5th Day Of Culture Research Articles

Cell Weight, Chlorophyll a Content and Total Biomass Production of Spirulina platensis: Impact of Culture Media and Affiliated Correlation

This experiment was inquired to apprehend the associated relationship among the cell weight, chlorophyll a and total biomass production of Spirulina platensis grown in different concentration of digested rotten guava medium (DRGM) and kosaric medium (KM). This study was carried out to disclose the culture potentiality of different substrates and their impact on production through the assessment of their consequential correlation. The completely randomized design (CRD) was taken into consideration to accomplish the experiment in laboratory condition. The maximum cell weight was assumed in KM (12.41 ± 0.23) followed by 0.816 ± 0.0017, 0.813 ± 0.0013 and 0.807± 0.0011 mg/L in supernatant of 60%, 20% and 40% DRGM, respectively on the 10th day of culture. The correlation (P< 0.01) with the chlorophyll a content (r = 0.745) and total biomass (r = 0.741) revealed significant association and strong correlation with the obtained cell weight in these culture media. In kosaric medium, the growth performance was significantly higher (P<0.01) than that of grown in the different concentration of DRGM (60%, 40% and 20%). The content of chlorophyll a grown in KM (10.54 ± 0.33 mg/L) was significantly higher (P< 0.01) in comparison of 60% (0.863 ± 0.0018), 40% (0.769 ± 0.0015) and 20% DRGM (0.771 ± 0.13). The highest biomass of S. platensis was acquired in KM (705.52 ± 2.59 mg/L) and varied significantly (P < 0.05) than that of cultured in supernatants of 20% (67.76 ± 0.42), 40% (51.45 ± 0.26) and 60% (57.74 ± 0.26) DRGM. The culture techniques in DRG medium may dwindle the cost of production in comparison with KM and might be considered as a media for Spirulina platensis cultivation.

Stimulation of Lignan Production in Schisandra rubriflora In Vitro Cultures by Elicitation.

The study investigated the effect of elicitation with: chitosan (CH) (200 mg/L), yeast extract (YeE) (3000 mg/L), ethephon (ETH) (25 µM/L), and methyl jasmonate (MeJA) (50 µM/L), on lignan accumulation in agitated and bioreactor (Plantform temporary immersion systems) microshoot cultures of female (F) and male (M) Schisandra rubriflora Rehd. et Wils. (Schisandraceae) lines. The elicitors were supplemented on the 10th day of culture. Biomasses were collected at 24 h and 48 h, and 4, 6, and 8 days after the addition of each elicitor. The 24 compounds from the dibenzocyclooctadiene, aryltetralin, dibenzylbutane, and tetrahydrofuran lignans and neolignans were determined qualitatively and quantitatively in biomass extracts using the UHPLC–MS/MS method. The highest total contents [mg/100 g DW] of lignans were: for CH-95.00 (F, day 6) and 323.30 (M, 48 h); for YeE 104.30 (F, day 8) and 353.17 (M, day 4); for ETH 124.50 (F, 48 h) and 334.90 (M, day 4); and for MeJA 89.70 (F, 48 h) and 368.50 (M, 24 h). In the biomass extracts of M cultures grown in bioreactors, the highest total lignan content was obtained after MeJA elicitation (153.20 mg/100 g DW). The maximum total lignan contents in the biomass extracts from agitated and bioreactor cultures were 3.29 and 1.13 times higher, respectively, than in the extracts from the non-elicited cultures. The poor understanding of the chemical composition and the lack of studies in the field of plant biotechnology of S. rubriflora emphasize the innovativeness of the research.

Potentiality of Digested Rotten Guava Medium (DRGM) in Replacement of Kosaric Medium (KM): Perspective of Spirulina platensis culture

This study was conducted to evaluate the culture potentiality and growth performance of Spirulina (Spirulina platensis) in supernatant digested rotten guava (DRG) in replacement of Kosaric Medium (KM). The Spirulina was inoculated to grow in digested rotten guava media (DRGM) (treatments) with the addition of 9.0 g/L NaHCO3 and micronutrients and KM for a period of 14 days. The cell weight of spirulina was attained a maximum of 12.43±0.20 mg/L (dry wt. basis) in KM followed by 0.818 ± 0.003, 0.815 ± 0.0015 and 0.809± 0.0012 mg/L in supernatant of 60 (T1), 20 (T2) and 40% DRGM (T3), respectively on the 10th day of culture. The cell weight of spirulina grown in these media had highly significant (P<0.01) correlation with the chlorophyll a content (r = 0.746) and total biomass (r = 0.742) of Spirulina. The results showed that the growth performance of spirulina in supernatant of 60% DRGM was significantly (P<0.01) higher than that of spirulina grown in supernatant of 20% and 40% DRGM. The physico-chemical parameters viz. light intensity (2748 to 2768 lux/m2/s), temperature (19.0 to 22.2°C), pH (8.1 to 10.6), alkalinity (1522 to 2698 mg/L), nitrate-N (1.25 to 3.64 mg/L) and phosphate-P (11.30 to 55.40 mg/L) were within optimum level during the culture period. The results showed that, the different concentration of digested rotten guava (20%, 40%, 60%) has potential to increase the growth rate of Spirulina. Therefore, the DRG medium may be commercially used for mass culture of Spirulina platensis.

Proximate Composition Analysis of Spirulina platensis in Lab Scale Cultivation: Prospects of Digested Rotten Guava as a Culture Media

An experiment was conducted to evaluate the proximate composition of Spirulina platensis and growth performance in supernatant of different concentrations (supernatant of 20, 40 and 60 rotten guavas) of digested rotten guava medium (DRGM). The growth rates in terms of optical density, dry cell weight and chlorophyll a of spirulina were varied from each other. The cell weight of Spirulina was attained a maximum of 0.818 ± 0.003 mg/L in 60% DRGM followed by 0.815 ± 0.0015 and 0.809± 0.0012 mg/L in supernatant of 20 and 40%, respectively on the 10th day of culture. Similar trend was also observed in the case of optical density of the media contained Spirulina, chlorophyll a content (mg/L), total biomass (mg/L), specific growth rates (on the basis of cell weight and chlorophyll a) and total biomass of Spirulina. Cell weight of Spirulina grown in these media had highly significant (P < 0.01) correlation with the chlorophyll a content (r = 0.746) and total biomass (r = 0.742) of Spirulina. The growth performance of Spirulina in supernatant of 60% DRGM was significantly (P < 0.01) higher than that of grown in 20% and 40% DRGM. The percentage of crude protein was found 53.35 ± 0.32% in supernatant of 40% DRGM. The crude lipid was attained of 10.15 ± 0.14% in supernatant of 60% DRGM which was significantly (P < 0.05) higher than that of grown in 20% and 40% of DGRM.

Growth performance of Spirulina platensis in supernatant of digested rotten guava

This study was performed to evaluate the culture potentiality and growth performance of Spirulina (Spirulina platensis) in supernatant of three different concentrations of digested rotten guava (DRG) and Kosaric Medium (KM) as control. The study consisted of three different concentrations of rotten guavas, such as 20 (T1), 40 (T2) and 60 (T3) were allowed to digest under aeration. After 34 days, three replications showed the reddish white-colored supernatant screened and taken in a 2.0 L conical flask. Then, Spirulina was inoculated to grow in these three digested rotten guava media (DRGM) with the addition of micronutrients and KM and 9.0 g/L NaHCO3 for 14 days. The cell weight of Spirulina was attained at a maximum of 12.43±0.10 mg/L (dry wt. basis) in KM, followed by 0.818 ± 0.002, 0.815 ± 0.0014 and 0.809± 0.0011 mg/L in supernatant of 60, 20 and 40% DRGM, respectively on the 10th day of culture. A similar trend was also observed in the case of specific growth rates (based on cell weight and chlorophyll a) and total biomass (mg/L) of Spirulina. Cell weight of Spirulina had a highly significant (P<0.01) correlation with the chlorophyll a content (r= 0.746) and total biomass (r= 0.742) of Spirulina. The results on the growth performance showed that Spirulina grown in supernatant of 60% DRGM was significantly (P<0.01) higher (T3) than that of Spirulina grown in supernatant of 20% (T1) and 40% DRGM (T2).

CO2 sequestration using a novel Belt Conveyor Reactor with rotating sieve trays compared with Airlift Bubble Column as photobioreactors

The present work aims to evaluate the performance of a novel three-phase reactor Belt Conveyor Reactor BCR (with rotating sieve trays patent GB2567340B) as a photobioreactor compared with a traditional Airlift Bubble Column ALR, both influenced by the flow of the gas. Chlorella vulgaris was cultivated in these two photobioreactors using ambient air has a CO2 concentration of 0.038% with different aeration flowrates 0.145, 0.195, 0.24, and 0.29 vvm (gas volume per minute/liquid volume in the reactor).The maximum growth rate achieved on the 14th day of culture was 2.120 and 1.420 g/L for BCR, ALR respectively, with initial biomass concentrations of 0.2 g/L and aeration flow of 0.29 vvm. Moreover, the removal efficiency of carbon dioxide sequestration by the two photobioreactors is 40% for BCR and 25% for ALR. The innovative design succeeded in operational quality of agitation with high gas holdup inside the sieve trays to increase the biomass growth up to 50% higher than in the ALR. The maximum CO2 fixation for ALR was at about 18% at a specific aeration rate of 0.145 vvm in the bubbly regime. Whereas in the BCR can be reached about 38% with wide range operation condition of airflow rate 0.145–0.24 vvm due to well-distributed liquid due to rotating trays, and good gas–liquid mass transfer surface area.

Effects of nanometer magnesium hydroxide on soil cadmium form under different cadmium levels

AbstractCd is one of the most serious and harmful heavy metals in the world. Nanomaterials as a new passivating agent for heavy metals in soil have become a hot issue. But nanometer magnesium hydroxide application to heavy metal pollution in soils has not been reported. The laboratory culture experiments were conducted to study the effect of nanometer magnesium hydroxide and ordinary magnesium hydroxide (100, 200, and 300 mg kg−1) as cadmium passivators on soil cadmium form under different cadmium levels (1, 5, 10, and 15 mg kg−1). The results showed that the soil Cd distribution ratio of exchangeable Cd (EX‐Cd FDC) was 66.7% to 81.8% at cadmium levels of 1, 5, 10, and 15 mg kg−1, identifying it as the main form of soil cadmium. Soil EX‐Cd FDC reached the lowest level on the 14th day of culture. Addition of both nanometer magnesium hydroxide and ordinary magnesium hydroxide decreased the EX‐Cd FDC of the soil at each cadmium level. With increasing amount, the soil EX‐Cd FDC followed a decreasing trend. During the 0–28 days of culture, at the levels of 1, 5, 10, and 15 mg kg−1 cadmium, the EX‐Cd FDC of soil treated with nanometer magnesium hydroxide and ordinary magnesium hydroxide decreased by 11.4% to 67.7%, 7.8% to 37.2%, 7.7% to 36.4%, and 5.0% to 28.8% (nanometer magnesium hydroxide) and .5% to 49.5%, .6% to 15.0%, 1.0% to 18.1%, and .7% to 14.6% (common magnesium hydroxide), respectively, compared with control. Under the same application amount, nanometer magnesium hydroxide achieved a better effect than ordinary magnesium hydroxide in reducing soil EX‐Cd FDC. The antagonism between Cd and Mg competes for exchange sites during soil adsorption or Mg2+ forms hydroxy‐magnesium through OH− coordination in the soil via hydrolysis equilibrium. This facilitates the formation of precipitates, such as carbonates in Cd2+, and reduces active‐state cadmium in soil. The effect of nanometer magnesium hydroxide in passivating active cadmium on cadmium‐contaminated soil exceeds that of common magnesium hydroxide.

Multivariate analysis of the factors associated with live births during in vitro fertilisation in Southeast Asia: a cross-sectional study of 104,015 in vitro fertilisation records in Taiwan.

To investigate the factors associated with live births and the interaction between age and the number of embryos transferred after in vitro fertilisation (IVF) treatment. This study analyses data from a population-based-assisted reproductive database of all registered artificial reproduction institutions (n = 80) from 2010 to 2016 in Taiwan. The probability of a live birth in correlation with the treatment parameters was measured with multivariate logistic regression analyses using the generalised additive model (GAM) and Pearson's chi-square exact test. A total of 104,015 IVF treatments performed between 2010 and 2016 were included in our analysis. From these treatments, 31,467 (30.3%) were successfully delivered, and 40,565 test-tube babies were born. Pearson's chi-square exact test indicated that parents' age, cause of infertility, type of ovarian stimulation, additional assisted reproductive technology techniques, donated egg or sperm, fresh or frozen embryo, presence or absence of ovarian hyperstimulation syndrome, and day of embryo transfer were significantly associated with live births after an IVF cycle (p < 0.05). Multiple logistic regression analysis with the GAM revealed that the odds of a live birth with IVF treatment in patients < 34 years of age were 2.55 times higher than that in patients ≥ 45 years of age (odds ratio = 2.55, 95% confidence interval = 1.69-2.90) for patients who underwent a single-embryo transfer (SET); a similar pattern was observed when two or more embryos were transferred. Egg donation, the assisted hatching technique, oral ovarian stimulation agents, and implantation of frozen embryos during SET were shown to improve the chance of a live birth by 29-90%. Implantation of the embryo after the 5th day of culture yielded the highest odds of a live birth. The interaction plot revealed that maternal age, especially < 40 years, was associated with the probability of a live birth. SET and double-embryo transfer showed similar associations with the probability of a live birth across age groups. Transferring more than two embryos might reduce the probability of a live birth during IVF treatment for women ≥ 40 years of age. Implanting a greater number of embryos did not improve the age-related decrease in fertility for patients undergoing IVF. Therefore, we suggested that ≤ 2 blastocysts could be transferred during IVF treatments for women ≥ 40 years. Transferring a blastocyst on day 5 of culture was associated with a significant increase in the odds of a live birth resulting from IVF.

Smart Production of Lipids as Bio-Fuel in Spirulina Platensis (=Arthrospira Fusiformis), and Bio-Oxygen and Bio-Electricity in Media Cultured in Supernatant of Digested Poultry Waste

An experiment was conducted to evaluate growth performances and production of bio-fuel of Spirulina platensis (Gomont), and bio-oxygen and bio-electricity of culture media in supernatant of three different amount of digested poultry waste (DPW), and Kosaric medium (KM) as control. Three different amounts (concentrations) such as 2.0, 4.0 and 6.0 g/L poultry waste were allowed to digest under aeration. After 17 days, 700 ml grayish coloured supernatant was taken with addition of 9.0 g/L NaHCO3 and 0.50 ml/l micronutrient in 2.0 L conical flask with three replications and then autoclaved. Spirulina was inoculated to grow in these three treatments including KM (Control) after 72 hours of autoclave and then allowed to grow for a period of 14 days. This duration was estimated through repeated growth trials. The cell weight of spirulina was attained a maximum of 12.58 ± 1.25 mg/L (dry wt. basis) in KM followed by 11.46 ± 1.03, 9.16 ± 0.84 and 8.13 ± 0.73 mg/L in supernatant of 4.0, 2.0 and 6.0 g/L DPW, respectively on the 10th day of culture. Similar trend was also observed in the cases of optical density of the media contained spirulina, chlorophyll a content (mg/L), cell weight, total biomass (mg/l) and specific growth rates on the basis of cell weight and chlorophyll a. Cell weight of spirulina grown in these media had highly significant (P &lt; 0.01) correlation with the chlorophyll a content (r = 0.993) of spirulina. Crude protein of spirulina grown in supernatant of DPW was lower than that of spirulina cultured in KM. Crude lipids as bio-fuel of spirulina cultured in supernatant of 4.0 g/L DPW was almost three times higher than that of spirulina grown in KM which may be due to high phospholipids bioaccumulation. Phosphate-P was decreased in media of DPW due to its use for high lipids biosynthesis as phospholipids. Bio-oxygen was produced higher ranged from 2.1 ± 11 mg/L on initial day to 10.20 ± 0.54 mg/L on 10th day grown in supernatant of 4.0 g/L DPW media. Bio-electricity as green electricity ranged from 135 ± 4 on initial day to 284 ± 7 mV on 10th day when spirulina cultured in supernatant of 4.0 g/L DPW which was higher than grown in other media. pH followed the similar trend like bio-oxygen and bio-electricity. It was found that the production of bio-oxygen, bio-fuel and bio-electricity had direct relation with pH in culture media of spirulina. Therefore, mass production of Spirulina platensis might be done in supernatant of 4.0 g/L digested poultry waste to get high total lipids as bio-fuel, bio-electricity and bio-oxygen.

ISOLATION AND CHARACTERIZATION OF AMMONIA EXCRETING CYANOBACTERIUM CYLINDROSPERMUM SP. NDOP002

Cylindrospermum sp. NDOP002 was isolated from agricultural fields of Azamgarh, U.P., India. It was characterized by morphological methods. Total chlorophyll a content was 18.5 µg/ mg dry weight. Organism achieved a stationary phase of growth after 15 days of cultivation. Ammonia excretion was monitored from 6 hrs. of culture to 20 days. The ammonia content of culture increased up to 7 days (Maximum amount of 3.23 µg/ ml) and then decreased. Approximately a constant amount of ammonia was maintained from 9th and onwards days of growth. Maximum glutamine synthetase (GS) activity of 8.33 mM ?glutamyl hydroxamate min-1 mg chl-1 was observed on the 5th day of culture and then decreased. Approximately constant GS activity was observed after 9th and onward days of growth. Algalisation experiment showed an increase in growth parameter of rice plant in algalised set. Length of roots was 6.4-6.9 cm in non algalised and 7.2-7.6 cm in algalised rice plants whereas the length of shoots was 5.7-6.1 cm in non- algalised and 9.5-10.1 cm in algalised rice plants. The amount of ammonia in algalised set was 2.1 µg/ ml on the 10th day of the experiment. Experimental findings clearly suggest Cylindrospermum sp. NDOP002 as suitable inocula for algalisation of rice fields of Azamgarh district, U.P., India.

Smart Culture of Spirulina Using Supernatant of Digested Rotten Tomato (Solanum Lycopersicum) to Produce Protein, Bio-Fuel and Bio-Electricity

An experiment was conducted for the production of protein, bio-fuel and bio-electricity from the culture system of Spirulina platensis (Gomont) in supernatant of three different amount of digested rotten tomato (Solanum lycopersicum), and Kosaric Medium (KM) as control. Three different concentrations such as 25, 50 and 75% rotten tomato were allowed to digest under aeration. After 17 days, the colorless supernatant was screened and taken in 1.0 L conical flask with three replications. Then, Spirulina platensis was inoculated to grow in these three media (treatments) with the addition of 9.0 g/L NaHCO3 and micronutrients, and also in KM as control for a period of 14 days. The cell weight, optical density, chlorophyll a and total biomass of spirulina was attained to the maximum values when grew in KM on the 10th day of culture followed by supernatant of 50% digested rotten tomato (DRT) than in 25 and 75% DRT culture. The chemical properties of the culture media such as pH, salinity, dissolved bio-oxygen, electric conductivity and bio-electricity were increased from first day up to 12th day of experiment. Total biomass of spirulina grown in these media had highly significant (P &lt; 0.01) correlation with cell weight (r = 0.825) and chlorophyll a (r = 0.866) of spirulina. The results showed that the growth performances of S. platensis grown in supernatant of 50% DRT was significantly (P &lt; 0.01) higher than that of spirulina grown in supernatant of 25 and 75% DRT. The percentage of crude protein (55.10 ± 0.45 to 59.90 ± 0.33%) of spirulina grown in supernatant of DRT was little bit higher than that of spirulina cultured in KM (58.40 ± 0.38%). But bio-fuel in terms of crude lipids (16.50 ± 0.31%) of spirulina cultured in supernatant of 50% DRT was almost two and half times higher than that of spirulina grown in KM (crude lipids, 6.30 ± 0.21%). Bio-electricity (300 ±10.20 mV) produced in culture of spirulina in supernatant of 50% DRT was higher than that recorded in KM (240 ±10.20 mV) followed by 75% DRT and other media. Bio-electricity had directly and strongly significant (p &lt; 0.001) correlation with pH (r = 0.812), dissolved bio-oxygen (r = 0.832), salinity (r = 0.788) and electric conductivity (r = 0.856). Therefore, this procedure will produce huge amount of electricity in the world and will make a revolution in this field of bio-electricity production. Whole world will be benefited from the output (results) of this experiment.

Smart production of spirulina (Spirulinaplatensis) using supernatant of digested rotten potato (Solanum tuberosum)

The study was conducted to evaluate the culture and growth performance of spirulina (Spirulina platensis) in supernatant of three different amounts of digested rotten potato (DRP), and Kosaric medium (KM) as control in 16 days after 26 days digestion. Three different concentrations such as 20, 40 and 60% of DRP were used. Spirulina was inoculated in supernatant DRP along with 9.0 g/L NaHCO3 and micronutrients, and KM for a period of 14 days. The cell weight of spirulina was attained a maximum of 12.42 ± 0.21 mg/L in KM followed by 8.352 ± 0.21, 6.256 ± 2.34 and 9.505 ± 0.43 mg/L in supernatant of 40, 20 and 60% DRP, respectively on the 10th day of culture. Similar trend was also observed in the cases of optical density, chlorophyll a, total biomass, specific growth rates. Cell weight of spirulina grown in the media were highly significant (p<0.01) and correlated with the chlorophyll a content and total biomass. The growth performance of spirulina grown in supernatant of 60% DRP was significantly higher than that of spirulina grown in supernatant of 20and 40% DRP. Therefore, mass culture of spirulina may be done in supernatant of 60% DRP.

Toxicity removal and biodegradability enhancement of sludge extract in hydroquinone-rich wastewater via cultivation of Chlorella vulgaris

Excess sludge is rich in organic matter, nitrogen, and phosphorus. However, in the disposal of industrial wastewater with high concentrations of organic matter, the toxic organics accumulate in the sludge will cause sludge toxicity. Microalgae have strong adaptability to toxic environments. Therefore, in the present study, Chlorella vulgaris was cultured in sludge extract and BG11 medium (blank group) with the objectives of reducing both sludge yield and organic toxicity. The results show that the toxic sludge extract could maintain the stable growth of Chlorella vulgaris, and on the 10th day of culture, Chlorella vulgaris obtained higher biomass and photosynthesis efficiency. The BOD/COD (B/C) ratio of the sludge extract increased from 0.12 to 0.37 while the carbon/nitrogen ratio increased by a factor of 4.9, which significantly improved its biodegradability. Excitation emission matrix analysis showed that C. vulgaris removed soluble microbial products and reduced some humic acids. The isobaric tags for relative and absolute quantitation revealed the presence of differential proteins affecting the processes of small molecule metabolism, energy metabolism, and photosynthesis in different media.

Early Addition of Parathyroid Hormone-Related Peptide Regulates the Hypertrophic Differentiation of Mesenchymal Stem Cells.

Chondrogenic differentiation of mesenchymal stem cells (MSCs) into hyaline cartilage is complicated by terminal hypertrophic differentiation. In growth plate, parathyroid hormone-related peptide (1-34) (PTHrP) plays a crucial role in maintaining chondrocytes in their proliferation state by counteracting the hypertrophic differentiation. This study aims to test the effect of PTHrP supplementation at different time points on chondrogenic differentiation of MSCs and assess the final quality of differentiated chondrocytes. Human periosteum and bone marrow MSCs isolated from 3 patient samples (donor unmatched) were characterized by flow cytometry and multilineage differentiation. The cells were differentiated into chondrocytes in the presence of transforming growth factor-β (TGF-β) and the PTHrP (1-34) was added from 4th or 14th day of culture. The outcome was analyzed by histology, immunohistochemistry, and gene expression. Flow cytometry and multilineage differentiation confirmed that the cells isolated from periosteum and bone marrow exhibited the phenotype of MSCs. During chondrogenic differentiation, pellets that received PTHrP from the 4th day of culture showed a significant reduction in hypertrophic markers (COL10A1 and RUNX) than the addition of PTHrP from the 14th day and TGF-β alone treated samples. Furthermore, 4th day supplementation of PTHrP significantly improved the expression of cartilage-specific markers (COL2A1, SOX9, ACAN) in both periosteum and bone marrow-derived MSCs. Histology and immunostaining with collagen type X data corroborated the gene expression outcomes. The outcome showed that supplementing PTHrP from the 4th day of chondrogenic differentiation produced better chondrocytes with less hypertrophic markers in both bone marrow and periosteal-derived MSCs.

Study on the Active Effect of Soil Nutrient Elements

The purpose of this study is to ecologically soil, reduce fertilization and maintain land productivity. In this study, the amount of nutrient elements stored in the soil is large, and the soil samples are treated with long-term constant and different water content to determine the activation amount of the solid elements in the soil. The results showed that the content of available phosphorus and available potassium released by XH6 in different water content cultures was higher than 3.89% and 10.12% of XH10, respectively. The content of nitrate nitrogen released by XH4 was higher than that of treated XH4. %. The content of ammonium released under treatment of XH10 was higher than that of treatment with XH10 of 24.11%. The release nutrients varied with the culture time, but the release of nutrients such as ammonium nitrogen, nitrate nitrogen, available phosphorus and available potassium peaked on the 14th day of culture, and stabilized in the latter culture. So, the high moisture content has a certain release and increase effect on soil available potassium and available phosphorus.

Silicate-substituted strontium apatite nano coating improves osteogenesis around artificial ligament

BackgroundTreatment of anterior cruciate ligament injuries commonly involves the use of polyethylene terephthalate (PET) artificial ligaments for reconstruction. However, the currently available methods require long fixation periods, thereby necessitating the development of alternative methods to accelerate the healing process between tendons and bones. Thus, we developed and evaluated a novel technique that utilizes silicate-substituted strontium (SrSiP).MethodsPET films, nano-coated with SrSiP, were prepared. Bone marrow mesenchymal cells (BMSCs) from femurs of male rats were cultured and seeded at a density of 1.0 × 104/cm2 onto the SrSiP-coated and non-coated PET film, and subsequently placed in an osteogenic medium. The osteocalcin concentration secreted into the medium was compared in each case. Next, PET artificial ligament, nano-coated with SrSiP, were prepared. BMSCs were seeded at a density of 4.5 × 105/cm2 onto the SrSiP-coated, and non-coated artificial ligament, and then placed in osteogenic medium. The osteocalcin and calcium concentrations in the culture medium were measured on the 8th, 10th, 12th, and 14th day of culture. Furthermore, mRNA expression of osteocalcin, alkaline phosphatase (ALP), bone morphogenetic protein-2 (BMP2), and runt-related transcription factor 2 (Runx2) was evaluated by qPCR. We transplanted the SrSiP-coated and non-coated artificial ligament to the tibiae of mature New Zealand white rabbits. Two months later, we sacrificed them and histologically evaluated them.ResultsThe secretory osteocalcin concentration in the medium on the film was significantly higher for the SrSiP group than for the non-coated group. Secretory osteocalcin concentration in the medium on the artificial ligament was also significantly higher in the SrSiP group than in the non-coated group on the 14th day. Calcium concentration on the artificial ligament was significantly lower in the SrSiP group than in the non-coated group on the 8th, 10th, 12th, and 14th day. In qPCR as well, OC, ALP, BMP2, and Runx2 mRNA expression were significantly higher in the SrSiP group than in the non-coated group. Newly formed bone was histologically found around the artificial ligament in the SrSiP group.ConclusionsOur findings demonstrate that artificial ligaments using SrSiP display high osteogenic potential and thus may be efficiently used in future clinical applications.

Bovine pericardium membrane as new tool for mesenchymal stem cells commitment

Acellular matrices are widespread biomaterials used in surgical practice as tissue reinforcement and anatomical support to favor tissue regeneration. It is clear that a fundamental role in the regeneration of tissue is played by cell-material interaction. In this work, the interaction between a bovine pericardium membrane and human adult stem cells was investigated by microscopy analysis and gene expression analysis. Parallel cell cultures were prepared on the pericardium membrane or tissue culture plate. They were incubated in basal growth medium or in adipogenic differentiation medium to perform experiments on the seventh and the 14th day of culture. Results demonstrated that the membrane allows cell viability, adhesion, and proliferation of human stem cells. During adipogenic commitment on the membrane, the accumulation of cytoplasmatic lipid droplets and the expression of adipogenic gene PPARG, CEBPA, GLUT4, FABP4, and ADIPOQ were detected. Concurrently, a downregulation of mesenchymal stem cell gene CD29, CD90, and CD105 was detected. In basal medium, the adipogenic gene expression was upregulated, whereas the mesenchymal markers were indifferently expressed. These findings suggest that the bovine pericardium membrane is a biocompatible matrix and that their rough surface allows cell adhesion, spreading, and proliferation. The surface morphology activates mechanochemical signals that stimulate the adipogenic commitment of stem cells in basal medium and potentiate their commitment in adipogenic differentiation medium.

Evaluation of rotten orange as a source of inorganic nutrients for blue green algae, Spirulina platensis culture

A study was conducted to examine the culture and growth performance of Spirulina platensis in three different concentrations (25, 50 and 75 percent) of digested rotten orange (DRO) and Kosaric medium (KM) as control for three months. Each experiment was done in triplicates under fluorescent light in light: dark (12 hour: 12 hour) condition for a period of 14 days. Cell weight, chlorophyll a content of Spirulina platensis and physico-chemical parameters of cultured media (temperature, pH, light intensity) were measured at every alternative day. During the culture period, temperature was more or less similar and ranged from 30.1 to 31.4˚C. The range of light intensities was recorded from 2660 to 2770 lux/ /s during the culture period. The maximum pH was recorded 9.9 in 25% digest rotten orange (DRO) on 8th day of the culture and minimum was recorded 8.2 in all the medias on the initial day of the culture. The growth rate of Spirulina platensis cultured in supernatant of DRO and KM were varied. The initial cell weight of Spirulina platensis was 0.0023 mg/L that attained maximum cell weight of 12.44 mg/L in Kosaric medium on the 10th day of culture. Besides that, the maximum cell weight was 7.679, 12.366, and 9.455 mg/L when cultured in supernatant of 25, 50, and 75% in DRO on the 10th day of the culture. While, the initial chlorophyll a content of Spirulina platensis was 0.0015 mg/L which attained the highest content of 10.54 mg/L when grown in KM. The highest chlorophyll a content of Spirulina platensis was 6.926 mg/L when grown in 25% DRO, it was 10.476 mg/L when grown in 50% DRO, and it was 9.140 mg/L when grown in 75% DRO on the 10th day of the culture. This study showed that, the growth performance of Spirulina platensis was higher in supernatant of 50% DRO than 25 and 75% DRO. This concentration of medium gave satisfactory results compared with standard KM. This variation probably occurred because of differences in nutrient concentrations and composition of the medium. Therefore, it can be concluded that, the concentration of 50% DRO is suitable and favorable for Spirulina platensis culture.&#x0D; Asian Australas. J. Biosci. Biotechnol. 2018, 3 (3), 208-224

Establishment of an improved vitrification protocol by combinations of vitrification medium for isolated mouse ovarian follicles

In vitro follicle growth (IVFG) is an emerging alternative option for fertility preservation in women instead of ovarian tissue cryopreservation and transplantation. To widen the application of this technique, follicle cryopreservation should be established prior to clinical use. In the present study, we tried to determine the optimal vitrification protocol of mouse ovarian follicle for in vitro culture and oocyte maturation by comparing four different compositions of cryoprotective agents (CPA). Secondary follicles were mechanically isolated from 2-week-old BDF-1 mice and randomly assigned to fresh control and four different groups by the composition of CPAs (ES, EDS, EFS and EPS groups; E: ethylene glycol, D: dimethyl sulfoxide, S: sucrose, F: ficoll, P: 1,2-propanediol (PROH)). After vitrification and warming procedures, the follicles were cultured in vitro for 10 days and then treated with human chorionic gonadotropin and epidermal growth factor to induce oocyte maturation. Fourteen to 16 h later, oocyte maturation and quality were assessed. Follicle viability was evaluated by Calcein-AM/ethidium homodimer-1 staining immediately after warming, and their survival and diameters were measured during follicle culture periods. Antral cavity formation was observed at the end of the culture period (on the 10th day of culture). Following oocyte maturation, its maturational ability and meiotic spindle formation were assessed to evaluate their competence. There was no significant difference in viability after warming among the vitrification groups. From the 8th day of culture, the survival rate of ES and EDS were significantly higher than those of other vitrification groups (EPS and EFS). The follicle diameter was largest in the fresh-control group from the 6th day, while smallest in the EFS with statistical significance. On the 10th day of culture, the antral-cavity formation rate of EDS was comparable to that of the fresh control group. However, the oocyte maturation was significantly decreased in all four vitrification groups when compared with control group; especially, the EFS showed a more marked reduction in the oocyte maturation. There were no significant differences in meiotic spindle formation among all of those groups. Our results suggest that EDS combination for mouse follicle vitrification are the most effective vitrification protocols for mouse follicle and evaluated by an in vitro culture and oocyte maturation after warming.

Sequential medium with GH and IGF-1 improved in vitro development of bovine preantral follicles enclosed in ovarian tissue.

The growth hormone (GH) and growth insulin-like factor-1 (IGF-1) act directly upon the regulation and growth in the different phases of preantral follicles. Thus, it is necessary to define their sequentiality until the in vitro preovulatory development. Therefore, the study aimed to assess the effects of a sequential medium containing GH and/or IGF-1 in the long-duration in vitro culture of preantral ovarian follicles. Ovarian fragments were cultivated: first half (days 1-7), second half (days 7-14) or during 14 culture days. Treatments were identified as: αMEM+; GH→IGF-1; IGF-1→GH and GH+IGF-1. The culture was designed in 24-well plates, in an incubator at 37°C and 5% CO2 . The parameters of normality, viability, follicles (primordial/in developing) and follicle diameter were evaluated. In addition, the ultrastructure was confirmed with electron transmission microscopy. The results showed that the culture treated with GH→IGF-1 kept the follicular normality and the viability until the 14th day of culture and increased both in the follicular development until 7th day and in the follicular diameter until 14th day, when compared to the control. The treatments IGF-1→GH and GH+IGF-1 were not effective in the developing and follicular diameter after 7days of culture, and also reduced the percentage of viability. It is concluded that the bovine preantral follicles cultured in the sequential medium treated with GH→IGF-1 improved the follicular development until the first half of the culture and kept these parameters with normality, viability and ultrastructure until the second half of the in vitro culture.