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

Carbon and nitrogen emissions from stored manure and cropped fields in irrigated mountain oases of Oman

Little is known about gaseous carbon (C) and nitrogen (N) emissions from traditional terrace agriculture in irrigated high mountain agroecosystems of the subtropics. In an effort towards filling this knowledge gap measurements of carbon dioxide (CO_2), methane (CH_4), ammonia (NH_3) and dinitrous oxide (N_2O) were taken with a mobile photoacoustic infrared multi-gas monitor on manure-filled PE-fibre storage bags and on flood-irrigated untilled and tilled fields in three mountain oases of the northen Omani Al Jabal al Akhdar mountains. During typical 9-11 day irrigation cycles of March, August and September 2006 soil volumetric moisture contents of fields dominated by fodder wheat, barley, oats and pomegranate ranged from 46-23%. While manure incorporation after application effectively reduced gaseous N losses, prolonged storage of manure in heaps or in PE-fibre bags caused large losses of C and N. Given the large irrigation-related turnover of organic C, sustainable agricultural productivity of oasis agriculture in Oman seems to require the integration of livestock which allows for several applications of manure per year at individual rates of 20 t dry matter ha^−1.

Gaseous and leaching losses of carbon and nitrogen in irrigated organic farming of a coastal oasis in Oman

In der Dissertation wurden die Effekte verschiedener C/N-Verhältnisse und verschiedener Verhältnisse von strukturellen zu löslichen Kohlenhydraten (NDF/SC) von Dung, der in bewässerten Gemüsekulturen im Norden Omans appliziert wurde, untersucht. Im auf sandigen Böden durchgeführten Experiment wurden zwei Büffeldungvarianten zum einen mit einem C/N-Verhältnis von 19 und einem NDF/SC-Verhältnis von 17 (ORG1) und zum anderen mit einem C/N-Verhältnis von 25 und einem NDF/SC-Verhältnis von 108 (ORG2) verwendet. Das relevante faktorielle Anbausystem war eine zweijährige Rotation, bestehend aus Rettich gefolgt von Blumenkohl und Karotte. Eine signifikante Zunahme der Erträge, des Sproßdurchmessers und der Pflanzenhöhe von Blumenkohl (P<0,001) sowie der Konzentration von Askorbinsäure in den Wurzeln von Rettich (P<0,01) mit erhöhter Verfügbarkeit von N, P und K von ORG2 über ORG1 bis hin zur Mineraldünger-Kontrollbehandlung (MIN) konnte festgestellt werden. Innerhalb von 260 Tagen wurden für die gesamte Anbauperiode mit einem photoakustischen Infrarot-Multigasmonitor und einer damit verbundenen Haube bodenbürtige Gasemissionen gemessen. Die errechneten Nettobilanzen zeigten Überschüsse von N und P, welche von Defiziten für K begleitet waren. Die Kohlenstoff Nettobilanzen waren während des Untersuchungszeitraums negativ oder nicht konsistent. Die Ergebnisse zeigen, dass unter extremen klimatischen Bedingungen bewässerter sandiger Böden organische Kultivierung zuerst durch den Kohlenstoffgehalt von Dung und Boden und erst dann durch die applizierten Mengen an N, P und K limitiert wird. Es konnte festgestellt werden, dass Gasemissionen den größten Teil der N und C Verluste von bewässerten sandigen Böden im Norden Omans darstellen. Die Reduzierung von Treibhausgasen und Sickerverlusten sollte weiterhin im Fokus zukünftiger Untersuchungen stehen, um zur Entwicklung von nachhaltigen organischen Anbausystemen im Oman und anderen ariden tropischen Ländern beizutragen.

Carbon and nitrogen isotopic ratios of urine and faeces as novel nutritional biomarkers of meat and fish intake

Purpose Meat and fish consumption are associated with changes in the risk of chronic diseases. Intake is mainly assessed using self-reporting, as no true quantitative nutritional biomarker is available. The measurement of plasma fatty acids, often used as an alternative, is expensive and time-consuming. As meat and fish differ in their stable isotope ratios, δ13C and δ15N have been proposed as biomarkers. However, they have never been investigated in controlled human dietary intervention studies. Objective In a short-term feeding study, we investigated the suitability of δ13C and δ15N in blood, urine and faeces as biomarkers of meat and fish intake. Methods The dietary intervention study (n = 14) followed a randomised cross-over design with three eight-day dietary periods (meat, fish and half-meat–half-fish). In addition, 4 participants completed a vegetarian control period. At the end of each period, 24-h urine, fasting venous blood and faeces were collected and their δ13C and δ15N analysed. Results There was a significant difference between diets in isotope ratios in faeces and urine samples, but not in blood samples (Kruskal–Wallis test, p < 0.0001). In pairwise comparisons, δ13C and δ15N were significantly higher in urine and faecal samples following a fish diet when compared with all other diets, and significantly lower following a vegetarian diet. There was no significant difference in isotope ratio between meat and half-meat–half-fish diets for blood, urine or faecal samples. Conclusions The results of this study show that urinary and faecal δ13C and δ15N are suitable candidate biomarkers for short-term meat and fish intake.

Carbon and nitrogen stable isotope ratio analysis of freshwater, brackish and marine fish from Belgian archaeological sites (1st and 2nd millennium AD)

Carbon and nitrogen stable isotope ratios were measured in 157 fish bone collagen samples from 15 different archaeological sites in Belgium which ranged in ages from the 3rd to the 18th c. AD. Due to diagenetic contamination of the burial environment, only 63 specimens produced results with suitable C:N ratios (2.9–3.6). The selected bones encompass a wide spectrum of freshwater, brackish, and marine taxa (N = 18), and this is reflected in the δ13C results (−28.2‰ to −12.9%). The freshwater fish have δ13C values that range from −28.2‰ to −20.2‰, while the marine fish cluster between −15.4‰ and −13.0‰. Eel, a catadromous species (mostly living in freshwater but migrating into the sea to spawn), plots between −24.1‰ and −17.7‰, and the anadromous fish (living in marine environments but migrating into freshwater to spawn) show a mix of freshwater and marine isotopic signatures. The δ15N results also have a large range (7.2‰ to 16.7‰) indicating that these fish were feeding at many different trophic levels in these diverse aquatic environments. The aim of this research is the isotopic characterization of archaeological fish species (ecology, trophic level, migration patterns) and to determine intra-species variation within and between fish populations differing in time and location. Due to the previous lack of archaeological fish isotope data from Northern Europe and Belgium in particular, these results serve as an important ecological backdrop for the future isotopic reconstruction of the diet of human populations dating from the historical period (1st and 2nd millennium AD), where there is zooarchaeological and historical evidence for an increased consumption of marine fish.

Serum carbon and nitrogen stable isotopes as potential biomarkers of dietary intake and their relation with incident type 2 diabetes: the EPIC-Norfolk study

Background: Stable-isotope ratios of carbon (13C/12C, expressed as δ13C) and nitrogen (15N/14N, or δ15N) have been proposed as potential nutritional biomarkers to distinguish between meat, fish, and plant-based foods. Objective: The objective was to investigate dietary correlates of δ13C and δ15N and examine the association of these biomarkers with incident type 2 diabetes in a prospective study. Design: Serum δ13C and δ15N (‰) were measured by using isotope ratio mass spectrometry in a case-cohort study (n = 476 diabetes cases; n = 718 subcohort) nested within the European Prospective Investigation into Cancer and Nutrition (EPIC)–Norfolk population-based cohort. We examined dietary (food-frequency questionnaire) correlates of δ13C and δ15N in the subcohort. HRs and 95% CIs were estimated by using Prentice-weighted Cox regression. Results: Mean (±SD) δ13C and δ15N were −22.8 ± 0.4‰ and 10.2 ± 0.4‰, respectively, and δ13C (r = 0.22) and δ15N (r = 0.20) were positively correlated (P < 0.001) with fish protein intake. Animal protein was not correlated with δ13C but was significantly correlated with δ15N (dairy protein: r = 0.11; meat protein: r = 0.09; terrestrial animal protein: r = 0.12, P ≤ 0.013). δ13C was inversely associated with diabetes in adjusted analyses (HR per tertile: 0.74; 95% CI: 0.65, 0.83; P-trend < 0.001], whereas δ15N was positively associated (HR: 1.23; 95% CI: 1.09, 1.38; P-trend = 0.001). Conclusions: The isotope ratios δ13C and δ15N may both serve as potential biomarkers of fish protein intake, whereas only δ15N may reflect broader animal-source protein intake in a European population. The inverse association of δ13C but a positive association of δ15N with incident diabetes should be interpreted in the light of knowledge of dietary intake and may assist in identifying dietary components that are associated with health risks and benefits.

Effects of diet change on carbon and nitrogen stable-isotope ratios in blood cells and plasma of the long-nosed bandicoot (Perameles nasuta)

Carbon (C) and nitrogen (N) stable isotopes offer a powerful tool for assessing the extent of tissue assimilation of dietary components. However, the method relies on knowledge of diet-tissue isotopic discrimination and how quickly diet shifts become apparent in various tissues. In the present study, blood plasma and blood cells, tissues that are easily obtained under field conditions, were used to validate the stable isotope method over a period of 4-5 weeks using captive long-nosed bandicoots (Perameles nasuta). Diet-tissue discrimination effects appeared to be small. For C, derived diet-tissue isotopic discriminations were 1.4‰ for blood plasma and -0.2‰ for blood cells. For N the values were 2.8‰ and 2.1‰, respectively, and were independent of the nitrogen content of the food. C and N turnover measurements in the blood plasma and cells of the bandicoots indicated that blood plasma provides dietary information integrated over a period of ∼3 weeks, whereas blood cells give an impression of the assimilated diet over a period of as much as half a year. These turnover rates were low compared with the little information available for birds and eutherian mammals, and probably relate to the typically low metabolic rate of marsupials.

Metabolite profiling reveals distinct changes in carbon and nitrogen metabolism in phosphate-deficient barley plants (Hordeum vulgare L.)

Plants modify metabolic processes for adaptation to low phosphate (P) conditions. Whilst transcriptomic analyses show that P deficiency changes hundreds of genes related to various metabolic processes, there is limited information available for global metabolite changes of P-deficient plants, especially for cereals. As changes in metabolites are the ultimate ‘readout’ of changes in gene expression, we profiled polar metabolites from both shoots and roots of P-deficient barley (Hordeum vulgare) using gas chromatography–mass spectrometry (GC-MS). The results showed that mildly P-deficient plants accumulated di- and trisaccharides (sucrose, maltose, raffinose and 6-kestose), especially in shoots. Severe P deficiency increased the levels of metabolites related to ammonium metabolism in addition to di- and trisaccharides, but reduced the levels of phosphorylated intermediates (glucose-6-P, fructose-6-P, inositol-1-P and glycerol-3-P) and organic acids (α-ketoglutarate, succinate, fumarate and malate). The results revealed that P-deficient plants modify carbohydrate metabolism initially to reduce P consumption, and salvage P from small P-containing metabolites when P deficiency is severe, which consequently reduced levels of organic acids in the tricarboxylic acid (TCA) cycle. The extent of the effect of severe P deficiency on ammonium metabolism was also revealed by liquid chromatography–mass spectrometry (LC-MS) quantitative analysis of free amino acids. A sharp increase in the concentrations of glutamine and asparagine was observed in both shoots and roots of severely P-deficient plants. Based on these data, a strategy for improving the ability of cereals to adapt to low P environments is proposed that involves alteration in partitioning of carbohydrates into organic acids and amino acids to enable more efficient utilization of carbon in P-deficient plants.

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)

The traceability of animal meals in layer diets as detected by stable carbon and nitrogen isotope analyses of eggs

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

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.

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)