PCR-RFLP analysis of mitochondrial DNA: a reliable method for species identification

A method for identification of game species has been developed on the basis of the amplification of a specific part of the mitochondrial genome (tRNA(Glu)/cytochrome b) using the polymerase chain reaction (PCR). To distinguish between several game species, the obtained 464-bp-long PCR products were cut with different restriction endonucleases (RE) resulting in species-specific restriction fragment length polymorphism (RFLP). Even closely related deer species could be distinguished by application of one or two RE. Natural polymorphisms of the target sequence within one species were examined for red deer (Cervus elaphus), and base pair substitutions were identified affecting the RFLP pattern.     

Interaction of beta-lactoglobulin with small hydrophobic ligands as monitored by fluorometry and equilibrium dialysis: nonlinear quenching effects related to protein--protein association

Although a thorough characterization of binding parameters is essential for application of beta-lactoglobulin as a carrier for a variety of small hydrophobic ligands, the binding parameters derived in various studies using various techniques are inconsistent. The bindings of several small ligands as detected by fluorometry and equilibrium dialysis were compared. Fluorescence spectroscopy showed that beta-ionone, retinol, and fatty acid lactones all bound in the vicinity of a tryptophan residue. Retinol and fatty acid lactone competed for the same binding site. Exclusively for ligands that quench the beta-lactoglobulin fluorescence through a resonance energy transfer mechanism, fluorometry yielded a systematically higher binding affinity than equilibrium dialysis. The binding overestimation in fluorometric measurements can be explained by oligomer formation of protein, together with an underestimation of the limiting quenching level at saturating ligand concentrations due to the use of a limited set of data points.     

Effects of excessive or restricted phosphorus and calcium intake during early life on markers of bone architecture and composition in pigs

Sufficient supply of pigs with calcium (Ca) and phosphorus (P) is essential for animal health and welfare during the growth period. However, the P content in animal manure is considered as a cause of massive environmental problems in soil and aquatic ecosystems. To complement previous findings, the objective of this study is the investigation of effects of a reduced and increased Ca and P supplementation on bone mineralization and bone structure compared with the current dietary recommendation. Another aim is to find possible serum markers that would allow the assessment of adequacy of P supply for bone health during growth. The result validated that the recommended Ca and P supply is sufficient, without the addition of microbial phytases. However, addition of P has no further beneficial effects on bone stability, while P supplementation below the recommended level affects bone development and growth performance. Reduced P levels have consequences for cancellous bone density and trabecular architecture. Further fine-tuning of the P supply in conjunction with an appropriate Ca supply will contribute to a reduction in P waste and associated environmental impact while maintaining animal health and welfare.     

低温在诱导油桃芽解除休眠过程中对代谢的影响

果树温室栽培时常出现萌芽率低，萌芽开花延迟，花器官发育差，座果率低等问题。调查了分批移入人工气候室的“NJ72”油桃萌芽率，以确定其休眠解除时间，试验结果表明“NJ72”油桃于1月中旬结束休眠，不同温度处理对“NJ72”油桃休眠期代谢的影响不同，低温促进花芽内H2O2的积累，同时促进过氧化氢酶（CAT）、超氧物歧化酶（SOD）、过氧化物酶（POD）活性的提高，低温处理使花芽呼吸速率增加，其中磷酸戊糖途径（PPP）代谢增加，糖酵解（EMP）降低，而三羧酸循环（TCA）代谢波动较小，葡萄糖－6－磷酸（G－6－PDH）脱氢酶活性也有所增加。     

Contribution of nitrification and denitrification to the emission of N2O in a freeze‐thaw event in an agricultural soil

The amounts of N₂O released in freeze‐thaw events depend on site and freezing conditions and contribute considerably to the annual N₂O emissions. However, quantitative information on the N transformation rates in freeze‐thaw events is scarce. Our objectives were (1) to quantify gross nitrification in a Luvisol during a freeze‐thaw event, (2) to analyze the dynamics of the emissions of N₂O and N₂, (3) to quantify the contribution of nitrification and denitrification to the emission of N₂O, and (4) to determine whether the length of freezing and of thawing affects the C availability for the denitrification. ¹⁵NO was added to undisturbed soil columns, and the columns were subjected to 7 d of freezing and 5 d of thawing. N₂O emissions were determined in 3 h intervals, and the concentrations of ¹⁵N₂O and ¹⁵N₂ were determined at different times during thawing. During the 12 d experiment, 5.67 mg NO ‐N (kg soil)^–1 was produced, and 2.67 mg NO ‐N (kg soil)^–1 was lost. By assuming as a first approximation that production and loss occurred exclusively during thawing, the average nitrate‐production rate, denitrification rate, and immobilization rate were 1.13, 0.05, and 0.48 mg NO ‐N (kg soil)^–1 d^–1, respectively. Immediately after the beginning of the thawing, denitrification contributed by 83% to the N₂O production. The ratios of ¹⁵N₂ to ¹⁵N₂O during thawing were narrow and ranged from 1.5 to 0.6. For objective (4), homogenized soil samples were incubated under anaerobic conditions after different periods of freezing and thawing. The different periods did not affect the amounts of N₂ and N₂O produced in the incubation experiments. Further, addition of labile substrates gave either increases in the amounts of N₂O and N₂ produced or no changes which suggested that changes in nutrient availability due to freezing and thawing are only small.     

Resistance of soybean vegetative storage proteins (S-VSPs) to proteolysis by rumen microorganisms

Soybean vegetative storage proteins (S-VSPs) are lysine-rich and, hence, are potentially of high nutritive value for high productive ruminants. Using S-VSPs from wild-type soybean and from transgenic tobacco plants expressing either one of the two S-VSPs subunits (S-VSP alpha or S-VSP beta) or both, we tested their stability in cow rumen fluid under in situ conditions, using SDS-polyacrylamide gel electrophoresis. Proteolysis and degradation pattern of S-VSPs from transgenic tobacco leaves occurred relatively fast compared with that of wild-type (WT) soybean plants. Comparing the two S-VSPs subunits expressed in transgenic plants, we found that S-VSP alpha was degraded much faster than S-VSP beta. The degradation pattern of S-VSPs in transgenic tobacco plants expressing both subunits resembled that of WT soybean. In contrast, the degradation pattern of transgenic tobacco plants expressing a single subunit was different. These finding suggest that the quaternary structure of S-VSPs may be an important factor determining their resistance to rumen degradation. Our results also suggest that the stability to rumen proteolysis of a given protein, when expressed in a transgenic plant, may not always be predictable and has to be verified.     

Glycemic response to a food starch esterified by 1-octenyl succinic anhydride in humans

To evaluate the glycemic response to a food starch esterified by 1-octenyl succinic anhydride (OSA), 30 healthy nondiabetic adult subjects were studied in a double-blind crossover design. After an overnight fast, subjects consumed a product containing either 25 g of glucose or 25 g of OSA-substituted starch. Finger-prick capillary blood was obtained at baseline and 15, 30, 45, 60, 90, and 120 min postprandial for glucose measurement. After OSA treatment, the rise in blood glucose was reduced (P < 0.05) at 15 and 30 min and tended (P < 0.08) to be lower at 45 min. Mean peak rise in glucose was reduced 19% (P < 0.01) by OSA (3.30 +/- 0.19 versus 2.66 +/- 0.16 mmol/L) compared to glucose, but time to peak did not differ between treatments. Net incremental area under the curve was also lower (P < 0.05) on OSA compared to glucose. Minimal effects on gastrointestinal symptoms (intensity and frequency of nausea, cramping, distention, and flatulence) were noted for both products, with no clinically significant difference between products. In conclusion, starch substitution with OSA attenuated the postprandial glycemic excursion compared to an equivalent glucose challenge and was well tolerated by fasting healthy adult subjects.     

Determining the rate of change in a mixed deciduous forest monitored for 50 years

Introduction   

Trees in two compartments of the mixed deciduous forest Draved Forest have been monitored regularly for 50 years.