Search results for: cyanobacteria

Authors: Hugo F. Lobaton

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The cyanobacterium Spirulina platensis is a prokaryotic photoautotrophic microorganism that is successfully cultivated for the commercialization as whole biomass due to its high protein content and promising valuable substance. For instance, phycocyanin has recently drawn the interest of the food and cosmetic industries due to its bright blue colour and its strong antioxidant capacities. The phycocyanin (PC) is the main protein-pigment in S. platensis (4% to 20%). In batches, the rate of overproduction of metabolites by cyanobacteria is limited or activated by the depletion of required substrates. The aim of this study was to develop a kinetic law that describes phycocyanin formation during batch cultivation. S. platensis was cultivated in 1 L bubble column photobioreactor with 30°C and 700 µmol m⁻² s⁻¹. Culture samples were daily collected from the bubble columns in sterile conditions. The biomass (g l⁻¹) was measured directly after a biomass lyophilisation process, and phycocyanin extractions and measurements were done according to a well-established protocol. A kinetic law for phycocyanin formation that includes nitrate and bicarbonate limitations was proposed and linked to the biomass core model. The set of differential equations were solved in MATLAB. Concerning to product formation, the experimental results show that phycocyanin mass fraction is degraded as results of the complete nitrate depletion and nitrate additions during the cultivation help to keep constant this molecule until new macro-element limitation appear. According to the model, bicarbonate is this limitation.

Keywords: phycocyanin, nitrate, bicarbonate, spirulina

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Authors: Anurakti Shukla, Sudhakar Srivastava

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Cyanobacteria are known for rapid growth rates, high biomass, and the ability to accumulate potentially toxic elements and contaminants. The present work was planned to develop a low-cost, feasible prototype, CyanoClean, for the growth of a cyanobacterial consortium for the removal of arsenic (As) from water. The cyanobacterial consortium consisting of Oscillatoria, Phormidiumand Gloeotrichiawas used, and the conditions for optimal growth of the consortium were standardized. A pH of 7.6, initial cyanobacterial biomass of 10 g/L, and arsenite [As(III)] and arsenate [As(V)] concentration of 400 μΜand 600 μM, respectively, were found to be suitable. The CyanoClean prototype was designed with acrylic sheet and had arrangements for optimal cyanobacterial growth in natural sunlight and also in artificial light. The As removal experiments in concentration- and duration-dependent manner demonstrated removal of up to 39-69% and 9-33% As respectively from As(III) and As(V)-contaminated water. In field testing of CyanoClean, natural As-contaminated groundwater was used, and As reduction was monitored when a flow rate of 3 L/h was maintained. In a field experiment, As concentration in groundwater was found to reduce from 102.43 μg L⁻¹ to <10 μg L⁻¹ after 6 h in natural sunlight. However, in shaded conditions under artificial light, the same result was achieved after 9 h. The CyanoClean prototype is of simple design and can be easily up-scaled for application at a small- to medium-size land and shall be affordable even for a low- to middle-income group farmer.

Keywords: cyanoclean, gloeotrichia, oscillatoria, phormidium, phycoremediation

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Authors: Jou-An Chen, Julio Castillo, Errol Duncan Cason, Gabre Kemp, Leana Esterhuizen, Angel Valverde Portal, Esta Van Heerden

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Emerging contaminants (EC) such as pharmaceutical and personal care products have been accumulating for years in water bodies all over the world. However, very little is known about the occurrences, levels, and effects of ECs in South African water resources. This study provides an initial assessment of the distribution of eight ECs (Acetaminophen, Atrazine, Terbuthlyazine, Carbamazepine, Phenyton, Sulfmethoxazole, Nevirapine and Fluconozole) in fifteen water sources from the Free State and Easter Cape provinces of South Africa. Overall, the physiochemical conditions were different in surface and groundwater samples, with concentrations of several elements such as B, Ca, Mg, Na, NO3, and TDS been statistically higher in groundwater. In contrast, ECs levels, quantified at ng/mL using the LC/MS/ESI, were much lower in groundwater samples. The ECs with higher contamination levels were Carbamazepine, Sulfmethoxazole, Nevirapine, and Terbuthlyazine, while the most widespread were Sulfmethoxazole and Fluconozole, detected in all surface and groundwater samples. Fecal and E. coli tests indicated that surface water was more contaminated than groundwater. Microbial communities, assessed using NGS, were dominated by the phyla Proteobacteria and Bacteroidetes, in both surface and groundwater. Actinobacteria, Planctomycetes, and Cyanobacteria, were more dominant in surface water, while Verrucomicrobia were overrepresented in groundwater. In conclusion, ECs contamination is closely associated with human activities (human wastes). The microbial diversity identified can suggest possible biodegradation processes.

Keywords: emerging contaminants, EC, personal care products, pharmaceuticals, natural attenuation process

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Authors: Ravi Raghav Sonani, Niraj Kumar Singh, Jitendra Kumar, Datta Madamwar

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Phycobiliproteins (PBPs) are light harvesting proteins showing very strong absorbance and fluorescence in the visible range of the solar spectrum. Phycoerythrin (PE) and phycocyanin (PC) are majorly found PBPs in the cyanobacteria and red algae. In the present study, we have investigated the reactive oxygen species (ROS)-averting capacity of purified PE and PC of cyanobacterial origin. Furthermore, the possibility - whether the ROS-averting potential of PBPs can be explored in the therapeutics of oxidative stress associated physiological anomalies including aging and neurodegenerative diseases. The nematode Caenorhabditis elegans has been used as model organism in this study. PE and PC treatment moderated normal aging and associated physiological functionalities like pharyngeal pumping and locomotion of C. elegans. Moreover, PE-treatment enhanced the stress (oxidative and heat) tolerance upon PE and PC treatment. Specifically, PE treatment was also noted to moderate the progression of Alzheimer’s disease in transgenic C. elegans CL4176. However, PC-treatment curtailed the polyQ aggregation mediated proteotoxicity in C. elegans AM141 (Huntington disease model) under stressed (paraquat stress) as well as normal conditions. The effectiveness of PE and PC in expanding the lifespan of mutant C. elegans knockout for some up- (daf 16) and down- (daf-2 and age-1) stream regulators of insulin/IGF-1 signalling (IIS) shows the independency of their effects from DAF-2–AGE-1–DAF-16 signalling pathway. In conclusion, the present report demonstrates the anti-aging and neuro-protective potential of cyanobacterial PE and PC.

Keywords: phycobiliproteins, aging, alzheimer, huntington, C. elegans

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Authors: Raymond Ezenweani, Jeffrey Ogbebor

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The menace of aquatic pollution has become increasingly of great concern and the effects of this pollution as a result of anthropogenic activities cannot be over emphasized. Phycoremediation is the application of algal remediation technology in the removal of harmful products from the environment. Harmful products also known as pollutants are usually introduced into the environment through variety of processes such as industrial discharge, agricultural runoff, flooding, and acid rain. This work has to do with the capability of algae in the efficient removal of different pollutants, ranging from hydrocarbons, eutrophication, agricultural chemicals and wastes, heavy metals, foul smell from septic tanks or dumps through different processes such as bioconversion, biosorption, bioabsorption and biodecomposition. Algae are capable of bioconversion of environmentally persistent compounds to degradable compounds and also capable of putting harmful bacteria growth into check in waste water remediation. Numerous algal organisms such as Nannochloropsis spp, Chlorella spp, Tetraselmis spp, Shpaerocystics spp, cyanobacteria and different macroalgae have been tested by different researchers in laboratory scale and shown to have 100% efficiency in environmental remediation. Algae as a result of their photosynthetic capacity are also efficient in air cleansing and management of global warming by sequestering carbon iv oxide in air and converting it into organic carbon, thereby making food available for the other organisms in the higher trophic level of the aquatic food chain. Algae play major role in the sustenance of the aquatic ecosystem by their virtue of being photosynthetic. They are the primary producers and their role in environmental sustainability is remarkable.

Keywords: Algae , Pollutant, ., Phycoremediation, Aquatic, Sustainability

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Authors: Sean T. S. Wei, Joy D. Van Nostrand, Annapoorna Maitrayee Ganeshram, Stephen B. Pointing

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McMurdo Dry Valleys are a largely ice-free polar desert protected by international treaty as an Antarctic special managed area. The terrestrial landscape is dominated by oligotrophic mineral soil with extensive rocky outcrops. Several environmental stresses: low temperature, lack of liquid water, UV exposure and oligotrophic substrates, restrict the major biotic component to microorganisms. The bacterial diversity and the putative physiological capacity of microbial communities of quartz rocks (hypoliths) and soil of a maritime-influenced Dry Valleys were interrogated by two metagenomic approaches: 454 pyro-sequencing and Geochp DNA microarray. The most abundant phylum in hypoliths was Cyanobacteria (46%), whereas in solils Actinobacteria (31%) were most abundant. The Proteobacteria and Bacteriodetes were the only other phyla to comprise >10% of both communities. Carbon fixation was indicated by photoautotrophic and chemoautotrophic pathways for both hypolith and soil communities. The fungi accounted for polymer carbon transformations, particularly for aromatic compounds. The complete nitrogen cycling was observed in both communities. The fungi in particular displayed pathways related to ammonification. Environmental stress response pathways were common among bacteria, whereas the nutrient stress response pathways were more widely present in bacteria, archaea and fungi. The diversity of bacterialphage was also surveyed by Geochip. Data suggested that different substrates supported different viral families: Leviviridae, Myoviridae, Podoviridae and Siphoviridiae were ubiquitous. However, Corticoviridae and Microviridae only occurred in wetter soils.

Keywords: Antarctica, hypolith, soil, dry valleys, geochip, functional diversity, stress response

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Authors: Irit Rutman Halili, Tehila Zvulun, Natali Elgabsi, Revaya Cohen, Shlomo Sarig

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Active sand dunes (ASD) may cause significant damage to field crops and livelihood, and therefore, it is necessary to find a treatment that would enhance ADS soil stability. Biological soil crusts (biocrusts) contain microorganisms on the soil surface. Metabolic polysaccharides secreted by biocrust cyanobacteria glue the soil particles into aggregates, thereby stabilizing the soil surface. Filter cake powder (FCP) is a waste by-product in the final stages of the production of sugar from sugarcane, and its disposal causes significant environmental pollution. FCP contains high concentrations of polysaccharides and has recently been shown to be soil stability enhancing agent in ASD. It has been reported that adding FCP to the ASD soil surface by dispersal significantly increases the level of penetration resistance of soil biocrust (PRSB) nine weeks after a single treatment. However, it was not known whether a similar effect could be obtained by administering the FCP in liquid form by means of spraying. It has now been found that spraying a water solution of FCP onto the ASD soil surface significantly increased the level of penetration resistance of soil biocrust (PRSB) three weeks after a single treatment. These results suggest that FCP spraying can be used as a short-term soil stability-enhancing agent for ASD, while administration by dispersal might be more efficient over the long term. Finally, an additional benefit of using FCP as a soil stabilizer, either by dispersal or by spraying, is the reduction in environmental pollution that would otherwise result from the disposal of FCP solid waste.

Keywords: active sand dunes, filter cake powder, biological soil crusts, penetration resistance of soil biocrust

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Authors: Medina Kadiri

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Blue-green otherwise called cyanobacteria, produce an array of biotoxins grouped into five categories notably hapatotoxins, neurotoxins, cytotoxins, dermatotoxins, and irritant toxins. Microcystins which are examples of hepatotoxins produced by blue-green algae Microcystins comprise the most common group of the cyanobacterial toxins. Blue-green algae flourish in aquatic environments, whether marine, brackish or freshwater, producing blooms in different forms such as microscopic, mats, or unsightly odoriferous scums. Microcystins biotoxins cause a plethora of animal and human hazards such as liver damage/cirrhosis and cancer, kidney damage, dermatitis, tinnitus, gastroenteritis, sore throat, nausea, myalgia, neurological problems, respiratory irritation and death. Water samples were collected from coastal regions of Nigeria in March 2014, June 2014, October 2014 and January 2015 and analyzed with Enzyme Linked Immunosorbent Assay (ELISA) kits. Microcystin biotoxin was recorded in all sites both during dry and wet seasons. The range of microcystins found was 0.000041-There was a seasonal trend of increasing microcystin concentrations from March till Octobers and a decrease thereafter. Generally in the oceanic waters, microcystin levels were highest at Cross Rivers in March and January, Barbeach in June and Lekki in October. In the adjoining riverine ecosystems, on the other hand, the highest concentrations of microcystin were observed at Akwa Ibom in March, June and October and in Bayelsa in January. Continuous monitoring and screening of coastal water bodies is suggested to minimize the health risks of cyanobacterial biotoxins to coastal communities of Nigeria.

Keywords: biotoxins, harmful algae, marine, microcystin, Nigeria

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Authors: Geno Paul S. Cumla, Jan Mariel M. Gentiles, M. Brenda Gajelan-Samson

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Eutrophication is a process where in there is a surplus of nutrients present in a lake or river. Harmful cyanobacteria, hypoxia, and primarily algae, which contain toxins, grow because of the excess nutrients. Algal blooms can cause fish kills, limiting the light penetration which reduces growth of aquatic organisms, causing die-offs of plants and produce conditions that are dangerous to aquatic and human life. The main cause for eutrophication is the presence of excessive amounts of phosphorus (P) and nitrogen (N). Nitrogen is necessary for the production of the plant tissues and is usually used to synthesize proteins. Nitrate is a compound that contains nitrogen, and at elevated levels it can cause harmful effects. Excessive amounts of phosphorus, displaced through human activity, is the major cause of algae growth and as well as degraded water quality. To accomplish this study the Assessment of Soluble inorganic nitrogen (SIN), Assessment of Soluble reactive phosphate (SRP), Determination of Chlorophyll a (Chl-a) concentration, and Determination of Dominating Taxa were done. The study addresses the high probability of algal blooms in Maqueda Bay by assessing the biotic and abiotic factors of Antiao and Jiabong rivers. The data predicts the overgrowth of algae and to create awareness to prevent the event from taking place. The study assesses the adverse effects that could be prevented by understanding and controlling algae. This should predict future cases of algal blooms and allow government agencies which require data to create programs to prevent and assess these issues.

Keywords: eutrophication, chlorophyll a, nitrogen, phosphorus, red tide, Kjeldahl method, spectrophotometer, assessment of soluble inorganic nitrogen, SIN, assessment of soluble reactive phosphate, SRP

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Authors: Nancy Awuor Oduor, Nils Moosdorf

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Harmful Algal Blooms (HABs) are a threat to coastal water quality, marine biodiversity, and human health. The attention on HABs and associated phycotoxins is still very low in tropical coastal developing countries despite the high dependence of local communities on coastal and marine resources for food and livelihoods and the growing evidence of the global increase in HABs frequency, toxicity, and geographical expansion. Lack of HABs monitoring thus creates a high risk of exposure due to uncertainty. This study assessed the spatial and temporal variability and effects of potential HAB-forming species in Kenya’s coastal waters. The preliminary results from 463 sampled collected over a series of 10 coastal surveys conducted over 267 Km of Kenya’s coastline between August 2021 and July 2022 revealed the presence of 87 potential algal blooming species belonging to 47 genera dominated by species capable of producing toxins, causing physical harm and high biomass at 41, 31 and 21 % respectively. The taxonomic composition was also dominated by dinoflagellates at 47%, followed by diatoms, cyanobacteria, and silicoflagellates at 39, 12, and 2%, respectively. About 92 % of the toxin-producing species were established in the creek waters. However, there were no significant variations established in species richness between the dry and wet seasons. Paralytic Shellfish Poisoning (PSP) toxin-producing dinoflagellates Alexandrium spp., Aphanizomenon spp., Gonyaulax spp., Gymnodinium spp., and Brachydinium capitatum, and Amnesic Shellfish Poisoning (ASP) Toxin producing diatoms Amphora spp., Nitzschia spp. and Pseudo-nitzschia spp. Frequented the area in low cell densities ranging between 5 and 1500 cells/L. However, no domoic acid (DA) and saxitoxins (SXTs) were detected during the July surveys. This does not mean that the toxins are absent in the area, and longer studies are recommended.

Keywords: harmful algal blooms, phycotoxins, saxitoxin, domoic acid, Kenya

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Authors: Velasco-Rendón María Del Carmen, Cuéllar-Bermúdez Sara Paulina, Parra-Saldívar Roberto

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Phycocyanin is a blue pigment protein with fluorescent activity produced by cyanobacteria. It has been recently studied to determine its anticancer, antioxidant and antiinflamatory potential. Since 2014 it was approved as a Generally Recognized As Safe (GRAS) proteic pigment for the food industry. Therefore, phycocyanin shows potential for the food, nutraceutical, pharmaceutical and diagnostics industry. Conventional phycocyanin extraction includes buffer solutions and ammonium sulphate followed by chromatography or ATPS for protein separation. Therefore, further purification steps are time-requiring, energy intensive and not suitable for scale-up processing. This work presents an alternative to conventional methods that also allows large scale application with commercially available equipment. The extraction was performed by exposing the dry biomass to mechanical cavitation and salting out with NaCl to use an edible reagent. Also, isoelectric point precipitation was used by addition of HCl and neutralization with NaOH. The results were measured and compared in phycocyanin concentration, purity and extraction yield. Results showed that the best extraction condition was the extraction by salting out with 0.20 M NaCl after 30 minutes cavitation, with a concentration in the supernatant of 2.22 mg/ml, a purity of 3.28 and recovery from crude extract of 81.27%. Mechanical cavitation presumably increased the solvent-biomass contact, making the crude extract visibly dark blue after centrifugation. Compared to other systems, our process has less purification steps, similar concentrations in the phycocyanin-rich fraction and higher purity. The contaminants present in our process edible NaCl or low pHs that can be neutralized. It also can be adapted to a semi-continuous process with commercially available equipment. This characteristics make this process an appealing alternative for phycocyanin extraction as a pigment for the food industry.

Keywords: extraction, phycocyanin, precipitation, scale-up

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Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

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Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

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Authors: Apurva Gupta, Surendra Singh

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Spirulina spp., as a promising source of many commercially valuable products, is grown photo autotrophically in open ponds and raceways on a large scale. However, the economic exploitation in an open system seems to have been limited because of lack of multiple stress-tolerant strains. The present study aims to isolate a stable stress tolerant mutant of Spirulina platensis with improved growth rate and enhanced potential to produce its commercially valuable bioactive compounds. N-methyl-n'-nitro-n-nitrosoguanidine (NTG) at 250 μg/mL (concentration permitted 1% survival) was employed for chemical mutagenesis to generate random mutants and screened against NaCl. In a preliminary experiment, wild type S. platensis was treated with NaCl concentrations from 0.5-1.5 M to calculate its LC₅₀. Mutagenized colonies were then screened for tolerance at 0.8 M NaCl (LC₅₀), and the surviving colonies were designated as NaCl tolerant mutants of S. platensis. The mutant cells exhibited 1.5 times improved growth against NaCl stress as compared to the wild type strain in control conditions. This might be due to the ability of the mutant cells to protect its metabolic machinery against inhibitory effects of salt stress. Salt stress is known to adversely affect the rate of photosynthesis in cyanobacteria by causing degradation of the pigments. Interestingly, the mutant cells were able to protect its photosynthetic machinery and exhibited 4.23 and 1.72 times enhanced accumulation of Chl a and phycobiliproteins, respectively, which resulted in enhanced rate of photosynthesis (2.43 times) and respiration (1.38 times) against salt stress. Phycocyanin production in mutant cells was observed to enhance by 1.63 fold. Nitrogen metabolism plays a vital role in conferring halotolerance to cyanobacterial cells by influx of nitrate and efflux of Na+ ions from the cell. The NaCl tolerant mutant cells took up 2.29 times more nitrate as compared to the wild type and efficiently reduce it. Nitrate reductase and nitrite reductase activity in the mutant cells also improved by 2.45 and 2.31 times, respectively against salt stress. From these preliminary results, it could be deduced that enhanced nitrogen uptake and its efficient reduction might be a reason for adaptive and halotolerant behavior of the S. platensis mutant cells. Also, the NaCl tolerant mutant of S. platensis with significant improved growth and phycocyanin accumulation compared to the wild type can be commercially promising.

Keywords: chemical mutagenesis, NaCl tolerant mutant, nitrogen metabolism, photosynthetic machinery, phycocyanin

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Authors: Gabriela Kalcikova, Igor Bosevski, Ula Rozman, Andreja Zgajnar Gotvajn

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Antibiotics are extensively used in human medicine and also in animal husbandry to prevent or control infections. Recently, a lot of attention has been put on veterinary antibiotics, because their global consumption is increasing and it is expected to be 106.600 tons in 2030. Most of veterinary antibiotics are introduced into the environment via animal manure, which is used as fertilizer. One of such veterinary antibiotics is tiamulin. It is used the form of fumarate for treatment of pig and poultry. It is used against prophylaxis of dysentery, pneumonia and mycroplasmal infections, but its environmental impact is practically unknown. Tiamulin has been found very persistent in animal manure and thus it is expected that can be, during rainfalls, transported into the aquatic environment and affect various organisms. For assessment of its environmental impact, it is necessary to evaluate its biodegradability and toxicity to various organisms from different levels of a food chain. Therefore, the aim of our study was to evaluate ready biodegradability and toxicity of tiamulin fumarate to various organisms. Bioassay used included luminescent bacterium Vibrio fischeri heterotrophic and nitrifying microorganisms of activated sludge, water flea Daphnia magna and duckweed Lemna minor. For each species, EC₅₀ values were calculated. Biodegradability test was used for determination of ready biodegradability and it provides information about biodegradability of tiamulin under the most common environmental conditions. Results of our study showed that tiamulin differently affects selected organisms. The most sensitive organisms were water fleas with 48hEC₅₀ = 14.2 ± 4.8 mg/L and duckweed with 168hEC₅₀ = 22.6 ± 0.8 mg/L. Higher concentrations of tiamulin (from 10 mg/L) significantly affected photosynthetic pigments content in duckweed and concentrations above 80 mg/L cause visible chlorosis. It is in agreement with previous studies showing significant effect of tiamulin on green algae and cyanobacteria. Tiamuline has a low effect on microorganisms. The lower toxicity was observed for heterotrophic microorganisms (30minEC₅₀ = 1656 ± 296 mg/L), than Vibrio fisheri (30minEC₅₀ = 492 ± 21) and the most sensitive organisms were nitrifying microorganisms (30minEC₅₀ = 183 ± 127 mg/L). The reason is most probably the mode of action of tiamulin being effective to gram-positive bacteria while gram-negative (e.g., Vibrio fisheri) are more tolerant to tiamulin. Biodegradation of tiamulin was very slow with a long lag-phase being 20 days. The maximal degradation reached 40 ± 2 % in 43 days of the test and tiamulin as other antibiotics (e.g. ciprofloxacin) are not easily biodegradable. Tiamulin is widely used antibiotic in veterinary medicine and thus present in the environment. According to our results, tiamulin can have negative effect on water fleas and duckweeds, but the concentrations are several magnitudes higher than that found in any environmental compartment. Tiamulin is low toxic to tested microorganisms, but it is very low biodegradable and thus possibly persistent in the environment.

Keywords: antibiotics, biodegradability, tiamulin, toxicity

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Authors: Irina M. Kolesnikova, Andrey M. Gaponov, Sergey A. Roumiantsev, Tatiana V. Grigoryeva, Dilyara R. Khusnutdinova, Dilyara R. Kamaldinova, Alexander V. Shestopalov

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Background. Obese patients have unequal risks of metabolic disorders. It is accepted to distinguish between metabolically healthy obesity (MHO) and metabolically unhealthy obesity (MUHO). MUHO patients have a high risk of metabolic disorders, insulin resistance, and diabetes mellitus. Among the other things, the gut microbiota also contributes to the development of metabolic disorders in obesity. Obesity is accompanied by significant changes in the gut microbial community. In turn, bacterial translocation from the intestine is the basis for the blood microbiome formation. The aim was to study the features of the blood microbiome in patients with various metabolic types of obesity. Patients, materials, methods. The study included 116 healthy donors and 101 obese patients. Depending on the metabolic type of obesity, the obese patients were divided into subgroups with MHO (n=36) and MUHO (n=53). Quantitative and qualitative assessment of the blood microbiome was based on metagenomic analysis. Blood samples were used to isolate DNA and perform sequencing of the variable v3-v4 region of the 16S rRNA gene. Alpha diversity indices (Simpson index, Shannon index, Chao1 index, phylogenetic diversity, the number of observed operational taxonomic units) were calculated. Moreover, we compared taxa (phyla, classes, orders, and families) in terms of isolation frequency and the taxon share in the total bacterial DNA pool between different patient groups. Results. In patients with MHO, the characteristics of the alpha-diversity of the blood microbiome were like those of healthy donors. However, MUHO was associated with an increase in all diversity indices. The main phyla of the blood microbiome were Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. Cyanobacteria, TM7, Thermi, Verrucomicrobia, Chloroflexi, Acidobacteria, Planctomycetes, Gemmatimonadetes, and Tenericutes were found to be less significant phyla of the blood microbiome. Phyla Acidobacteria, TM7, and Verrucomicrobia were more often isolated in blood samples of patients with MUHO compared with healthy donors. Obese patients had a decrease in some taxonomic ranks (Bacilli, Caulobacteraceae, Barnesiellaceae, Rikenellaceae, Williamsiaceae). These changes appear to be related to the increased diversity of the blood microbiome observed in obesity. An increase of Lachnospiraceae, Succinivibrionaceae, Prevotellaceae, and S24-7 was noted for MUHO patients, which, apparently, is explained by a magnification in intestinal permeability. Conclusion. Blood microbiome differs in obese patients and healthy donors at class, order, and family levels. Moreover, the nature of the changes is determined by the metabolic type of obesity. MUHO linked to increased diversity of the blood microbiome. This appears to be due to increased microbial translocation from the intestine and non-intestinal sources.

Keywords: blood microbiome, blood bacterial DNA, obesity, metabolically healthy obesity, metabolically unhealthy obesity

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Authors: C. J. Tien, C. S. Chen, S. F. Huang, Z. X. Wang

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Algae in soil ecosystems can produce organic matters and oxygen by photosynthesis. Heterocyst-forming cyanobacteria can fix nitrogen to increase soil nitrogen contents. Secretion of mucilage by some algae increases the soil water content and soil aggregation. These actions will improve soil quality and fertility, and further increase abundance and diversity of soil microorganisms. In addition, some mixotrophic and heterotrophic algae are able to degrade petroleum hydrocarbons. Therefore, the objectives of this study were to analyze the effects of algal addition on the degradation of total petroleum hydrocarbons (TPH), diversity and activity of bacteria and algae in the diesel-contaminated soil under different nutrient contents and frequency of plowing and irrigation in order to assess the potential bioremediation technique using edaphic algae. The known amount of diesel was added into the farmland soil. This diesel-contaminated soil was subject to five settings, experiment-1 with algal addition by plowing and irrigation every two weeks, experiment-2 with algal addition by plowing and irrigation every four weeks, experiment-3 with algal and nutrient addition by plowing and irrigation every two weeks, experiment-4 with algal and nutrient addition by plowing and irrigation every four weeks, and the control without algal addition. Soil samples were taken every two weeks to analyze TPH concentrations, diversity of bacteria and algae, and catabolic genes encoding functional degrading enzymes. The results show that the TPH removal rates of five settings after the two-month experimental period were in the order: experiment-2 > expermient-4 > experiment-3 > experiment-1 > control. It indicated that algal addition enhanced the degradation of TPH in the diesel-contaminated soil, but not for nutrient addition. Plowing and irrigation every four weeks resulted in more TPH removal than that every two weeks. The banding patterns of denaturing gradient gel electrophoresis (DGGE) revealed an increase in diversity of bacteria and algae after algal addition. Three petroleum hydrocarbon-degrading algae (Anabaena sp., Oscillatoria sp. and Nostoc sp.) and two added algal strains (Leptolyngbya sp. and Synechococcus sp.) were sequenced from DGGE prominent bands. The four hydrocarbon-degrading bacteria Gordonia sp., Mycobacterium sp., Rodococcus sp. and Alcanivorax sp. were abundant in the treated soils. These results suggested that growth of indigenous bacteria and algae were improved after adding edaphic algae. Real-time polymerase chain reaction results showed that relative amounts of four catabolic genes encoding catechol 2, 3-dioxygenase, toluene monooxygenase, xylene monooxygenase and phenol monooxygenase were appeared and expressed in the treated soil. The addition of algae increased the expression of these genes at the end of experiments to biodegrade petroleum hydrocarbons. This study demonstrated that edaphic algae were suitable biomaterials for bioremediating diesel-contaminated soils with plowing and irrigation every four weeks.

Keywords: catabolic gene, diesel, diversity, edaphic algae

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Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

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The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

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Authors: Marcin Debowski, Marcin Zielinski, Miroslaw Krzemieniewski, Agata Glowacka-Gil, Paulina Rusanowska, Magdalena Zielinska, Agnieszka Cydzik-Kwiatkowska

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Technologies of biogas upgrading are now perceived as competitive solution combustion and production of electricity and heat. Biomethane production will ensure broader application as energy carrier than biogas. Biomethane can be used as fuel in internal combustion engines or introduced into the natural gas transmission network. Therefore, there is a need to search for innovative, economically and technically justified methods for biogas enrichment. The aim of this paper is to present a technology solution for biogas upgrading with immobilized algae biomass. Reactor for biogas upgrading with immobilized algae biomass can be used for removing CO₂ from the biogas, flue gases and the waste gases especially coming from different industry sectors, e.g. from the food industry from yeast production process, biogas production systems, liquid and gaseous fuels combustion systems, hydrocarbon processing technology. The basis for the technological assumptions of presented technology were laboratory works and analyses that tested technological variants of biogas upgrading. The enrichment of biogas with a methane content of 90-97% pointed to technological assumptions for installation on a technical scale. Reactor for biogas upgrading with algae biomass is characterized by a significantly lower cubature in relation to the currently used solutions which use CO₂ removal processes. The invention, by its structure, assumes achieving a very high concentration of biomass of algae through its immobilization in capsules. This eliminates the phenomenon of lowering the pH value, i.e. acidification of the environment in which algae grow, resulting from the introduction of waste gases at a high CO₂ concentration. The system for introducing light into algae capsules is characterized by a higher degree of its use, due to lower losses resulting from the phenomenon of absorption of light energy by water. The light from the light source is continuously supplied to the formed biomass of algae or cyanobacteria in capsules by the light tubes. The light source may be sunlight or a light generator of a different wavelength of light from 300 nm to 800 nm. A portion of gas containing CO₂, accumulated in the tank and conveyed by the pump is periodically introduced into the housing of the photobioreactor tank. When conveying the gas that contains CO₂, it penetrates the algal biomass in capsules through the outer envelope, displacing, from the algal biomass, gaseous metabolic products which are discharged by the outlet duct for gases. It contributes to eliminating the negative impact of this factor on CO₂ binding processes. As a result of the cyclic dosing of gases containing carbon dioxide, gaseous metabolic products of algae are displaced and removed outside the technological system. Technology for biogas upgrading with immobilized algae biomass is suitable for the small biogas plant. The advantages of this technology are high efficiency as well as useful algae biomass which can be used mainly as animal feed, fertilizers and in the power industry. The construction of the device allows effective removal of carbon dioxide from gases at a high CO₂ concentration.

Keywords: biogas, carbon dioxide, immobilised biomass, microalgae, upgrading

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: Shewit Kidane, Abebe Getahun, Wassie Anteneh, Admassu Demeke, Peter Goethals

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We have reviewed major findings of scientific studies on Lake Tana fish resources and their threats. The aim was to provide summarized information for all concerned bodies and international readers to get full and comprehensive picture about the lake’s fish resource and conservation problems. The Lake Tana watershed comprise 28 fish species, of which 21 are endemic. Moreover, Lake Tana is the one among the top 250 lake regions of global importance for biodiversity and it is world recognized migratory birds wintering site. Lake Tana together with its adjacent wetlands provide directly and indirectly a livelihood for more than 500,000 people. However, owing to anthropogenic activities, the lake ecosystem as well as fish and attributes of the fisheries sector are severely degraded. Fish species in Lake Tana are suffering due to illegal fishing, damming, habitat/breeding ground degradation, wastewater disposal, introduction of exotic species, and lack of implementing fisheries regulations. Currently, more than 98% of fishers in Lake Tana are using the most destructive monofilament. Indeed, dams, irrigation schemes and hydropower are constructed in response to the emerging development need only. Mitigation techniques such as construction of fish ladders for the migratory fishes are the most forgotten. In addition, water resource developers are likely unaware of both the importance of the fisheries and the impact of dam construction on fish. As a result, the biodiversity issue is often missed. Besides, Lake Tana wetlands, which play vital role to sustain biodiversity, are not wisely utilised in the sense of the Ramsar Convention’s definition. Wetlands are considered as unhealthy and hence wetland conversion for the purpose of recession agriculture is still seen as advanced mode of development. As a result, many wetlands in the lake watershed are shrinking drastically over time and Cyprus papyrus, one of the characteristic features of Lake Tana, has dramatically declined in its distribution with some local extinction. Furthermore, the recently introduced water hyacinth (Eichhornia crassipes) is creating immense problems on the lake ecosystem. Moreover, currently, 1.56 million tons of sediment have deposited into the lake each year and wastes from the industries and residents are directly discharged into the lake without treatment. Recently, sign of eutrophication is revealed in Lake Tana and most coarsely, the incidence of cyanobacteria genus Microcystis was reported from the Bahir Dar Gulf of Lake Tana. Thus, the direct dependency of the communities on the lake water for drinking as well as to wash their body and clothes and its fisheries make the problem worst. Indeed, since it is home to many endemic migratory fish, such kind of unregulated developmental activities could be detrimental to their stocks. This can be best illustrated by the drastic stock reduction (>75% in biomass) of the world unique Labeobarbus species. So, unless proper management is put in place, the anthropogenic impacts can jeopardize the aquatic ecosystems. Therefore, in order to sustainably use the aquatic resources and fulfil the needs of the local people, every developmental activity and resource utilization should be carried out adhering to the available policies.

Keywords: anthropogenic impacts, dams, endemic fish, wetland degradation

Procedia PDF Downloads 208