Development of an indirect competitive ELISA for flumequine residues in raw milk using chicken egg yolk antibodies

To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA-flumequine) and to cationized ovalbumin (cOVA-flumequine). For the immunization of chickens, cBSA-flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA-flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 microg/kg). No cross-reactivity was noticed with various related quinolones.     

The silica balance in the world ocean: a reestimate

The net inputs of silicic acid (dissolved silica) to the world ocean have been revised to 6.1 +/- 2.0 teramoles of silicon per year (1 teramole = 10(12) moles). The major contribution (about 80 percent) comes from rivers, whose world average silicic acid concentration is 150 micromolar. These inputs are reasonably balanced by the net ouputs of biogenic silica of 7.1 +/- 1.8 teramoles of silicon per year in modern marine sediments. The gross production of biogenic silica (the transformation of dissolved silicate to particulate skeletal material) in surface waters was estimated to be 240 +/- 40 teramoles of silicon per year, and the preservation ratio (opal accumulation in sediment/gross production in surface waters) averages 3 percent. In the world ocean the residence time of silicon, relative to total biological uptake in surface waters, is about 400 years.     

Positive nitrogen balance of <Emphasis Type="Italic">Acacia mangium</Emphasis> woodlots as fallows in the Philippines based on <Superscript>15</Superscript>N natural abundance data of N<Subscript>2</Subscript> fixation

Nitrogen inputs from biological nitrogen fixation contribute to productivity and sustainability of agroforestry systems but they need to be able to offset export of N when trees are harvested. This study assessed magnitudes of biological nitrogen fixation (natural ¹⁵N abundance) and N balance of Acacia mangium woodlots grown in farmer’s fields, and determined if N₂ fixation capacity was affected by tree age. Tree biomass, standing litter, understory vegetation and soil samplings were conducted in 15 farmer’s fields growing A. mangium as a form of sequential agroforestry in Claveria, Misamis Oriental, Philippines. The trees corresponded to ages of 4, 6, 8, 10 and 12 years, and were replicated three times. Samples from different plant parts and soils (0–100 cm) were collected and analyzed for δ¹⁵N and nutrients. The B-value, needed as a reference of isotopic discrimination when fully reliant on atmospheric N, was generated by growing A. mangium in an N₂-free sand culture in the glasshouse. Isotopic discrimination occurring during N₂ fixation and metabolic processes indicated variation of δ¹⁵N values in the order of nodules > old leaves > young leaves > stems > litterfall and roots of the trees grown in the field, with values ranging from −0.8 to 3.5‰ except nodules which were enriched and significantly different from other plant parts (P < 0.0001). Isotopic discrimination was not affected by tree age (P > 0.05). Plants grown in N free sand culture exhibited the same pattern of isotopic discrimination as plants grown in the field. The estimated B-value for the whole plant of A. mangium was −0.86‰. Mature tree stands of 12 years accumulated up to 1994 kg N ha⁻¹ in aboveground biomass. Average proportion of N derived from N₂ fixation of A. mangium was 54% (±22) and was not affected by age (P > 0.05). Average yearly quantities of N₂ fixed were 128 kg N ha⁻¹ in above-ground biomass amounting to 1208 kg N fixed ha⁻¹ over 12 years. Harvest of 12-year old trees removed approximately 91% of standing aboveground biomass from the site as timber and fuel wood. The resulting net N balance was +151 kg N ha⁻¹ derived from remaining leaves, twigs, standing litter, and +562 kg N ha⁻¹ when tree roots were included in the calculation. The fast growing A. mangium appears to be a viable fallow option for managing N in these systems. However, other nutrients have to be replaced by using part of the timber and fuel wood sales to compensate for large amounts of nutrient removed in order for the system to be sustainable.     

Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Comparative energy and economic analyses of conventional and System of Rice Intensification (SRI) methods of rice production in Thai Nguyen Province,Vietnam

The consumption of energy inputs in agricultural production has been increasing rapidly during the past decades. However, given the limitations and costs of non-renewable energy, increasing production while using the least energy possible has become a major concern of most nations. Prompted by this concern, we conducted a face-to-face survey of 90 farming households in Thai Nguyen Province, Vietnam, to find out how energy is being used in agriculture and, specifically, in their rice production. Through analysis of energy input–output balances, combined with economic efficiency analysis, a comparison was made of conventional and SRI methods of rice production. The study found that applying the SRI method can save around 23% of energy inputs, while increasing energy outputs by 11%. Economic benefits per hectare also rise by more than 8 million dong (USD 364) compared to those under the conventional cultivation system. The study also showed conflicts between the energy and economic balances for manual compared with machine ploughing operations. This study contributes to providing an overview of energy consumption in rice cultivation at the household level. Its findings can help stakeholders to assess current policies and make better decisions on the uses of energy in agricultural production. In addition, the comprehensive approach taken here to analysing energy use and efficiency could expand the analysis and comparison of energy uses at sectoral or activity level—still a new field in Vietnam and many other countries.     

Induced expression of selected plant defence related genes in pot azalea, <Emphasis Type="Italic">Rhododendron simsii</Emphasis> hybrid

A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and RsJMT) and salicylic acid (SA) (RsPAL and RsICS). Furthermore, RsPPO, a putative marker gene for oxidative stress response was successfully cloned from R. simsii. A CTAB-based extraction protocol was optimized to assure excellent RNA quality for subsequent RT-qPCR analysis. The RT-qPCR protocol was extensively tested and RsRG7 and RsRG14 were selected as reference genes from a geNorm pilot study. Validation of the marker genes was done after application with elicitors [methyl jasmonate (MeJA), coronatine, β-aminobutyric acid and acibenzolar-Smethyl] or wounding. Both 100 μM MeJA and 0.1 μM coronatine had a significant effect on the expression of all marker genes. Foliar application of MeJA on the shoots resulted in a significantly earlier response when compared to root application and subsequent sampling of the shoots. Expression patterns after MeJA treatment were generally the same in six R. simsii genotypes: ‘Nordlicht’, ‘Elien’, ‘Aiko Pink’, ‘Michelle Marie’, ‘Mevrouw Gerard Kint’ and ‘Sachsenstern’. Wounding resulted in the same expression patterns as MeJA treatment except for RsJMT. None of the genotypes showed a significant induction of the latter gene 6 h upon wounding. Findings of these experiments indicated that the tolerant genotype ‘Elien’ has low basal expression levels of RsPPO. This might be the first step towards the breeding of mite-tolerant genotypes.     

A review of carbon isotopes of phytoliths: implications for phytolith-occluded carbon sources

Journal of Soils and Sediments - Phytolith-occluded carbon (PhytOC) is mainly derived from the products of photosynthesis, which can be preserved in soils and sediments for hundreds-to-thousands of...     

Effects of porcine relaxin on induced parturition in beef heifers

Two-year-old crossbred beef heifers were used to test the effects of porcine relaxin (pRelaxin) alone, or in combination with dexamethasone, on the induction of parturition, the incidence of dystocia, and retained placentas. Effects of treatment on pelvic area, postpartum interval, milk production, colostrum quality, calf birth weight, calf vigor, and calf performance were also evaluated. On Day 275 of gestation, heifers from two fetal-sire groups were randomly assigned to one of four groups in a 2 × 2 factorial design and received: no treatment (controls, N = 19), 20 mg of dexamethasone intramuscularly (im) (n = 22), 5 mg of pRelaxin (3,000 U/mg) im (n = 19), or 20 mg of dexamethasone plus 5 mg of pRelaxin (n = 17). Length of gestation (in days) was less (P < 0.05) in heifers treated with dexamethasone (279.8 ± 1.0) than in controls (286.6 ± 0.9), but was not influenced (P > 0.05) by treatment with pRelaxin. The incidence of retained placentas in heifers treated only with dexamethasone (27.3%) was not reduced by concomitant treatment with pRelaxin (35.3%). Retained placentas were not observed in any control heifers and in only one heifer (5.2%) treated solely with pRelaxin. Ease of calving (1 = unassisted, 5 = abnormal presentation) was not influenced by treatment (P > 0.05), even though birth weights (in kilograms) of calves from heifers treated with dexamethasone (36.4 ± 0.8) were less (P < .01) than those of calves from nondexamethasone-treated heifers (39.2 ± 0.8). Dexamethasone tended to reduce (P < 0.07) calf vigor (1 = healthy and strong, 5 = dead on arrival; 1.48 ± 0.11 vs. 1.18 ± 0.11), but was not (P > 0.05) influenced by pRelaxin. The duration of the postpartum anestrous interval (73.1 ± 1.8 d across groups) and pelvic areas following treatment and parturition were not influenced (P > 0.05) by dexamethasone or pRelaxin. Although determinants of colostrum quality (P < 0.01) and quantity (P < 0.08) of milk produced were influenced by dexamethasone, adjusted 205-d weights of calves did not differ (P > 0.05) among groups. In conclusion, treatment with pRelaxin alone failed to induce parturition or, when combined with dexamethasone, to reduce the incidence of retained placentas.