Structure determination of partially deuterated carotenoids from intrinsically labeled vegetables by HPLC-MS and 1H NMR

The structures of biosynthetic deuterated carotenoids in labeled vegetables were investigated: (all-E)-lutein and (all-E)-beta-carotene from spinach, and (all-E)-beta-carotene and (all-E)-alpha-carotene from carrots. The vegetables were grown hydroponically using a nutrient solution enriched with deuterium oxide (D(2)O) and were extracted using matrix solid-phase dispersion (MSPD). Deuterium enrichment in the carotenoid molecules was determined by liquid chromatography-mass spectrometry (LC-MS). (all-E)-Lutein and (all-E)-beta-carotene in spinach showed partial deuteration from (2)H(1) to (2)H(12), with the abundance maximum at (2)H(5). (all-E)-beta-Carotene and (all-E)-alpha-carotene from carrots showed partial deuteration from (2)H(1) to (2)H(17), with the abundance maximum at (2)H(11). The (1)H NMR spectra of the four deuterated carotenoids showed additional signals for all methyl groups and decreased signal intensity for the olefinic protons and the methylene protons in the ring. These differences are due to isotopic effects and are based on the substitution of protons by deuterium atoms. The deuteration was distributed randomly throughout the carotenoid molecules.     

Capillary electrophoresis and high-performance liquid chromatography determination of polyglutamyl 5-methyltetrahydrofolate forms in citrus products

The 5-methyltetrahydrofolate (5mTHF) polyglutamates in citrus products were analyzed by capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). Folate species were purified from citrus products and concentrated from 2- to 100-fold using combined folate-affinity chromatography and C18 extraction. Seven polyglutamyl 5mTHFs were found in most not-from-concentrate (NFC) orange juices (OJ) in total amounts of approximately 1 nmol/mL, with varying distributions of individual polyglutamates. Folate amounts and distributions were also measured in orange fractions, single-strength OJ from concentrate, NFC grapefruit juice, and citrus peel molasses. Models containing ascorbic acid had folate thermal degradation rates one-seventh that of models without ascorbic acid. Pasteurization studies demonstrated that folate loss was <2% for commercial OJ pasteurization conditions (i.e., 93 degrees C for 5 s, 88 degrees C for 15 s, and 82 degrees C for 30 s). Both methods were precise, reproducible, and potentially faster than traditional analytical procedures requiring enzymatic deconjugation and microbial assays.     

Control of nitrogen export from watersheds by headwater streams

A comparative (15)N-tracer study of nitrogen dynamics in headwater streams from biomes throughout North America demonstrates that streams exert control over nutrient exports to rivers, lakes, and estuaries. The most rapid uptake and transformation of inorganic nitrogen occurred in the smallest streams. Ammonium entering these streams was removed from the water within a few tens to hundreds of meters. Nitrate was also removed from stream water but traveled a distance 5 to 10 times as long, on average, as ammonium. Despite low ammonium concentration in stream water, nitrification rates were high, indicating that small streams are potentially important sources of atmospheric nitrous oxide. During seasons of high biological activity, the reaches of headwater streams typically export downstream less than half of the input of dissolved inorganic nitrogen from their watersheds.     

Short-term effects of aglime on inorganic- and organic-derived CO2 emissions from two acid soils amended with an ammonium-based fertiliser

Purpose

Aglime application can promote carbon dioxide (CO₂) emissions from acid soils. However, the controlling mechanisms are still poorly understood, particularly the role of fertiliser-ammonium oxidation. This study therefore assessed the effects of aglime on soil inorganic C (SIC)– and soil organic C (SOC)–derived CO₂ emissions from acid soils amended with ammonium.

Materials and methods

Ammonium at three N rates [0% (A0), 0.005% (A1), and 0.2% (A2) w/w] and labelled aglime (Ca¹³CO_3,¹³C 5.94% aa) at three rates [0% (L1), 0.067% (L1), and 0.392% (L2) w/w] were applied to two contrasting acid soils (Nariva series, Mollic Fluvaquents; and Piarco series, Typic Kanhaplaquults) and incubated in 1-l media bottles for 23 days. A calcareous soil (Princes Town series, Aquentic Eutrudepts, carbonate δ¹³C of ??4.79‰) was included as a control that only received ammonium at the three rates.

Results and discussion

The application of ammonium at the A2 rate significantly (p?<?0.05) increased cumulative SIC-CO₂ emissions by 15.8 and 27.1% in comparison to the A0 rate for the Nariva and Piarco soils, respectively, when they were limed at the L2 rate. The lower rate of ammonium (A1), however, had no effect on these emissions, which suggests that enough acidity may not have been generated at this rate to significantly enhance the release of SIC-CO₂. Furthermore, no effect of ammonium rates was observed on SIC-CO₂ emissions from the calcareous soil, which refutes the hypothesis that this amendment plays a greater role in regulating these emissions from calcareous soils compared with acid soils. Also, in contradiction to another hypothesis, the aglime-induced priming effect on SOC decomposition was more apparent in the low-C Piarco soil. This effect was also significantly (p?<?0.05) greater at the L2 rate (above the lime requirement for Piarco), which demonstrates the negative impact that over-liming could have on the sequestration of C in this soil. Our results also showed that ammonium addition may also help to reduce the magnitude of the aglime-induced priming effect in the Piarco soil when it is not over-limed.

Conclusions

Overall, the findings of this study suggest that ammonium fertiliser broadcast at conventional rates may not serve as a significant regulator of SIC-CO₂ emissions from highly to moderately acidic soils amended with aglime. Our findings also indicate a need to consider nitrogen management as an important factor regulating the effects of aglime on SOC-CO₂ emissions.

     

The physical state and plasma biochemical profile of young calves on arrival at a slaughter plant

AIM: To determine the welfare of calves arriving at a slaughter plant by assessing their physical state and plasma biochemical profile. METHODS: Calves (n=7,169) aged 4-10 days, were examined immediately after they were unloaded from trucks (n=20) and categorised on physical grounds as being in an 'unacceptable', 'marginal' or 'acceptable' state. 'Unacceptable' calves were lying down, unable to walk, extremely weak or seriously injured. 'Marginal' calves had a wet umbilicus, were hollow sided, apparently immature, or weak and slow and unsteady on their feet. 'Acceptable' calves were strong, walked freely, round-sided, bright and alert and had a dry umbilicus. After unloading, 27 'unacceptable', 306 'marginal' and 289 'acceptable' calves were weighed, examined clinically and a blood sample was taken from each. A second blood sample was taken from 118 of these calves, 12-15 hours later. The packed cell volume (PCV) and plasma concentrations of glucose, beta-hydroxybutyrate (BHB), total plasma proteins (TPP), gamma-glutamyl transferase (GGT), urea, creatine kinase (CK) and lactate were measured. RESULTS: Only 27 of the 7,169 calves examined (0.4%) were categorised as being in an 'unacceptable' state on arrival and these were euthanased shortly after unloading. Generally, the plasma biochemical profiles of the 'acceptable' calves were similar to those of 'marginal' animals. However the glucose concentration of 'marginal' calves was lower than that of 'acceptable' calves, suggesting that they had lower energy reserves. After 12-15 h in lairage, plasma glucose concentration decreased and the BHB concentration increased in both 'acceptable' and 'marginal' calves compared with concentrations measured on arrival. CONCLUSIONS: The percentage of calves presented for slaughter in an 'unacceptable' state was small (0.4%). However, 4% of calves were classified as 'marginal' and failed to meet all the requirements of the code of recommendations and minimum standards governing the welfare of young calves in New Zealand. The categorisation of calves by a physical assessment was supported by results of plasma biochemistry. As a result, plasma biochemical profiles are considered unnecessary for making decisions regarding the welfare status of calves arriving at a slaughter plant; an assessment of their physical state is sufficient.     

Evaluation of a portocaval venograft and ameroid ring for the occlusion of intrahepatic portocaval shunts in dogs

OBJECTIVE:To evaluate the use of a portocaval venograft and ameroid constrictor in the surgical management of intrahepatic portosystemic shunts (PSS). STUDY DESIGN: Prospective, clinical study. Animal Population: Ten client-owned dogs with intrahepatic PSS. METHODS: Portal pressure was measured after temporary suture occlusion of the intrahepatic PSS. In dogs with an increase in portal pressure greater than 8 mm Hg, a single extrahepatic portocaval shunt was created using a jugular vein. An ameroid ring was placed around the venograft and the intrahepatic PSS was attenuated. Transcolonic pertechnetate scintigraphy was performed before surgery, 5 days after surgery, and 8 to 10 weeks after surgery. Dogs with continued portosystemic shunting were evaluated further by laparotomy or portography. Clinical outcome and complications were recorded. RESULTS: Mean (+/- SD) portal pressure increased from 6 +/- 3 to 19 +/- 6 mm Hg with PSS occlusion; in all 10 dogs, the increase in portal pressure was greater than 8 mm Hg. There were no intraoperative complications, and, after creation of the portocaval shunt, the intrahepatic PSS could be completely ligated in 8 of 10 dogs. The final portal pressure was 9 +/- 4 mm Hg. Postoperative complications included coagulopathy and death (1 dog), ascites (3 dogs), and incisional discharge (3 dogs). Five of 8 dogs had continued portosystemic shunting at 8 to 10 weeks after surgery. Multiple extrahepatic PSS were demonstrated in 4 of these dogs. Clinical outcome was excellent in all 9 surviving dogs. CONCLUSIONS AND CLINICAL SIGNIFICANCE: The surgical technique resulted in a high incidence of multiple extrahepatic PSS. Short-term clinical results were promising, but long-term outcome must be evaluated further.