Production of Spirulina platensis Using Inexpensive Local Resources in Palestine

Spirulina platensis is a special and unique cyanobacteria that is produced worldwide with a varied cost of cultivation media. In this study, five main experiments with different treatments were performed to evaluate the possibility of using cheap aquaculture water for Spirulina production, to test if solutions made by plant ash (PAS) could be used for Spirulina production, to determine if brackish water (BW) and mining water have a good impact on Spirulina production, to create a medium composed of cheap chemicals and fertilizers to be used for Spirulina cultivation, and to test if a mixture made from local components could be used to produce Spirulina. All experiments were performed via growth and dry weight measurements, including determination of chemical and physical characteristics of the samples with a comparison with Zarrouk medium (ZM) as a reference for each experiment, and all experiments were performed for 21 days to determine the best media type that lasts longer for commercial purposes. In all experiments, pH values were between 8 and 11, and EC was between 9.8 and 30 ms/cm, while temperature was at 30°C and 35°C, and light was at 1500 and 5000 Lux for 16 h light and 8 h dark. Spirulina can grow in (FW). It can also grow in FW diluted with BW. Also a 3% PAS can be used as a source to cultivate Spirulina at a very low price compared to ZM. The chemical fertilizer formula was one of the best types among all treatments with a good price. A mixture of these local resources could be a very good cheap alternative source. The main result that was obtained from all experiments in this study is the ability of Spirulina to grow within a wide range of chemical parameters at a lower price.

Investigation on the Detrimental Effects of Salt Stress on Photosynthesis of Spirulina platensis

[Objective] This study aimed to gain better understanding of the effects of salt stress on photosynthesis of Spirulina platensis. [Method] A salt stress simulation experiment was carried out, in which Spirulina platensis cells were incubated with different salt concentrations （0, 0.1, 0.3, 0.5, 0.8, and 1.0 mol/L of NaCI）. Subsequently, some photosynthesis-related parameters were determined after incubated at various NaCI concentrations for 24 h. [Result] Under our experimental conditions, chlorophyll, carotenoid, phycocyanin and allophycocyanin contents, and photosynthesis rate decreased with the increasing NaCI concentrations. Above the 0.5 mol/L Na- CI level, the detrimental effect of salt stress became more severe. [Conclusion] From the results obtained in this investigation, we can conclude that the decrease in chlorophyll, carotenoid, phycocyanin and allophycocyanin contents may be the most important reason causing the decline in photosynthsis rate which strongly affects the biomass yield of Spirulina platensis in outdoor cultivation

Comparison of the therapeutic effects of extracts from Spirulina platensis and amnion membrane on inflammation-associated corneal neovascularization

AIM: To compare the therapeutic effects of polysaccharide extract from Spirulina platensis (PSP) and extract from amnion membrane CAME) on alkali burn-induced corneal neovascularization (CorNV). METHODS: PSP and AME were Extracted from dry powder of Spirulina platensis and human aminion membrane respectively. Murine CorNV was induced by applying 1N sodiumhydroxide (NaOH) solution directly on the mice corneas. PSP and AME extracts were administered topically on the corneas 4 times daily for 7 days. The therapy effects of PSP and AME extracts were evaluated daily using slit-lamp. At the end of the therapy, corneas were harvested for H&E staining, masson trichrome staining, immunohistochemical study, and semi-quantification reverse transcriptive PCR (RT-PCR) was utilized for measurement of inflammation-related molecules. RESULTS: Topical application of PSP extract had significant therapeutic effects on CorNV that could be shown in various assays of the corneas. Compared with AME Extract, PSP extract treatment was more effective in suppressing CorNV in berms of vessel length and levels of cluster of differentiation 31 (CD31) proteins or the angiogenesis related genes like vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9). PSP also inhibited inflammation more markedly by more effectively inhibiting mononuclear and polymorphonuclear cells infiltration into the corneal stroma and reducing levels of stromal cell-derived factor-1 (SDF1), tumor necrosis factor-alpha (TNF alpha) and macrophage inflammatory protein-3 (MIP3a). In addition, corneas of PSP group had a more regular and compact architecture of collagen with thinner corneal thickness than in the AME group. ' CONCLUSION: Polysaccharide extract from Spirulina platensis inhibited alkali burn-induced inflammation and CorNV more effectively than AME Extract at the studied doses, thus may be used for the therapy of corneal diseases involving neovascularization and inflammation.

Effects of 50 Percent Substitution of Soybean Meal by Alternative Proteins from <i>Hermetia illucens</i>or Spirulina platensis in Meat-Type Chicken Diets with Graded Amino Acid Supply

Alternative protein sources, such as insects or algae meals are in special focus of animal nutrition in order to replace soybean meal (SBM). As part of the multidisciplinary project “sustainability transitions” this study evaluated effects of replacing SBM by partly defatted larvae meal from the black soldier fly, Hermetia illucens or meal from the micro algae Spirulina platensis in broiler diets. The aim of the current study was to investigate the chickens’ growth performance and the intestinal morphology as well as the health status. 288 one-day-old male growing chickens (Ross 308) from a commercial hatchery were randomly allotted to 48 pens (6 birds per pen) for the growth study with five diets and feed supply on free choice level. The control diet was based on wheat, corn and SBM. The experimental diets replaced 50% of SBM by the alternative proteins under study, both on a basic level of amino acid (AA) supplementation (Lys and Met added equal to the control diet) (diet HM and SM) and an extended level of AA fortification (Lys, Met, Thr, Arg, Val added) (diet HM+ and SM+). Response of chickens was evaluated by zoo-technical parameters, which were under weekly control (e.g. growth, feed intake, feed conversion ratio, mortality). After finishing the growth study birds from control and the experimental diets at the basic level of AA supplementation were slaughtered after 12 hour fastening and utilized for gut morphometric analysis and histological evaluation of the health status. Diets at a basic level of AA fortification led to significant depression of growth, feed intake, feed conversion ratio, and protein conversion ratio, respectively. In addition, the acceptance of the Spirulina diet was lower (p ≤ 0.001) as compared to the Hermetia based diet. However, the extended level of AA supplementation improved all parameters significantly. Diets without extended AA supply altered some morphological parameters of the intestinal wall, but the nutritional significance of this observation needs to be verified in AA balanced diets. The health state of chicken was not impaired by the diets under study.

ISOLATION OF PLASMID FROM THE BLUE-GREEN ALGA SPIRULINA PLATENSIS

CCC plasmid was isolated from an economically important blue-green alga- Spirulina platensis (1.7×106 dalton from the S6 strain and 1.2×106 dalton from the F, strain) using a rapid method based on ultrasonic disruption of algal cells and alkaline removal of chromosomal DNA. The difference in the molecular weight of the OOC DNAs from the two strains differing in form suggests that plasmid may be related with the differentiation of algal form. This modified method, which does not use any lysozyme, is a quick and effective method of plasmid isolation, especially for filamentous blue-green algae.

Science Letters:Culture of Spirulina platensis in human urine for biomass production and O_2 evolution

Attempts were made to culture Spirulina platensis in human urine directly to achieve biomass production and O2 evolution, for potential application to nutrient regeneration and air revitalization in life support system. The culture results showed that Spirulinaplatensis grows successfully in diluted human urine, and yields maximal biomass at urine dilution ratios of 140-240. Accumulation of lipid and decreasing of protein occurred due to N deficiency. O2 release rate of Spirulina platensis in diluted human urine was higher than that in Zarrouk medium.

Effects of N source and nitrification pretreatment on growth of Arthrospira platensis in human urine

Culture ofArthrospiraplatensis （Spirulinaplatens） in human urine was investigated to get valuable biomass. NO3-N was the proper N source, in comparison with other N source, including urea, NH4-N and NO2-N. As a result, aerobic nitrification of human urine was performed, with above 93.6% nitrification percentage finally achieved with total-N （TN） load of 46.52 mg/（L.d）, in which Arthrospira platensis was successfully grown. The main compositions of the obtained biomass are close to those in Zarrouk medium. Thus, it is possible to culture Arthrospiraplatensis in nitrified human urine for food production within bioregenerative life support systems （BLSSs）.

Protective effects of a polysaccharide from Spirulina platensis on dopaminergic neurons in an MPT-Pinduced Parkinson's disease model in C57BL/6J mice

The present study aimed to determine whether a polysaccharide obtained fromSpirulina platensis shows protective effects on dopaminergic neurons. A Parkinson’s disease model was established through the intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyr-idine （MPTP） in C57BL/6J mice. Prior to the MPTP injection, some mice were pretreated with intraperitoneal injections of a polysaccharide derived fromSpirulina platensis once daily for 10 days. The results showed that the immunoreactive staining and mRNA expression of the dopa-mine transporter and tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, in the substantia nigra, were signiifcantly increased in mice pretreated with 800 mg/kg of the poly-saccharide compared with those in MPTP-treated mice. The activities of superoxide dismutase and glutathione peroxidase in the serum and midbrain were also increased signiifcantly in mice injected with MPTP after pretreatment with the polysaccharide fromSpirulina platensis. By con-trast, the activity of monoamine oxidase B in serum and midbrain maintained unchanged. These experimental ifndings indicate that the polysaccharide obtained fromSpirulina platensis plays a protective role against the MPTP-induced loss of dopaminergic neurons in C57BL/6J mice, and that the antioxidative properties of this polysaccharide likely underlie its neuroprotective effect.

Effect of CeCl_3 on the Photosynthetic Characteristics of Spirulina Platensis

In this paper we report that CeCl3 could accelerate the synthesis of chlorophyll a and protein in spirulina platensis. and enhance the activity of oxygen evolution. We also found that the accelerations of CeCl3 in spirulina platensis culture solution of low NaHCO3 were more than in culture solution of high NaHCO3, therefore it was related to the concentration of NaHCO3 The absorption spectrum and low temperature (77 K) fluorescence emission spectrum of spirulina platensis with CeCl3 treated were significantly different from their controls,i.e.its absorption peak of phycocyanin at 620 nm was higher than that of the control.its absorption peak at the long wave region migrated from 676 to 680 nm, and its fluorecence emission ratios (730 nm/685～695 nm) of photosystem I(PSI) to photosystem Ⅱ(PS Ⅱ) were also higher than those of the control; the pitch of the spiral algal body became shorter, and induced structural changes too.

Bioaccumulation of Pb^(2+) and Its Effects on Growth,Morphology and Pigment Contents of Spirulina (Arthrospira) platensis

A laboratory experiment was conducted to assess the bioaccumulation of Pb^2＋ and its effects on growth, morphology and pigment contents of Spirulina （Arthrosl）ira）platensis. The specimen cultured in Zarrouk liquid medium was treated with various initial metal concentrations （0, 5, 10, 30, 50 and 10μgmL^-1）. The growth ofS. platensis was adversely affected by Pb〉 at high concentrations （30, 50 and 100μgmL^-1）. However, at low concentrations （5μgmL^-1）, Pb^2＋ could stimulate its growth slightly. The pigment contents （chlorophyllα and β carotene） were decreased in a dose-dependent manner. The highest reductions （67% and 53% respectively in chlorophyll α andβ carotene） were observed in 100 μg mL^-1 treatment group. The LC50 （96 h） of Pb^2- was measured as 75.34 μg mL^-1. Apart from a few cases of filament breakages at elevated concentrations （50 and 100 μgmL^-1）, morphological abnormalities are not specific. Metal bioaccumulation increased with Pb^2- concentrations, but decreased with exposure time. The maximum accumulated amount was 188 mgg^-1 dry weight. The bioconcentration factor （BCF） reached to a peak at day 2, followed by a gradual reduction tbr all the exposure concentrations. S. platensis is able to tolerate considerably high Pb^2＋ concentrations. Consequently it can be used as a potential species to remove heavy metal from contaminated waters.

Bi-Enzymatic Conductometric Biosensor for Detection of Heavy Metal Ions and Pesticides in Water Samples Based on Enzymatic Inhibition in <i>Arthrospira platensis</i>

An original bi-enzymatic biosensor was designed by immobilizing Arthrospira platensis cells, called Spirulina, on gold interdigitated transducers. Phosphatase and esterase activities were inhibited, respectively, by heavy metals and by pesticides. Inhibition activities were observed with different mixtures of pesticides + heavy metals. The quantification limits for Cd2+ and Hg2+ are 10-20 M in mixture and in pure solution. The quantification limits of parathion-methyl, paraoxon-methyl and triazine are respectively 10-20 M, 10-18 M and 10-20 M in mixture and pure solutions. These results show that there is no synergistic effect between the two families of pollutants. Qualitative contamination of effluent samples and their purification after passing through a municipal wastewater treatment plant were observed by our bi-enzymatic biosensor and confirmed by classical analytical techniques for heavy metal ions.

Improving the Dietary Protein Quality by Amino Acid Fortification with a High Inclusion Level of Micro Algae (<i>Spirulina platensis</i>) or Insect Meal (<i>Hermetia illucens</i>) in Meat Type Chicken Diets

The objective of this study was to measure protein quality parameters (PPV, NPU and NPUstd) of chicken diets with complete substitution of soybean meal (SBM) by Spirulina meal (SM) or partly defatted Hermetia meal (HM). N balance experiments were based on the quantitative excreta collection method, divided into starter period (10 - 20 d) and grower period (25 - 35 d). The study utilized 70 all male meat type chicken (Ross 308). Data assessment applied the exponential N utilization model of the “Goettingen approach”. The control diet was based on wheat, corn and SBM. In four experimental diets SBM was completely substituted by SM or HM, but fortified with feed amino acids (AA) both on a basic level of supplementation (Lys and Met added equal to the control diet) and on an extended level (Lys, Met, Thr, Arg, Val, Ile, His added). At a basic level of AA supplementation, complete replacement of SBM by SM or HM in chicken diets depressed dietary protein quality significantly (p 0.05). However, the extended level of AA supplementation improved protein quality parameters of the diets with both of the alternative proteins significantly (p < 0.05), but still generally not on par with the control diet. The observed responses were accentuated when the well-known effect of N intake on protein utilization was eliminated through the standardization of N intake by application of the “Goettingen approach”.

Effect of Supplementation of Spirulina platensis to Enhance the Zinc Status in Plants of Amaranthus gangeticus, Phaseolus aureus and Tomato

Trace elements or micronutrients play a major role in the metabolism of both plants and humans. Zinc has a major specific role in metabolism when compared with other elements. The microbial biofertilizers are applied in the form of seaweed liquid extracts, microbial inoculants, biostimulators and biofortification agents. All these categories of microbial biofertilizers are involved in the enhancement of plant nutrient uptake. In the present study, Spirulina platensis is used as a biofortification agent to enhance zinc levels in cultivars of Amaranthus gangeticus, Phaseolus aureus and Tomato. Different experimental methods were followed including soaking seeds in different concentrations of Spirulina (5, 10, 15, 20, 25 and 30 g in 100 ml of water);soaking seeds in Spirulina hydrolysate at different time intervals (1, 2, 3, 4, 5 hrs and overnight);Spirulina in combination with biofertilizers, chemical fertilizer, organic fertilizer and vermicompost in various proportions (25:75;50:50;75:25) and foliar spray with different concentrations of Spirulina (25, 50, 75, and 100g in 5 litres of water) respectively. The zinc content of the yield of the cultivars was estimated and the study results indicated that there was a significant increase in zinc levels with p-value 0.015, 0.003 and 0.035 for Amaranthus gangeticus, Phaseolus aureus and Tomato respectively when compared with the control and between the set-ups, with biofortification of Spirulina platensis. The obtained results emphasize the application of Spirulina platensis to enhance the mineral nutrient in plants which are non-polluting, inexpensive, utilizable renewable resource to maintain the soil fertility.

Inf luence of Spirulina platensis exudates on the endocrine and nervous systems of a mammalian model

Objective: To investigate the ef ect of intra-peritoneal injection of purii ed exudates of axenic Spirulina platensis on the mammalian endocrine and nervous systems. Methods: The intra-peritoneal injection of the cyanobacterial exudates in mice was applied. Sex hormonal levels of testosterone and progesterone were measured using radioimmunoassay while the follicular stimulating hormone and luteinizing hormone were evaluated by direct chemiluminescence. In addition, superoxide dismutase, catalase and glutathione peroxidase were monitored in the hippocampus region using spectrophotometric method. The levels of the hippocampal monoamines, dopamine, noradrenaline and serotonin were analyzed by high performance liquid chromatography while the acetyl choline neurotransmitter was measured by colorimetric method using choline/acetylcholine assay kit. Results: A sharp disruption in the sex hormones levels of testosterone, progesterone, follicular stimulating hormone and luteinizing hormone was demonstrated in the serum of the treated mice. At the same time, a signii cant reduction in the endogenous antioxidant defense enzymes, superoxide dismutase, catalase and glutathione peroxidase was observed in the hippocampus region of the injected mice. Moreover, levels of dopamine, noradrenaline, serotonin and acetyl choline neurotransmitter in the same region were signii cantly af ected as a result of the treatment with Spirulina i ltrate. The gas chromatography-mass spectrometer and liquid chromatography mass spectrometry/mass spectrometry analysis showed the presence of some sterol-like compounds in the cyanobacterial i ltrate. Conclusions: This study demonstrated the capability of Spirulina to release detrimental bioactive metabolites into their surrounding that can disrupt the mammalian endocrine and nervous systems.

ISOLATION, PURIFICATION AND CHARACTERIZATION OF THE HYDROGEN EVOLUTION PROMOTING FACTOR OF HYDROGENASE OF SPIRULINA PLATENSIS

A component （s-factor） with obvious promoting effect on hydrogen evolution of hydrogenase has beenisolated and extracted from a Cell=free preparation of Spirulina platensis.The effect of the s-factor in the re-action system is similar to that of Na2S2O4, but is coupled with ligh. The s-factor has the maximumabsorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purifieds-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin,which has nochange of oxidized state and reduced state absorption spectra, and also has no promoting effect onhydrogenase of Spirulina platensis under the light.

EFFECT OF GERMANIUM DIOXIDE ON GROWTH OF SPIRULINA PLATENSIS

This study on the effect of different concentrations of germanium dioxide （GeO2） on the specificgrowth rate （SGR）, pigment contents, protein content and amino acid composition of Spirulina platensisshowed that Ge was not the essential element of this alga; that GeO2 could speed up growth and raiseprotein content of S. platensis, and could possibly influence the photosynthesis system. The concentrationrange of GeO2 beneficial to growth of S. platensis is from 5-100 mg/l. GeO2 is proposed to be util-ized to remove contamination by Chlorella spp. usually occurring in the cultivation of Spirulina.

Acclimatization of microalgae Arthrospira platensis for treatment of heavy metals in Yamuna River

Bioaccumulation and biosorption in microalgae are effective approaches for the removal of heavy metals(HMs)from river water.The objective of this study was to investigate the potential for use of acclimatized microalgae in the removal of HMs from the Yamuna River water as an acclimatizing medium.An active culture of Arthrospira platensis(A.platensis)was acclimatized to HMs up to a concentration of 100 mg/L.It was gradually exposed to increasing concentrations of HMs in five subsequent batches with a step increase of 20 mg/L to acclimatize live cells in the simulated Yamuna River water.The presence of high levels of HMs in the Yamuna River water caused growth inhibition.An empirical growth inhibition model was developed,and it predicted high threshold concentrations of HMs(210.7e424.5 mg/L),producing a positive specific growth rate of A.platensis.A.platensis also showed high average removal efficiencies of HMs,including 74.0%for Cu,77.0%for Cd,50.5%for Ni,76.0%for Cr,76.5%for Pb,and 63.5%for Co,from HMs-enriched Yamuna River water.The findings demonstrated that the maximum specific removal amounts of Cu,Cd,Ni,Cr,Pb,and Co were 54.0,58.0,39.0,62.8,58.9,and 45.3 mg/g,respectively.The maximum yields of the value-added products chlorophyll and phycocyanin were 2.5 mg/g(in a batch of 40 mg/L for Cd)and 1054 mg/g(in a batch of 20 mg/L for Cu),respectively.Therefore,acclimatized A.platensis was proven to be a potential microalga not only for sequestration of HMs but also for production of valuable pigments.

The Stabilization Effect of Glutaraldehyde on the Spirulina platensis Phycobilisomes

The spectral properties of the glutaraldehyde-treated phycobilisomes were studied. The results showed that glutaraldehyde was effective in preventing phycobilisomes from dilution- induced dissociation and preserving the intra-phycobilisomes energy transfer.

Isolation and Characterization of C-phycocyanin from Spirulina Platensis

The isolation of biliproteins from the Spirulina platensis cultured in southern China was accomplished with gel filtration on Sephacryl S 200 and chromatography on hydroxylapatite. The spectrophotometry, isoelectric point, and amino acid composition of C phycocyanin were determined, respectively.

THE EFFECT OF RADIOPROTECTION ON PLANT CELLS RADIOACTIVE DAMAGE BY POLYSACCHARIDE FROM SPIRULINA PLATENSIS

Comparative studies on vicia faba seeds were conductedbetwen treated group before radiation and treated oneafter radiation with polysaccharide from Spirulina platensis.The results of nuclear aberration frequency and chromosomalaberration types of vicia faba root_tip cells suggested