Optimization of carbon and nitrogen sources and growth factors for the production of an aquaculture probiotic (Pseudomonas MCCB 103) using response surface methodology

Aim: To develop a new medium for enhanced production of biomass of an aquaculture probiotic Pseudomonas MCCB 103 and its antagonistic phenazine compound, pyocyanin. Methods and Results: Carbon and nitrogen sources and growth factors, such as amino acids and vitamins, were screened initially in a mineral medium for the biomass and antagonistic compound of Pseudomonas MCCB 103. The selected ingredients were further optimized using a full-factorial central composite design of the response surface methodology. The medium optimized as per the model for biomass contained mannitol (20 g l)1), glycerol (20 g l)1), sodium chloride (5 g l)1), urea (3Æ3 g l)1) and mineral salts solution (20 ml l)1), and the one optimized for the antagonistic compound contained mannitol (2 g l)1), glycerol (20 g l)1), sodium chloride (5Æ1 g l)1), urea (3Æ6 g l)1) and mineral salts solution (20 ml l)1). Subsequently, the model was validated experimentally with a biomass increase by 19% and fivefold increase of the antagonistic compound. Conclusion: Significant increase in the biomass and antagonistic compound production could be obtained in the new media. Significance and Impact of the Study: Media formulation and optimization are the primary steps involved in bioprocess technology, an attempt not made so far in the production of aquaculture probiotics.

Optimization of carbon and nitrogen sources and growth factors for the production of an aquaculture probiotic (Pseudomonas MCCB 103) using response surface methodology

Aim: To develop a new medium for enhanced production of biomass of an aquaculture probiotic Pseudomonas MCCB 103 and its antagonistic phenazine compound, pyocyanin. Methods and Results: Carbon and nitrogen sources and growth factors, such as amino acids and vitamins, were screened initially in a mineral medium for the biomass and antagonistic compound of Pseudomonas MCCB 103. The selected ingredients were further optimized using a full-factorial central composite design of the response surface methodology. The medium optimized as per the model for biomass contained mannitol (20 g l)1), glycerol (20 g l)1), sodium chloride (5 g l)1), urea (3Æ3 g l)1) and mineral salts solution (20 ml l)1), and the one optimized for the antagonistic compound contained mannitol (2 g l)1), glycerol (20 g l)1), sodium chloride (5Æ1 g l)1), urea (3Æ6 g l)1) and mineral salts solution (20 ml l)1). Subsequently, the model was validated experimentally with a biomass increase by 19% and fivefold increase of the antagonistic compound. Conclusion: Significant increase in the biomass and antagonistic compound production could be obtained in the new media. Significance and Impact of the Study: Media formulation and optimization are the primary steps involved in bioprocess technology, an attempt not made so far in the production of aquaculture probiotics

Use of Bioscreen C for growth of Mucor hiemalis in different carbon and nitrogen sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Solubilization of CaHPO4 and AlPO4 by Aspergillus niger in culture media with different carbon and nitrogen sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

d(-)-Lactic Acid Production by Leuconostoc mesenteroides B512 Using Different Carbon and Nitrogen Sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of carbon and nitrogen sources on lipase production by a newly isolated Candida viswanathii strain

Microorganisms can produce lipases with different biochemical characteristics making necessary the screening of new lipase-producing strains for different industrial applications. In this study, 90 microbial strains were screened as potential lipase producers using a sensitive agar plate method with a suitable medium supplemented with Tween 20 and also a liquid culture supplemented with olive oil. The highest cell growth and lipase production for Candida viswanathii were observed in triolein and oleic acid when used as the only pure carbon source. Renewable low-cost triacylglycerols supported the best cell growth, and olive oil was found to be the best inducer for lipase production (19.50 g/L and 58.50 U). The selected conditions for enzyme production were found with yeast extract as nitrogen source and 1.5 % (w/v) olive oil (85.70 U) that resulted in a good cell growth yield (YX/S = 1.234 g/g) and lipase productivity (1.204 U/h) after 72 h of shake-flask cultivation. C. viswanathii lipase presented high hydrolytic activity on esters bonds of triacylglycerols of long-chain, and this strain can be considered an important candidate for future applications in chemical industries. © 2012 Springer-Verlag Berlin Heidelberg and the University of Milan.

Use of Bioscreen C for growth of Mucor hiemalis in different carbon and nitrogen sources

O sistema automatizado Bioscreen C foi utilizado para o crescimento de quatro linhagens de Mucor hiemalis, isoladas do solo da Estação Ecológica de Juréia-Itatins (EEJI), estado de São Paulo, em meios líquidos com uma única fonte de carbono (2%) ou de nitrogênio (1%), pH 5,0, a 25ºC, e agitação de 150rpm por 5 dias. O meio com somente uma única fonte de nitrogênio foi adicionado com 2% de glicose. As leituras de densidade óptica foram realizadas a 540nm, em intervalos de 2h, por cinco dias. Os resultados foram analisados estatisticamente com o Teste de Friedman (alfa = 5%). Os melhores crescimentos foram obtidos com as linhagens M1, M2 e M3, que atingiram o início da fase log em 60 horas de cultivo. As melhores fontes de carbono variaram de acordo com a linhagem estudada, e extrato de levedura provou ser a melhor fonte de nitrogênio para todas as linhagens. Acetato de sódio inibiu o crescimento das quatro linhagens, sendo a M3 a mais afetada. O uso do sistema automatizado foi muito conveniente para as culturas em meio liquido, sendo rápido e automático, constituindo em uma boa técnica para a determinação das condições ambientais ótimas para crescimento de fungos filamentosos.

Stable carbon and nitrogen isotopic compositions of high molecular weight dissolved organic matter from four US estuaries

High molecular weight dissolved organic matter (HMW-DOM) represents an important component of dissolved organic carbon (DOC) in seawater and fresh-waters. In this paper, we report measurements of stable carbon (delta(13)C) isotopic compositions in total lipid, total hydrolyzable amino acid (THAA), total carbohydrate (TCHO) and acid-insoluble "uncharacterized" organic fractions separated from fourteen HMW-DOM samples collected from four U.S. estuaries. In addition, C/N ratio, delta(13) C and stable nitrogen (delta(15)N) isotopic compositions were also measured for the bulk HMW-DOM samples. Our results indicate that TCHO and THAA are the dominant organic compound classes, contributing 33-46% and 13-20% of the organic carbon in HMW-DOM while total lipid accounts for only <2% of the organic carbon in the samples. In all samples. a significant fraction (35-49%) of HMW-DOM was included in the acid-insoluble fraction. Distinct differences in isotopic compositions exist among bulk samples, the compound classes and the acid-insoluble fractions. Values of delta(13)C and delta(15)N measured for bulk HMW-DOM varied from -22.1 to -30.1parts per thousand and 2.8 to 8.9parts per thousand, respectively and varied among the four estuaries studied as well. Among the Compound classes, TCHO was more enriched in C-13 (delta(13)C = -18.5 to -22.8parts per thousand) compared with THAA (delta(13)C = -20.0 to -29.6parts per thousand) and total lipid (delta(13)C = -25.7 to -30.7parts per thousand). The acid-insoluble organic fractions, in general, had depleted C-13 values (delta(13)C = -23.0 to -34.4parts per thousand). Our results indicate that the observed differences in both delta(13)C and delta(15)N were mainly due to the differences in sources of organic matter and nitrogen inputs to these estuaries in addition to the microbial processes responsible for isotopic fractionation among the compound classes. Both terrestrial sources and local sewage inputs contribute significantly to the HMW-DOM pool in the estuaries studied and thus had a strong influence on its isotopic signatures. Copyright (C) 2004 Elsevier Ltd.

Analysis of diets of upland buzzards using stable carbon and nitrogen isotopes

We measured the stable carbon and nitrogen isotope ratios for muscles of the upland buzzards (Buteo hemilasius) and their potential food sources, plateau pikas (Ochotona curzoniae), Qinghai voles (Lasiopodomys fuscus), plateau zokors (Myospalax fontanierii), and several passerine bird species at the alpine meadow in Maduo county, Guoluo prefecture of Qinghai province, People's Republic of China, to provide diet information of upland buzzards, highlighting different diet composition of upland buzzards exposed to different locations. The results demonstrated that stable carbon isotope ratios of upland buzzards, passerine birds, plateau pikas, plateau zokors, and Qinghai voles were -24.42 +/- 0.25parts per thousand, -22.89 +/- 1.48parts per thousand, -25.30 +/- 1.47parts per thousand, -25.78 +/- 0.22parts per thousand, and -25.41 +/- 0.01parts per thousand, respectively, and stable nitrogen isotope ratios were 7.89 +/- 0.38parts per thousand, 8.37 +/- 2.05parts per thousand, 5.83 +/- 1.10parts per thousand, 5.23 +/- 0.34parts per thousand, and 8.86 +/- 0.06parts per thousand, respectively. Fractionation of stable carbon and nitrogen isotope ratios between upland buzzards and their food were 1.03parts per thousand and 2.11parts per thousand, respectively. Based on mass balance principle of stable isotopes and the Euclidean distance mixing model, upland buzzards depended mainly on plateau pikas as food (74.56%). Plateau zokors, Qinghai voles, and passerine birds only contributed a small proportion (25.44%) to diets of upland buzzards. The results were closely accordant with analyses of stomach contents and food pellets, which firmly supported the feasibility of using stable carbon and nitrogen isotope ratios to investigate diet information of upland buzzards. Another study based on stable carbon isotopes showed that upland buzzards living in the Haibei prefecture (another prefecture located in the southeast Qinghai province) mainly preyed on passerine birds (64.96% or more) as food supply. We were alarmed by the preliminary results that widespread poisoning activities of small mammals could reshape the food composition of upland buzzards, influencing the stability and sustainability of the alpine meadow. Bio-control on rodent pests should be carried out rather than the chemical measures.

Organo-mineral complexation alters carbon and nitrogen cycling in stream microbial assemblages

Inland waters are of global biogeochemical importance receiving carbon inputs of ~ 4.8 Pg C y^-1. Of this 12 % is buried, 18 % transported to the oceans, and 70 % supports aquatic secondary production. However, the mechanisms that determine the fate of organic matter (OM) in these systems are poorly defined. One important aspect is the formation of organo-mineral complexes in aquatic systems and their potential as a route for OM transport and burial vs. their use potential as organic carbon (C) and nitrogen (N) sources. Organo-mineral particles form by sorption of dissolved OM to freshly eroded mineral surfaces and may contribute to ecosystem-scale particulate OM fluxes. We tested the availability of mineral-sorbed OM as a C & N source for streamwater microbial assemblages and streambed biofilms. Organo-mineral particles were constructed in vitro by sorption of ¹³C:¹⁵N-labelled amino acids to hydrated kaolin particles, and microbial degradation of these particles compared with equivalent doses of ¹³C:¹⁵N-labelled free amino acids. Experiments were conducted in 120 ml mesocosms over 7 days using biofilms and streamwater sampled from the Oberer Seebach stream (Austria), tracing assimilation and mineralization of ¹³C and ¹⁵N labels from mineral-sorbed and dissolved amino acids.Here we present data on the effects of organo-mineral sorption upon amino acid mineralization and its C:N stoichiometry. Organo-mineral sorption had a significant effect upon microbial activity, restricting C and N mineralization by both the biofilm and streamwater treatments. Distinct differences in community response were observed, with both dissolved and mineral-stabilized amino acids playing an enhanced role in the metabolism of the streamwater microbial community. Mineral-sorption of amino acids differentially affected C & N mineralization and reduced the C:N ratio of the dissolved amino acid pool. The present study demonstrates that organo-mineral complexes restrict microbial degradation of OM and may, consequently, alter the carbon and nitrogen cycling dynamics within aquatic ecosystems.

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Carbon and nitrogen stock and fluxes in coastal Atlantic Forest of southeast Brazil: potential impacts of climate change on biogeochemical functioning

The Atlantic Forest is one of the most important biomes of Brazil. Originally covering approximately 1.5 million of km(2), today this area has been reduced to 12% of its original size. Climate changes may alter the structure and the functioning of this tropical forest. Here we explore how increases in temperature and changes in precipitation distribution could affect dynamics of carbon and nitrogen in coastal Atlantic Forest of the southeast region of Brazil The main conclusion of this article is that the coastal Atlantic Forest has high stocks of carbon and nitrogen above ground, and especially, below ground. An increase in temperature may transform these forests from important carbon sinks to carbon sources by increasing loss of carbon and nitrogen to the atmosphere. However, this conclusion should be viewed with caution because it is based on limited information. Therefore, more studies are urgently needed to enable us to make more accurate predictions.

A trophic study of the sympatric Amazonian freshwater turtles Podocnemis unifilis and Podocnemis expansa (Testudines, Podocnemidae) using carbon and nitrogen stable isotope analyses

The Yellow-spotted River Turtle (Podocnemis unifilis Troschel, 1848) and the South American River Turtle (Podocnemis expansa (Schweigger, 1812)) are two turtles species that are widely distributed and have ecological, economic, and cultural importance in the Amazon basin. Although sympatric regarding most of their distribution, few studies have addressed the coexistence of these two species. To examine this, we analyzed the trophic level and the primary carbon source from the diets of both species in Baixo Araguaia, Tocantins, Brazil, using stable isotope analyses of carbon (delta C-13) and nitrogen (delta N-15). We also verified possible intraspecific variations (related to sex and body mass) in the trophic levels and primary carbon sources of their diets. Podocnemis unifilis had higher values of delta N-15 than P. expansa, averaging 7.59 parts per thousand and 5.06 parts per thousand, respectively, a difference which may indicate a possible trophic change owing to exploiting different food resources. No differences were found between the two species in relation to delta C-13 (mean values of -26.2 parts per thousand and -26.1 parts per thousand, respectively). The similarity between delta C-13 values suggests that the sources of their basal feeding are the same, consisting mainly of C-3 plants. There was no intraspecific variation in the values of delta C-13 and delta N-15.

Interaction of sulfate assimilation with carbon and nitrogen metabolism in Lemna minor

Cysteine synthesis from sulfide andO-acetyl-L-serine (OAS) is a reaction interconnecting sulfate, nitrogen, and carbon assimilation. UsingLemna minor, we analyzed the effects of omission of CO2 from the atmosphere and simultaneous application of alternative carbon sources on adenosine 5′-phosphosulfate reductase (APR) and nitrate reductase (NR), the key enzymes of sulfate and nitrate assimilation, respectively. Incubation in air without CO2 led to severe decrease in APR and NR activities and mRNA levels, but ribulose-1,5-bisphosphate carboxylase/oxygenase was not considerably affected. Simultaneous addition of sucrose (Suc) prevented the reduction in enzyme activities, but not in mRNA levels. OAS, a known regulator of sulfate assimilation, could also attenuate the effect of missing CO2 on APR, but did not affect NR. When the plants were subjected to normal air after a 24-h pretreatment in air without CO2, APR and NR activities and mRNA levels recovered within the next 24 h. The addition of Suc and glucose in air without CO2 also recovered both enzyme activities, with OAS again influenced only APR.35SO4 2− feeding showed that treatment in air without CO2 severely inhibited sulfate uptake and the flux through sulfate assimilation. After a resupply of normal air or the addition of Suc, incorporation of 35S into proteins and glutathione greatly increased. OAS treatment resulted in high labeling of cysteine; the incorporation of 35S in proteins and glutathione was much less increased compared with treatment with normal air or Suc. These results corroborate the tight interconnection of sulfate, nitrate, and carbon assimilation.

Carbon and Nitrogen concentrations and isotopic composition of ODP Site 170-1040 and Leg 205 sites

We determined the C and N concentrations and isotopic compositions of sediments in the prism sampled during Ocean Drilling Program Legs 170 and 205 offshore Costa Rica, with the goals of evaluating sediment sources and extents of diagenesis and identifying any effects of infiltrating fluids on the sedimentary C and N. The sediments from Leg 170 Site 1040 contain 0.85-1.96 wt% total organic carbon (TOC) with Vienna Peedee belemnite (VPDB) d13CVPDB from -26.3 per mil to -22.5 per mil, and 832-2221 ppm total nitrogen (TN) with d15Nair from +3.5 per mil to +6.6 per mil. Sediment TN concentrations and d15N values show dramatic downhole increases within the uppermost 130 m of the section and more gradual downhole decreases from 130 meters below seafloor (mbsf) to the base of the décollement at ~370 mbsf. Concentrations and isotopic compositions of TOC are relatively uniform within the entire section, showing some minor perturbation within the décollement zone. In the uppermost 100 m, upsection increases in TN concentrations at constant TOC concentrations produce significant increases in atomic TOC/TN ratios from ~8 to ~18. Carbonate (calcite) contents in the wedge sediments are generally low (<4 wt%), but the d13C and Vienna standard mean ocean water (VSMOW) d18OVSMOW values vary significantly from -26.1 per mil to +4.1 per mil and from +30.0 per mil to +35.3 per mil, respectively. Concentrations and isotopic compositions of TOC and TN for sediments from Leg 205 Sites 1254 and 1255 overlap well with C-N data for sediments from the same depth intervals obtained during Leg 170 at Site 1040.