitoring the riverine connectivity and assessment of ecological dynamics: a case from Godavari, a tropical river in India (2016-2021).

Rivers are dynamic ecosystems with diverse habitats that require ample connectivity to ensure the flow of ecosystem services, thus empowering the sustainable development of an entire basin. Geo-spatial tools offer powerful prospects for monitoring of aquatic ecosystems. The usefulness of Sentinel-2 datasets to assess river connectivity has been explored for an un-gauged seasonal river system. The present study was undertaken in five ecologically unique river reaches viz. Wainganga, Wardha, Pranhita, Godavari-mid and Manair in Godavari Basin in the Indian Deccan Plateau to map water spread dynamics at various time scales, i.e., fortnightly, monthly, seasonal, annual and demi-decadal during 2016-2021. The maximum value of perennial water spread per square kilometre of total floodplain area (2016-2021), determined using Sentinel-2 imageries, was observed in river Wardha (0.18) followed by Pranhita (0.12) and Wainganga (0.11). The water spread showed a decreasing trend, while the number of patches in the river corridor increased over time from post-monsoon to pre-monsoon season. The copious perennial habitat with relatively larger patches, incessant flow in river Pranhita and obstructed flow, large-sized patches reported in river Wardha during summer months, hold importance in terms of providing refuge to aquatic biota. This study provides evidence for the impact of water projects on spatio-temporal water spread dynamics in Godavari Basin. The demonstrated utility of Sentinel-2 imagery coupled with gauge station measurements for river continuity assessment and deep pool mapping would aid in enhancing our understanding on environmental flow at a spatial scale, which in turn would aid in effective river management to achieve the Sustainable Development Goals. The implications of this study for sustainable environmental management and limitations are also discussed.

Triclosan induces immunosuppression and reduces survivability of striped catfish Pangasianodon hypophthalmus during the challenge to a fish pathogenic bacterium Edwardsiella tarda.

Toxicological studies on the emergent pollutant, triclosan (TCS) have established the wide-ranging effects of the compound on fish and other aquatic organisms. Although the available literature describes the standalone effects of TCS on growth and metabolism of fish yet, reports about the combined effects of TCS with microbial pathogens are scarce. In a real environment, a combined exposure to TCS and pathogens is of common occurrence, therefore, such investigation facilitates in developing a better understanding about the gross effects of pollutants and microbial pathogens on aquatic organisms including fish. In this context, the experimental fish (striped catfish, Pangasianodon hypophthalmus) were exposed to three different concentrations of TCS viz. 10, 20 and 30% of 96 h LC50 (1177 µg L-1) for 45 days including two control group firstly solvent control (without TCS) group and another one (without solvent and TCS) group in triplicate. Sampling was performed fortnightly and blood, serum and tissues (liver, and gills) samples were collected for evaluating immunological and biochemical parameters. Following 45 days of the experiments, the experimental fish in each treatment group including controls were challenged with a fish pathogenic bacterium Edwardsiella tarda (LD50 dose) and fish mortality was daily monitored for calculating cumulative mortality till 7 days and further, relative per cent survivable was estimated. A significant reduction in cellular immune responses i.e. respiratory burst activity (RBA), myeloperoxidase activity (MPO), phagocytic activity (PA) and humoral immune components viz. serum lysozyme activity, total immunoglobulin in serum, ceruloplasmin level, serum total protein, albumin and globulin level was evident in TCS exposed groups in comparison to control during the experimental periods. Further, oxidative stress parameters viz. superoxide dismutase (SOD), catalase (CAT), glutathione-s-transferase (GST) activity in liver and gill tissue exhibited a dose-dependent increase in activity with related to TCS concentration during the experimental periods. A significant reduction in relative percentage survival was observed with increasing TCS concentration. The present study reveals that TCS can inhibit the cellular and humoral components of the innate immune system of the fish and can elevate the mortality due to TCS mediated immunosuppression in fish during the bacterial infection.

Triclosan exposure induces varying extent of reversible antimicrobial resistance in Aeromonas hydrophila and Edwardsiella tarda.

Triclosan (TCS) is a biocide commonly used in household and personal care items to prevent the microbial growth and is currently considered as an emerging pollutant. It has a ubiquitous distribution which can substantially contribute towards antimicrobial resistance. The present study was designed to evaluate the effect of different concentrations of TCS exposure on the antibiotic sensitivity of aquatic bacteria. Aeromonas hydrophila ATCC® 49140™ and Edwardsiella tarda ATCC® 15947™ exposed to TCS for short (30 min) and long duration (serial passages). The agar-disc diffusion assay during the serial passages of TCS exposure and subsequent exposure withdrawal showed clinically insignificant changes in the zone of inhibition for six selected antibiotics in both bacterial strains at all exposure concentrations. Four folds concentration-dependent increase in the minimum inhibitory concentrations (MICs) of TCS was observed in both the strains following TCS exposure. Similarly, a concentration-dependent increase in the MICs of oxytetracycline (OTC) up to 4 folds in A. hydrophila, and up to 8 folds in E. tarda, was also documented during the TCS exposure. In all the cases, withdrawal of TCS exposure effectively reduced the MICs of TCS and OTC in blank passages suggesting a decline in acquired resistance. The frequencies of mutation during 30 min TCS exposure for E. tarda and A. hydrophila ranged between >10-6 and 10-7 levels. Nevertheless, the TCS exposure did not cause any detectable mutation on the fabV gene of A. hydrophila indicating that the TCS may elicit phenotypic adaptation or other resistance mechanism. Although the reduction in MICs due to exposure withdrawal did not restore the bacterial susceptibility up to the initial level, the study proved that the reduced TCS use could significantly help reduce the antimicrobial-resistance and cross-resistance in pathogenic bacteria.

Physical entrapment of chitosan in fixed-down-flow column bed enhances triclosan removal from water.

Triclosan (TCS) is an emergent pollutant with wide-ranging deleterious effects on aquatic organisms and humans. There is a growing concern about the development of low-cost and efficient treatment systems for the removal of TCS from water. This report describes the performance of a prototype of a continuous flow, fixed bed column device with physically entrapped industrial by-product chitosan. The effects of initial TCS concentration, adsorbent dose in the column matrix, and flow rate were investigated with regard to removal efficiency (%), adsorption capacity and breakthrough time. To understand the thermodynamic properties of the adsorption process, three kinetic models - Thomas, Yoon-Nelson and Adams-Bohart - were applied to the experimental data for the prediction of characteristic parameters of the adsorption process. The Yoon-Nelson model showed the best agreement between the experimental and calculated values. The column showed a near saturation state (Ct/C0 = 0.92; C0 and Ct are the concentration of TCS before and after treatment.) at 90 mg L-1 TCS concentration after 60 minutes. In view of non-availability of a treatment process for the emergent pollutant TCS, the data of the present investigation will facilitate the development of novel prototypes of column bed reactors for the removal of TCS.

Toxic Effects of Selected Textile Dyes on Elemental Composition, Photosynthetic Pigments, Protein Content and Growth of a Freshwater Chlorophycean Alga Chlorella vulgaris.

Toxicity of three textile dyes-Optilan yellow, Drimarene blue and Lanasyn brown, was evaluated in a green alga Chlorella vulgaris. The unialgal populations of the alga showed a concentration-dependent decrease in specific growth rate and pigments after exposure to graded concentrations of above dyes. The elemental profile (C, H, N, S) of the treated and untreated cells showed a change which was evident from a significant decrease in the quantity of elements after exposure to the dyes. The observations provide convincing evidence that the textile dyes inhibited the growth, pigment and elemental composition of the algal cells. The findings of the present investigation will contribute to gaining a better understanding of the impacts of textile dyes on ecologically important aquatic organisms.

A prototype of novel agro-waste based column bed device for removal of textile dye Optilan Red.

The aim of the study was to assess the potentiality of an agro-waste (sugarcane bagasse) for removal of the textile dye (Optilan Red) using novel column based filtration unit with a packed column of chemically treated sugarcane bagasse. The treated and untreated sugarcane bagasse (biosorbent) were characterized by Fourier transform infrared (FT-IR) spectroscopy. Effect of initial dye concentration on percentage removal of dye, equilibrium adsorption of sugarcane bagasse, kinetic studies, breakthrough point equilibrium and desorption of dye from the column material were studied. An inverse dependence of initial dye concentration on percent removal of dye was observed, whereas the equilibrium adsorption (qe) showed a direct relationship with dye concentration. The time required for reaching breakthrough point was 120 min. Desorption of dye through alkali wash resulted in complete desorption after 1 h washing of the column for its reuse for next cycle. FT-IR analysis shows vibration in valence bands of the hydrogen bond of OH group, and the bands of intra-molecular and intermolecular hydrogen bonds, which results in interaction of treated bagasse with Optilan Red textile dye. The present study showed that more than 93% removal of the dye can be achieved in the concentration range 10-50 ppm (aqueous solution). The removal efficiency of the column remained almost unchanged for the treatment of dye-house wastewater spiked with the dye. The agro-waste based treatment process shows a considerable potential for a low-cost treatment of dye contaminated water.

Biomarkers-based assessment of triclosan toxicity in aquatic environment: A mechanistic review.

Triclosan (TCS), an emergent pollutant, is raising a global concern due to its toxic effects on organisms and aquatic ecosystems. The non-availability of proven treatment technologies for TCS remediation is the central issue stressing thorough research on understanding the underlying mechanisms of toxicity and assessing vital biomarkers in the aquatic organism for practical monitoring purposes. Given the unprecedented circumstances during COVID 19 pandemic, a several-fold higher discharge of TCS in the aquatic ecosystems cannot be considered a remote possibility. Therefore, identifying potential biomarkers for assessing chronic effects of TCS are prerequisites for addressing the issues related to its ecological impact and its monitoring in the future. It is the first holistic review on highlighting the biomarkers of TCS toxicity based on a comprehensive review of available literature about the biomarkers related to cytotoxicity, genotoxicity, hematological, alterations of gene expression, and metabolic profiling. This review establishes that biomarkers at the subcellular level such as oxidative stress, lipid peroxidation, neurotoxicity, and metabolic enzymes can be used to evaluate the cytotoxic effect of TCS in future investigations. Micronuclei frequency and % DNA damage proved to be reliable biomarkers for genotoxic effects of TCS in fishes and other aquatic organisms. Alteration of gene expression and metabolic profiling in different organs provides a better insight into mechanisms underlying the biocide's toxicity. In the concluding part of the review, the present status of knowledge about mechanisms of antimicrobial resistance of TCS and its relevance in understanding the toxicity is also discussed referring to the relevant reports on microorganisms.

A multi-biomarker approach using integrated biomarker response to assess the effect of pH on triclosan toxicity in Pangasianodon hypophthalmus (Sauvage, 1878).

Application of biomarkers is an effective approach for a better understanding of varying toxicity in aquatic organisms during the seasonal and diurnal changes in the natural environment. This report describes the toxicity of sub-lethal concentrations of triclosan (TCS) at different pH (6.5, 7.5 and 8.5) based on selected biomarkers related to oxidative stress, metabolism and genotoxicity in Pangasianodon hypophthalmus. The 96 h LC50 of TCS for P. hypophthalmus was lower at pH 6.5 when compared to higher pH. The sub-lethal concentration of TCS exhibited a significant decrease in hematological parameters related to complete blood counts except for total leukocyte count (TLC), mean cell haemoglobin concentration (MCHC) and red cell distribution width (RDW). Multivariate data analysis showed a significant interaction of TCS and pH in metabolizing enzymes like glutamic oxaloacetate transaminase (GOT), glutamic pyruvic transaminase (GPT), Lactate dehydrogenase (LDH), antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione-s-transferase (GST) and neurotransmitter enzyme acetylcholinesterase (AChE). A significant increase in DNA damage and micronuclei frequency in liver and blood cells of TCS exposed fish at pH 6.5 indicate that the TCS exposure has pronounced effects on genetic materials. The findings of present study establish that enzymes like SOD, LDH, GOT, AChE, DNA damage and micronuclei frequency can be successfully deployed as biomarkers for the assessment of toxicity of TCS in fish.

Effect of temperature on triclosan toxicity in Pangasianodon hypophthalmus (Sauvage, 1878): Hematology, biochemistry and genotoxicity evaluation.

The rising level of triclosan (TCS) in aquatic environment is raising concerns and in this context, evaluation of toxicity towards aquatic organisms under varying environmental conditions, especially temperature, is a pre-requisite for a better understanding of the toxic effects on specific metabolic processes. In this report, the mechanistic physiological responses of fish towards varying concentration of TCS at graded temperature were evaluated. The static renewal acute test was performed, and 96 h median lethal concentration (LC50) of TCS for Pangasianodon hypophthalmus was estimated and the values were 848.33, 1181.94 and 1356.96 µg L-1 at 25, 30 and 35 °C respectively. The chronic study was performed for 30 days at 1/5th and 1/10th concentration of the estimated LC50 of TCS at 25, 30 and 35 °C respectively. The chronic effects resulted in significant decrease in total erythrocyte count (TEC), hemoglobin (Hb), packed cell volume (PCV), mean corpuscular hemoglobin (MCH) and mean cell volume (MCV), while a significant increase in total leukocyte count (TLC), mean corpuscular hemoglobin concentration (MCHC) and red cell distribution width (RDW) was observed in TCS exposed groups at 25-35 °C. Further, a significant increase in activity of transaminase enzymes, lactate dehydrogenase (LDH) and antioxidant enzymes (superoxide dismutase) (SOD) and catalase (CAT) except glutathione-S-transferase (GST) in liver and acetylcholinesterase (AChE) in brain of the TCS exposed fish was recorded in all the above temperature range. Severe damage of DNA in nucleus of blood and liver cells, and high micronuclei frequency (MNi) was noticed in TCS exposed groups at 25 °C. The report provides convincing evidence for the effect of temperature on TCS toxicity. The findings will help in gaining a better insight into the change in toxicity of TCS in a natural environment where diurnal variations in temperature may be crucial in determining the overall extent of toxicity.

Triclosan toxicity alters behavioral and hematological parameters and vital antioxidant and neurological enzymes in Pangasianodon hypophthalmus (Sauvage, 1878).

Triclosan and its metabolites are detected in a diverse aquatic environment and are major concerns for various aquatic organisms. The present study investigated the impact of acute and sub-lethal exposure of triclosan on behaviour, activities of acetylcholinesterase and selected antioxidant enzymes, haematological and serum gas-electrolyte parameters of Pangasianodon hypophthalmus. The 96 h LC50 of triclosan for P. hypophthalmus was estimated as 1458 µg L-1. Further, sub-lethal triclosan exposure to 1/15th (97 µg L-1), 1/10th (145 µg L-1) and 1/5th (291 µg L-1) of 96 h LC50 concentration for a period of 45 days lead to decrease in total erythrocyte count, haemoglobin content and packed cell volume of blood while total leukocyte count increased significantly (p < 0.05) as compared to control. A concentration-dependent increase in the serum chloride and decrease in partial pressure of oxygen in blood serum was noted on 45th day. An increased activity of catalase and superoxide dismutase in gill and liver tissues and inhibition of acetylcholinesterase activity in brain was observed on 15th, 30th and 45th day of exposure which was dependent on both - concentration of triclosan and duration of exposure. A significant high activity of glutathione-S-transferase in gill and liver tissue was observed in triclosan exposed groups in comparison to control during the experimental period. The study shows that long-term sub-lethal exposure of triclosan to fish can lead to several physiological alterations such as enzymatic scavenging of oxygen radicals and the normal neurological functions mediated by the enzyme acetylcholinesterase. With increasing anthropogenic activity, the study provides a convincing evidence for the necessity of a regulated use and safer disposal of triclosan to the environment.

Soil quality index for evaluation of reclaimed coal mine spoil.

Success in the remediation of mine spoil depends largely on the selection of appropriate tree species. The impacts of remediation on mine soil quality cannot be sufficiently assessed by individual soil properties. However, combination of soil properties into an integrated soil quality index provides a more holistic status of reclamation potentials of tree species. Remediation potentials of four tree species (Acacia auriculiformis, Cassia siamea, Dalbergia sissoo, and Leucaena leucocephala) were studied on reclaimed coal mine overburden dumps of Jharia coalfield, Dhanbad, India. Soil samples were collected under the canopies of the tree species. Comparative studies on the properties of soils in the reclaimed and the reference sites showed improvements in soil quality parameters of the reclaimed site: coarse fraction (-20.4%), bulk density (-12.8%), water holding capacity (+0.92%), pH (+25.4%), EC (+2.9%), cation exchange capacity (+46.6%), organic carbon (+91.5%), N (+60.6%), P (+113%), K (+19.9%), Ca (+49.6%), Mg (+12.2%), Na (+19.6%), S (+46.7%), total polycyclic aromatic hydrocarbons (-71.4%), dehydrogenase activity (+197%), and microbial biomass carbon (+115%). Principal component analysis (PCA) was used to identify key mine soil quality indicators to develop a soil quality index (SQI). Selected indicators include: coarse fraction, pH, EC, soil organic carbon, P, Ca, S, and dehydrogenase activity. The indicator values were converted into a unitless score (0-1.00) and integrated into SQI. The calculated SQI was significantly (P<0.001) correlated with tree biomass and canopy cover. Reclaimed site has 52-93% higher SQI compared to the reference site. Higher SQI values were obtained for sites reclaimed with D.sissoo (+93.1%) and C.siamea (+86.4%).

Assessment of the effect of the toxicity of a textile dye on Nostoc muscorum ISU, a diazotrophic cyanobacterium.

The impact of graded concentrations (5, 10, 15 and 20 mg dm(-3) of acid mordant Metomega Chrome Orange GL, a common dye used in woollen, carpet and textile industries, was studied on protein and pigment content and photosynthetic oxygen evolution in a diazotrophic cyanobacterium Nostoc muscorum. The lower concentration of 5 mg dm(-3) showed a negligible effect on protein and pigment content and photosynthetic oxygen evolution, whereas at higher concentrations a drastic decrease in the above parameters was observed. The highest concentration of 20 mg dm(-3) resulted in a decrease of protein, chlorophyll a, phycocyanin and carotenoid content by 72, 76, 54 and 17% of the control, respectively. Photosynthetic oxygen evolution also decreased by 92% of the control at this concentration of the dye.

Impact of endosulfan on cytoplasmic and mitochondrial liver malate dehydrogenase from the freshwater catfish (Clarias batrachus).

The impact of a sublethal concentration of an organochlorine pesticide endosulfan on the activity, specific activity, electrophoretic patterns and kinetic properties of crude and purified forms of cytoplasmic malate dehydrogenase (cMDH) and mitochondrial malate dehydrogenase (mMDH) was evaluated in the liver of the freshwater catfish, Clarias batrachus. The endosulfan reduced significantly the activity and the specific activity of cMDH and mMDH, but had no effect on total cytoplasmic and mitochondrial protein contents. The inhibition produced by endosulfan was of mixed non-competitive-uncompetitive type (KiE > KiES) and of mixed competitive-non-competitive type (KiE < KiES) for crude cMDH and mMDH, respectively. The PAGE shows five distinct isoforms (C1 to C5) of cMDH and two isoforms (M1 and M2) of mMDH. The C5-isoform of liver cMDH is predominant and it corresponds to M2-isoform of mMDH. There are no endosulfan-associated differences in the relative charges of crude cMDH and mMDH as well as their purified isoforms, C5-cMDH and M2-mMDH. The relative molecular weights of the purified isoforms are not affected by endosulfan. The purified C5-cMDH and M2-mMDH of endosulfan-treated liver in vivo showed simple non-competitive (KiE = KiES) type of inhibition; whereas in vitro it was of uncompetitive (KiES) and mixed competitive-non-competitive (Ki < KiES) type for the two respective isoforms. G-1-P acts as an uncompetitive (KiES) inhibitor of C5-cMDH and mixed competitive-non-competitive (KiE < KiES) inhibitor of M2-mMDH of the control fish. The inhibitory pattern of G-1-P is modulated by endosulfan in case of C5-cMDH; whereas there is no alteration in case of M2-mMDH. Summarizing, it can be stated that endosulfan exerts an inhibitory effect on crude cMDH and mMDH in vivo, and their purified isoforms (C5-cMDH and M2-mMDH) in vivo as well as in vitro. The impact of endosulfan is mediated through enzyme-substrate-endosulfan (ES-END) complexing for cMDH and enzyme-endosulfan (E-END) complexing for mMDH.

Prenatal mortality and its effects on subsequent fertility in purebred and crossbred Gir cows.

A study was conducted on 851 cows (Gir cows and their crossbreeds) between 1971 and 1984 at the Livestock Farm of the All India Coordinated Research Project on Cattle in Jabalpur, India. The cows were impregnated with frozen or fresh semen from 90 bulls and a total of 2699 pregnancies resulted. An overall incidence of prenatal mortality was recorded at 4.89%. A significantly higher (P / 0.01) incidence of prenatal mortality was encountered in three breed crosses in comparison with Gir cows and their first generation crossbreeds. Parity of dam had no effect on prenatal mortality. However, effects of season and individual sires on prenatal mortality were documented. It was observed that reproductive performance was improved significantly after prenatal mortality.

Malate and lactate dehydrogenases of a freshwater catfish: impact of endosulfan.

A sublethal concentration of technical grade endosulfan (END) inhibited 35 to 55% of the activities of cytoplasmic malate dehydrogenase (cMDH), mitochondrial malate dehydrogenase (mMDH), and lactate dehydrogenase (LDH) in the liver and the skeletal muscle of a freshwater catfish, Clarias batrachus, after 7 days of exposure. The activity remained in the inhibited state up to 28 days. The withdrawal of END from the medium after 1 week of exposure gradually restored the activities to control levels within 21 days in the skeletal muscle and 28 days in the liver. The administration of actinomycin D or cycloheximide between the 14th and the 21st day of the withdrawal of END almost completely inhibited the withdrawal-dependent recovery in the activities of all the three enzymes. This indicates de novo synthesis of the enzymes during the recovery period. A conjoint treatment of END and triiodothyronine (T3) raised the activities of cMDH, mMDH, and LDH in the liver and the skeletal muscle up to the control levels. This shows that the inhibitory effect of END may be relieved in presence of T3. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed few changes in the pattern of cytoplasmic proteins of the liver and the skeletal muscle in response to exposure to END.

Enzymatic and ultrastructural studies in a freshwater catfish: impact of methyl parathion.

Exposure to a sublethal concentration of methyl parathion (MEP) reduced the activity of cytoplasmic malate dehydrogenase, mitochondrial malate dehydrogenase, and lactate dehydrogenase by 30 to 49% in the liver and the skeletal muscle of the freshwater catfish. Clarias batrachus, after 7 days. The activities then began to recover and reached the control levels on the 28th day of MEP exposure. A complete recovery occurred on the 7th day when MEP was withdrawn from the medium after an exposure for 1 week. The withdrawal-dependent recovery in the activities was inhibited partially or completely by actinomycin D and cycloheximide, suggesting de novo synthesis of the enzymes during the recovery period. A conjoint treatment of MEP and triiodothyronine (T3) restored the activities to control levels, indicating T3 protection against the pesticide toxicity. SDS-PAGE of the cytoplasmic fraction of the liver showed some noticeable changes in the protein pattern after an exposure to MEP. Ultrastructural studies on MEP-treated liver cells showed disappearance of the glycogen granules and appearance of numerous smooth endoplasmic reticulum, lysosomal dense bodies, and swollen mitochondria. These changes in the liver are an indication of hepatic toxicity leading toward necrosis.

An assessment of biopotential of three cyanobacterial isolates from Antarctic for carotenoid production.

Specific growth rates and carotenoid contents of three Antarctic and tropical strains of cyanobacteria viz. Anabaena sp., Phormidium sp. and Nostoc sp. were compared in batch and mass cultures to assess bio-potential of Antarctic strains for cost-effective carotenoid production. Antarctic strains though exhibited slightly lower specific growth rates, but contained higher carotenoid contents (per unit dry wt.), than tropical strains. Modification of normal composition of BG-11 culture medium, by altering nitrogen and carbon sources resulted in 25-38% increase in carotenoid content in both types of strains. Mass-culture in indoor and semi-outdoor bio-reactors resulted in 39-113% higher carotenoid content in Antarctic strains, compared to their respective tropical strains. The observations suggest that Antarctic cyanobacteria may have potential as superior strains for maximizing the yield of carotenoids.

Chlorella mirabilis as a Potential Species for Biomass Production in Low-Temperature Environment.

Successful adaptation/acclimatization to low temperatures in micro-algae is usually connected with production of specific biotechnologically important compounds. In this study, we evaluated the growth characteristics in a micro-scale mass cultivation of the Antarctic soil green alga Chlorella mirabilis under different nitrogen and carbon sources followed by analyses of fatty acid contents. The micro-scale mass cultivation was performed in stable (in-door) and variable (out-door) conditions during winter and/or early spring in the Czech Republic. In the in-door cultivation, the treatments for nitrogen and carbon sources determination included pure Z medium (control, Z), Z medium + 5% glycerol (ZG), Z medium + 5% glycerol + 50 µM KNO3 (ZGN), Z medium + 5% glycerol + 200 µM NH4Cl (ZGA), Z medium + 5% glycerol + 1 mM Na2CO3 (ZNC), Z medium + 5% glycerol + 1 mM Na2CO3 + 200 µM NH4Cl (ZGCA) and Z medium + 5% glycerol + 1 mM Na2CO3 + 50 µM KNO3 (ZGCN) and were performed at 15°C with an irradiance of 75 µmol m(-2) s(-1). During the out-door experiments, the night-day temperature ranged from -6.6 to 17.5°C (daily average 3.1 ± 5.3°C) and irradiance ranged from 0 to 2,300 µmol m(-2) s(-1) (daily average 1,500 ± 1,090 µmol m(-2) s(-1)). Only the Z, ZG, ZGN, and ZGC treatments were used in the out-door cultivation. In the in-door mass cultivation, all nitrogen and carbon sources additions increased the growth rate with the exception of ZGA. When individual sources were considered, only the effect of 5% glycerol addition was significant. On the other hand, the growth rate decreased in the ZG and ZGN treatments in the out-door experiment, probably due to carbon limitation. Fatty acid composition showed increased production of linoleic acid in the glycerol treatments. The studied strain of C. mirabilis is proposed to be a promising source of linoleic acid in low-temperature-mass cultivation biotechnology. This strain is a perspective model organism for biotechnology in low-temperature conditions.