Patterns in the composition of the rotifer communities from high mountain lakes and ponds in Sierra Nevada (Spain)

On the basis of periodic collections of rotifers from 29 lakes and ponds over 2500 m above sea level in the Sierra Nevada (Southern Spain), patterns of species richness, distribution and community composition were evaluated. Results allow us to distinguish communities which fall into two major lake types. One is defined by the presence of typically planktonic species as well as lower specific richness whereas the other includes communities of mainly benthic and periphytic species. Both lake types seem to be related to small differences in their physical and chemical characteristics. These relationships and the influence of littoral vegetation are discussed.Research supported by CAICYT Project n° 3069/83     

Endothelial lipase releases saturated and unsaturated fatty acids of high density lipoprotein phosphatidylcholine

We assessed the ability of endothelial lipase (EL) to hydrolyze the sn-1 and sn-2 fatty acids (FAs) from HDL phosphatidylcholine. For this purpose, reconstituted discoidal HDLs (rHDLs) that contained free cholesterol, apolipoprotein A-I, and either 1-palmitoyl-2-oleoylphosphatidylcholine, 1-palmitoyl-2-linoleoylphosphatidylcholine, or 1-palmitoyl-2-arachidonylphosphatidylcholine were incubated with EL- and control (LacZ)-conditioned media. Gas chromatography analysis of the reaction mixtures revealed that both the sn-1 (16:0) and sn-2 (18:1, 18:2, and 20:4) FAs were liberated by EL. The higher rate of sn-1 FA cleavage compared with sn-2 FA release generated corresponding sn-2 acyl lyso-species as determined by MS analysis. EL failed to release sn-2 FA from rHDLs containing 1-O-1'-hexadecenyl-2-arachidonoylphosphatidylcholine, whose sn-1 position contained a nonhydrolyzable alkyl ether linkage. The lack of phospholipase A(2) activity of EL and its ability to liberate [(14)C]FA from [(14)C]lysophosphatidylcholine (lyso-PC) led us to conclude that EL-mediated deacylation of phosphatidylcholine (PC) is initiated at the sn-1 position, followed by the release of the remaining FA from the lyso-PC intermediate. Thin-layer chromatography analysis of cellular lipids obtained from EL-overexpressing cells revealed a pronounced accumulation of [(14)C]phospholipid and [(14)C]triglyceride upon incubation with 1-palmitoyl-2-[1-(14)C]linoleoyl-PC-labeled HDL(3), indicating the ability of EL to supply cells with unsaturated FAs.     

High density culture of the freshwater rotifer,Brachionus calyciflorus

The freshwater rotifer, Brachionus calyciflorus is one of the live food organisms used for the mass production of larval fish. In this study possibility of obtaining high density cultures of the freshwater rotifer B. calyciflorus were investigated. The two culture systems used differed in their air and dissolved oxygen supplies using three temperatures in each case: 24, 28 and 32 °C. Rotifers were batch-cultured using 5 l-vessels and fed with the freshwater Chlorella. The growth rate of rotifers significantly increased with an increase in temperature. The maximum density of the rotifers with air-supply at 24 °C, 6500 ind. ml^–1, was significantly lower than those cultured at 28 and 32 °C, i.e. 8600 and 8100 ind. ml^–1, respectively. Dissolved oxygen levels decreased with time and ranged from 0.8 to 1.4 mg l^–1 when the density of freshwater rotifer was the highest at each temperature. The highest density (19200 ind. ml^–1) of freshwater rotifer was obtained in cultures with a supply of oxygen at 28 °C. Densities of 13500 and 17200 ind. ml^–1 were found at 24 and 32 °C, respectively. Levels of NH₃-N increased with time and a dramatic increase of NH₃-N was observed at high temperatures. Levels of NH₃-N at 24, 28 and 32 °C were 13.2, 18.5 and 24.5 mg l^–1, respectively. These levels coincided with the highest rotifer density at each of the three temperatures. When rotifers were cultured with an oxygen-supply and pH was adjusted to 7, the maximum density of rotifer reached 33500 ind. ml^–1 at 32 °C . These results suggested that high density culture of freshwater rotifer, B. calyciflorus could be achieved under optimal conditions with DO value of exceeding 5 mg l^–1 and NH₃-N values of lower than 12.0 mg l^–1.     

Production of human-mouse chimeric antibody by high cell density perfusion culture

Two mouse myeloma cell lines which were transfected with chimeric mouse variable-human constant immunoglobulin heavy and light chain genes have been cultured at high cell density in a settling perfusion culture vessel to produce chimeric antibody specific for human common acute lymphocytic leukemia antigen (cALLA).J558L transfectant proliferated well in a serum-free medium (ITES-eRDF) to a viable cell density of 3.7×10⁷ cells/ml and produced chimeric antibody to a maximum value of 60 g/ml in 120 ml scale vessel. X63Ag8.653 transfectant reached a density of 1.9×10⁷ cells/ml in 1.2 I scale vessel in serum supplemented medium (10% FCS-eRDF) and produced chimeric antibody which consisted of chimeric gamma and chimeric kappa chains to a maximum value of 5.8 g/ml.     

Analysis of the growth performance,stress, profile of fatty acids and amino acids and cortisol in Tilapia (Oreochromis niloticus), cultured at high stocking density using in-pond raceway system

In-pond raceway system technology (IPRS) was introduced in Pakistan in 2019 as solution for sustainable aquaculture approach by effectively increasing production, reducing pollution and facilitating feed and pond management. Fingerlings of GIFT Tilapia (Oreochromis niloticus) (n = 16,500 in each raceway, initial weight = 32.00 ± 1.26 g) were stocked in June 2019 in two IPRS raceways (area of each raceway = 220 m³) for 171 days until harvested on November 30, 2019. Fingerlings stocked in traditional earthen ponds (area of each pond = 6167 m³) were studied as control (n = 3000 in each pond, initial weight = 32.00 ± 1.26 g). Average harvested biomass from raceways was 57.33 kg/m³ with an average daily weight gain per fish of 4.47 g per day. On the other hand, average harvested biomass from control ponds was 0.38 kg/m³ with an average daily weight gain per fish of 4.60 g per day. Average feed conversion ratio (FCR) in both raceways and control ponds was recorded as 1.25 and 1.24, respectively. Overall survival rate in both raceways and control ponds was above 99%. No sign of any disease was noted at any stage in both study groups. Crude protein and fats contents did not reduce in any raceway despite of high stocking density and sharp seasonal changes. Profile of essential and non-essential amino acids were found to be upto nutritional requirements of adult human. High levels of n-3 and n-6 fatty acids in fish collected from raceways as compared to those in traditional earthen pond show that muscle quality was not compromised due to high stocking density in small area. Return on investment excluding capital cost was 47.05 which implies that IPRS technology can be economically feasible with further modifications.     

酿酒酵母S.cerevisiae高密度培养条件优化研究

考察了培养基组成和培养条件对酿酒酵母Saccharomyces cerevisiae发酵的影响。以TB培养基为初始培养基,通过正交实验设计优化培养基组成,确定了影响酵母细胞产量最主要的因素是葡萄糖,最适培养基组成为:酪蛋白胨15 g/L,酵母粉25 g/L,葡萄糖30 g/L,KH2PO42.4g/L,K2HPO4.3H2O 16.34 g/L。并确定了最佳培养条件:温度30℃,转速150 r/min。采用优化培养基及培养条件下进行发酵,菌液最高OD600值和细胞密度分别达15.82和2.03×108/mL,比优化前分别提高24.2%和22.0%。     

Trophic transfer and trophic modification of fatty acids in high Arctic lakes

1. A comparative study of fatty acid (FA) profiles in particulate matter (seston) and the key grazer Daphnia was performed in six high Arctic ponds (79°N, Svalbard). The ponds were all small and shallow, but followed a strong gradient with respect to nutrient content and optical properties. 2. A distinct locality‐specific pattern was detected by principal component analysis of FA profiles, where samples from each locality clustered both with regard to seston and Daphnia. Linear discriminant analysis using nine sestonic fatty acids as discriminant functions gave on average a correct prediction of the Daphnia lake membership in 47% of cases, with very high predictability in some lakes but poor predictability in others. 3. No significant correlation was detected between FA and nutrient concentration or levels of dissolved organic carbon. The major determinant of FA profiles as judged from a redundancy analysis was the taxonomic composition of phytoplankton communities, notably the biomass of Chlorophyceae. 4. The FA profiles of Daphnia were for some FAs strongly enriched relative to the seston, while diluted for others. Among the polyunsaturated fatty acids (PUFAs), a pronounced magnification of eicosapentaenoic acid (EPA, 20 : 5 n‐3), and to some extent 18 : 3 n‐3 and 20 : 4 n‐6 was found, while docosahexaenoic acid (DHA, 22 : 6 n‐3) contributed in general less to FAs in Daphnia than in seston and was hardly detectable in Daphnia from most localities. 5. The overall content of PUFAs in Daphnia was consistently high, close to 40% of total FA in all investigated localities, despite major differences in seston PUFA content. Daphnia thus acts as a regulator with regard to overall PUFAs, reflecting its physiological constraints and relatively fixed demands for PUFAs in general. The distinct locality‐specific profiles in Daphnia strongly suggest a kind of FA‐fingerprint, but our data do not allow strict statements on the use of specific FAs as trophic markers.     

基因工程菌高密度发酵液中碳源物质甘油的定量检测及其浓度的优化控制

比较了高碘酸氧化法,铜离子络合法、高压液相色谱法和甘油激酶法四种不同的方法定量测定重组工程菌ＹＫ５３７／ＰＳＢ－ＴＫ高密度发酵液中甘油的浓度,发现甘油激酶法可以排除发酵液中其他物质的干扰,精确测定甘油的含量。在此基础上对发酵培养中甘油的浓度进行了优化,结果表明,甘油浓度控制在较低的水平有利于菌体密度的提高。在５Ｌ自动发酵罐中,控制甘油浓度在５ｇ／Ｌ左右,经２０ｈ的培养,最终细菌密度达到１２０ＯＤ６     

High density cell culture of a marine photosynthetic bacterium Rhodovulum sp. with self-flocculated cells

High density cell cultivation of a marine photosynthetic bacterium, Rhodovulum sp. PS88 with self-flocculated cells was established by using a single-tower fermenter. High density cell culture with continuous cultivation was yielded 43 g dry matter l–1 with acetate as a substrate consumed at 22.5 g/l day. © Rapid Science Ltd. 1998     

Substrates that limit high density cultures of Spodoptera frugiperda cells

In high density cultivation of Spodoptera frugiperda (Sf9) cells in Grace's medium supplemented with FBS (fetal bovine serum) and yeastolate, amino acids were the primary limiting substrates while the carbon sources were not. Glutamine, methionine, and threonine were consumed rapidly during the cultivation. When cultures were supplemented with amino acids, yeastolate components other than amino acids became the secondary limiting substrates.     

Formation of high density lipoproteins containing both apolipoprotein A-I and A-II in the rabbit

Human plasma HDLs are classified on the basis of apolipoprotein composition into those that contain apolipoprotein A-I (apoA-I) without apoA-II [(A-I)HDL] and those containing apoA-I and apoA-II [(A-I/A-II)HDL]. ApoA-I enters the plasma as a component of discoidal particles, which are remodeled into spherical (A-I)HDL by LCAT. ApoA-II is secreted into the plasma either in the lipid-free form or as a component of discoidal high density lipoproteins containing apoA-II without apoA-I [(A-II)HDL]. As discoidal (A-II)HDL are poor substrates for LCAT, they are not converted into spherical (A-II)HDL. This study investigates the fate of apoA-II when it enters the plasma. Lipid-free apoA-II and apoA-II-containing discoidal reconstituted HDL [(A-II)rHDL] were injected intravenously into New Zealand White rabbits, a species that is deficient in apoA-II. In both cases, the apoA-II was rapidly and quantitatively incorporated into spherical (A-I)HDL to form spherical (A-I/A-II)HDL. These particles were comparable in size and composition to the (A-I/A-II)HDL in human plasma. Injection of lipid-free apoA-II and discoidal (A-II)rHDL was also accompanied by triglyceride enrichment of the endogenous (A-I)HDL and VLDL as well as the newly formed (A-I/A-II)HDL. We conclude that, irrespective of the form in which apoA-II enters the plasma, it is rapidly incorporated into spherical HDLs that also contain apoA-I to form (A-I/A-II)HDL.     

Studies of the modulation of essential fatty acid metabolism by fatty acids in cultured neuroblastoma and glioma cells

In cultured neuroblastoma cells (N1E-115), the metabolism of the essential fatty acid, linoleic acid (18:2 (n-6)), to arachidonic acid (20:4(n-6)) can be altered by other fatty acids in a manner supporting a concerted action of the modulating fatty acid on the desaturation and chain elongation enzymes. In further examination of mechanisms involved, cultured glioma (C-6) or neuroblastoma-glioma hybrids (NG-108-15) cells showed similar patterns of activation by some fatty acids (e.g., 20:3(n-6) and 20:4(n-6)), and inhibition (e.g., 18:3(n-3) or 22:6(n-3)) or no effect (e.g., 18:1(n-9), 20:3(n-3)) by others. In contrast, only inhibition by 20:4(n-6) was seen in cultured HeLa cells, suggesting that the intracellular interactions may not be universal in all cell lines. For fatty acids that activate 20:4(n-6) formation, the lag observed when substrate and activator were administered simultaneously was eliminated by preincubation with activator. Maximal activation occurred within 4 h for neuroblastoma and 2 h for glioma; in each cell line activation declined steadily for 10 h after removal of the activator. Inhibition of protein synthesis did not alter activation. As 98% of the fatty acid incorporated was esterified to triacylglycerol or phospholipid and only the triacylglycerol mass expanded, several manipulations to potentially alter the flow of acyl chains between these lipid pools were evaluated using dual-label and pulse-chase experiments. Results suggested that competition between 18:2(n-6) utilization for esterification to phospholipid and the desaturation-chain elongation sequence as well as a more direct and specific interaction of certain fatty acids with the enzymes may influence 20:4(n-6) formation. A model to explain these observations is discussed.     

Polyunsaturated fatty acids in stream food webs – high dissimilarity among producers and consumers

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Differentiation and mineralization in chick chondrocytes maintained in a high cell density culture: A model for endochondral ossification

Summary Chondrocytes isolated from the proliferative and differentiating zones of 3-wk-old chick growth plates were cultured in the presence of 10% fetal bovine serum (FBS) and ascorbic acid for up to 21 d in a high cell density culture within Eppendorf tubes. The proliferative, differentiating, and calcification properties of the chondrocytes were examined by immunolocalization and by enzyme histochemical and biochemical methods. The cells maintained a chondrocyte phenotype throughout culture: they were round in shape and synthesized both collagen type II and proteoglycans. The expression of a hypertrophic phenotype was evident by Day 3 of culture and from this time onwards characteristics of terminal differentiation were observed. The cells were positive for both alkaline phosphatase (ALP) activity and c-myc protein and the surrounding matrix stained strongly for collagen type X. Small foci of mineralization associated with individual chondrocytes were first evident by Day 6 and more widespread areas of mineralization occupying large areas of matrix were present by Day 15. Mineralization occurred without the addition of exogenous phosphate to the medium. This culture system displays characteristics that are similar in both morphological and developmental terms to that of chick chondrocyte differentiation and calcification in vivo and therefore offers an excellent in vitro model for endochondral ossification.     

Nutrient optimization for high density biological production applications

Conclusion At the 1989 annual meeting of the U.S. Tissue Culture Associations, Ricahrd am, a leading investigator in the serum-free nutrient requirements of cultured cells, commented on the process of medium development. He noted that a survey of major media manufacturers revealed that, among the top selling mammalian cell culture media formulations, most were nearly thirty years old.This commentary is noteworthy considering the tremendous changes in cell culture understanding and derived applications which have emerged over these three decades. Fastidious cell types relatively unknown to investigators of the 1950s and 1960s are now being cultivated in defined, serum-free environments. Culture environments range from limiting dilution clonal recoveries to maintenance cultures approaching tissue densities. While research applications continue to predominate, applications of cell culture have expanded to the engineered production of biopharmaceuticals, to replacement of animal models for toxicology testing, and to the preservation, activation and expansion of human cells, tissues and organs.It is likely that future nutrient medium development will be predicated upon the design of a minimal number of defined formulations of relatively generic utility to a broad class of cell types. Analytical techniques derived from those described herein will be exploited in the user laboratory and in collaboration with the supplier to optimize the nutrient composition for the desired biological response.     

Very low density lipoproteins and lipoprotein lipase in serum of rats deficient in essential fatty acids

Rats fed a diet deficient in essential fatty acids have a low level of serum very low density lipoproteins (VLDL). It was found that after intraperitoneal injection of heparin, deficient rats had a higher level of lipoprotein lipase activity in their plasma than did normal rats. VLDL isolated from serum of normal and deficient rats were compared as substrates for postheparin lipase of rat plasma. There was no significant difference in V(max) between the two preparations of lipoproteins, but the apparent K(m) for lipoproteins from deficient animals was significantly less than that for normal animals. These observations suggest that the low concentration of VLDL in deficient rats may be explained (a) by an increased activity of lipoprotein lipase in the tissues of these animals and (b) by the VLDL of deficient rats being more rapidly hydrolyzed at low concentrations by lipoprotein lipase than VLDL from normal rats.     

基因工程菌高密度发酵工艺研究进展

阐述了基因工程菌高密度发酵工艺的几个主要影响因素,包括重组菌构建、培养条件、生长抑制因子以及它们的控制技术。通过高密度发酵可以提高细胞生长密度、目的蛋白的表达含量。在高密度发酵过程中,会产生一些有害抑制代谢副产物,但通过分批补料可以降低影响。     

Production of Chlorella vulgaris as a source of essential fatty acids in a tubular photobioreactor continuously fed with air enriched with CO2 at different concentrations

To reduce CO₂ emissions and simultaneously produce biomass rich in essential fatty acids, Chlorella vulgaris CCAP 211 was continuously grown in a tubular photobioreactor using air alone or air enriched with CO₂ as the sole carbon source. While on one hand, nitrogen‐limited conditions strongly affected biomass growth, conversely, they almost doubled its lipid fraction. Under these conditions using air enriched with 0, 2, 4, 8, and 16% (v/v) CO₂, the maximum biomass concentration was 1.4, 5.8, 6.6, 6.8, and 6.4 g_DB L^?1 on a dry basis, the CO₂ consumption rate 62, 380, 391, 433, and 430 L^?1 day^?1, and the lipid productivity 3.7, 23.7, 24.8, 29.5, and 24.4 mg L^?1 day^?1, respectively. C. vulgaris was able to grow effectively using CO₂‐enriched air, but its chlorophyll a (3.0–3.5 g 100g_DB^?1), chlorophyll b (2.6–3.0 g 100g_DB^?1), and lipid contents (10.7–12.0 g 100g_DB^?1) were not significantly influenced by the presence of CO₂ in the air. Most of the fatty acids in C. vulgaris biomass were of the saturated series, mainly myristic, palmitic, and stearic acids, but a portion of no less than 45% consisted of unsaturated fatty acids, and about 80% of these were high added‐value essential fatty acids belonging to the ω3 and ω6 series. These results highlight that C. vulgaris biomass could be of great importance for human health when used as food additive or for functional food production. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:916–922, 2014