BACKGROUND: Endogenous lysine in ileal digesta has been determined directly using highly digestible proteins and hydrolysates and assuming a complete absorption of dietary amino acids. In this study, the endogenous lysine in the ileal digesta of rats fed casein was determined using the guanidination method (baseline) and directly (assuming complete digestion and absorption) for casein and a highly hydrolysed casein. The peptide alimentation/ultrafiltration technique was also used to determine the endogenous ileal lysine for the casein hydrolysate (Peptopro®).RESULTS: Mean endogenous ileal lysine for rats given the unguanidinated casein and determined directly assuming complete digestion and absorption (644 mg kg⁻¹ dry matter intake (DMI)) or for a highly hydrolysed casein with the same assumption (596 mg kg⁻¹ DMI) was significantly (P < 0.05) higher than the endogenous ileal lysine for casein determined using the guanidination method (274 mg kg⁻¹ DMI). The endogenous lysine in ileal digesta for a casein hydrolysate determined using the alimentation/ultrafiltration method (302 mg kg⁻¹ DMI) was not significantly (P < 0.05) different from that based on the guanidination method but was significantly (P < 0.05) lower than the equivalent endogenous lysine determined directly.CONCLUSION: Dietary amino acids are not completely absorbed from casein or its hydrolysate and determining endogenous amino acids in ileal digesta directly, assuming complete absorption, overestimates the amount of endogenous ileal amino acids.

The genetic variation within and between 13 populations (385 individuals) of Pinus uncinata was analyzed with ten chloroplast microsatellite markers. Both the infinite allele mutation and stepwise mutation model (SMM) have been applied to the analysis of the genetic structure and the geographical distribution of haplotypic variation. High level of genetic diversity and low but significant differentiation among compared population were found. Three marginal populations, Sierra de Cebollera, Margaride Mountains and Sierra de Gúdar are strongly differentiated from the rest. Mutations following SMM-like process contributed significantly to the regional differentiation. The pattern of genetic structure observed in mountain pine is common in conifers with a wide distribution range. Lack of significant genetic structuring may be a result of a recent fragmentation of a historically larger population and/or interspecific hybridization and introgression. The southernmost populations from the Sierra Cebollera and the Sierra de Gúdar are the most genetically distinct. This suggests a long period of spatial isolation and/or origin from different ancestral populations.

In this study, a biomonitoring project using the moss Scorpiurum circinatum was carried out to evaluate the deposition and biological effects of heavy metals in the area of Acerra (Naples, S Italy), one of the vertices of the sadly called "Italian triangle of death" owing to the dramatic increase in tumours. The results clearly indicated that the study area is heavily polluted by heavy metals, a large proportion of which is likely present in the atmosphere in particulate form. The ultrastructural organization of exposed samples was essentially preserved, but cell membrane pits, cytoplasm vesicles and concentric multilamellar/multivesicular bodies, probably induced by pollution, were found, which may be involved in the tolerance mechanisms to metal pollution in this moss species. Although severe biological effects were not found at the ultrastructural level in the exposed moss, effects on humans, especially after long-term exposure, are to be expected. The moss Scorpiurum circinatum indicates that the "Italian triangle of death" is heavily polluted by heavy metals.

Two calcified structures, the eggshell and sperm-associated body (SB), are present in the eggs of the Japanese quail, Coturnix japonica. X-ray diffractometry showed that calcium carbonates take the form of calcite in the eggshell and aragonite in the SB. The aim of the present study was to identify the factors that determine the morphology of calcium carbonate crystals. The matrix of EDTA-treated eggshell was a meshwork of vesicles, 200 to 500 nm in diameter, connected by fine fibers or fibrous sheets. The matrix of SB cortex was a radiation of rod-shaped projections approximately 130 nm in width. In vitro crystal formation was achieved by adding dissociated matrix substances to test solutions. When eggshell matrix material was added, formation of calcite crystals, which had many vesicular holes on their surface, was observed. When SB matrix material dissociated by sonication was added, rhombohedral calcite crystals formed at protein concentrations of 100 μg/mL or lower, and elongated and bundled crystals formed at concentrations of 150 μg/mL or higher. When SB matrix material dissociated by pipetting was added, aragonite crystals formed. These observations indicate that the matrix structure is the principal factor in determining the crystal polymorphism of calcium carbonate.

Transgenic loci obtained after Agrobacterium tumefaciens-mediated transformation can be simple, but fairly often they contain multiple T-DNA copies integrated into the plant genome. To understand the origin of complex T-DNA loci, floral-dip and root transformation experiments were carried out in Arabidopsis thaliana with mixtures of A. tumefaciens strains, each harboring one or two different T-DNA vectors. Upon floral-dip transformation, 6-30% of the transformants were co-transformed by multiple T-DNAs originating from different bacteria and 20-36% by different T-DNAs from one strain. However, these co-transformation frequencies were too low to explain the presence of on average 4-6 T-DNA copies in these transformants, suggesting that, upon floral-dip transformation, T-DNA replication frequently occurs before or during integration after the transfer of single T-DNA copies. Upon root transformation, the co-transformation frequencies of T-DNAs originating from different bacteria were similar or slightly higher (between 10 and 60%) than those obtained after floral-dip transformation, whereas the co-transformation frequencies of different T-DNAs from one strain were comparable (24-31%). Root transformants generally harbor only one to three T-DNA copies, and thus co-transformation of different T-DNAs can explain the T-DNA copy number in many transformants, but T-DNA replication is postulated to occur in most multicopy root transformants. In conclusion, the comparable co-transformation frequencies and differences in complexity of the T-DNA loci after floral-dip and root transformations indicate that the T-DNA copy number is highly determined by the transformation-competent target cells.

1.The atolls and islands comprising Chagos are a biodiversity hotspot of global conservation significance in a remote part of the central Indian Ocean.2.This study examines the condition of the archipelago's coastal ecosystems by rapid environmental assessment at 21 sites/islands, which were also investigated a decade earlier using the same methodology. Major changes in ecosystem structure and environmental disturbance were determined.3.Coral fish abundance was significantly lower in 2006 than 1996. Decrease in the physical structural complexity of the reefs, as a result of coral bleaching and mortality induced by the 1998 warming event, may have been a contributing factor.4.Evidence of collecting/fishing was significantly greater in 2006 than 1996. This is attributed mainly to an illegal fishery for holothurians (sea cucumbers), which has expanded over recent years and now exerts substantial pressure on the resource. The significant decline observed in beach wood, a readily accessible fuel for fishing camps, is consistent with this.5.Solid waste on islands was high (median 2 to 20 items m⁻¹ beach) in both 1996 and 2006. Potentially harmful biological impacts, determined from other studies, include entanglement, toxic effects and provision of transport for invasives or other 'hitchhiker' species.6.Significantly higher bird abundances were recorded in protected areas than 'unprotected' areas, attributed mainly to absence of predation by rats.7.Rapid assessment augments more comprehensive ecosystem investigations. It provides a valuable snapshot of environmental conditions based upon a broad suite of features (ecosystems and disturbances) determined, concurrently, within the same site inspection quadrats and using the same scale of assessment.

Among the many toxic metals, the biokinetics of copper (Cu) in the freshwater cladoceran Daphnia magna have not been studied due to the lack of an ideal radiotracer. In the present study, a gamma radiotracer, 67Cu (half-life = 61.9 h), was used to study the uptake of copper from the dissolved and dietary phase and efflux in D. magna, an important toxicity testing species. The influx rate of Cu from the dissolved phase increased with dissolved Cu concentration, with a calculated uptake rate constant of 0.055 L/g/h. The assimilation efficiency (AE) of Cu decreased significantly (from 92 to 16%) as the available food concentration increased, and the AE differed among the food types. As low as 1% of Cu AE was found in daphnids fed high concentrations (1.54 mg/L) of the green algae Chlorella pyrenoidosa. The AE decreased linearly as the ingestion rate of the daphnids increased. The efflux rate constant was 0.20/d at high food concentrations. Excretion accounted for 82 to 94% of total Cu loss from the animals, although Cu also was transferred maternally from female adults to their offspring. Under conditions of high food concentrations, approximately 6.5% of the mother's Cu was transferred to the offspring over 7 d. It was concluded that Cu accumulation is dominated by uptake from dietary sources, and there is a substantial need to understand the dietary toxicity of Cu to daphnids. The present study has implications for the choice of food particles in conducting the Cu toxicity testing in cladocerans.

In this biological oxygen demand (BOD) study, the manometric respirometric BOD OxiTop® method was used to monitor the biodegradation of two summer grade (SFO 1 and 2) and two winter grade light fuel oils (WFO 1 and 2) in OECD 301 F conditions, in groundwater, and in two different Finnish forest soils (mineral-poor and mineral-rich). The biodegradation measurements in the OECD 301 F conditions were carried out in two nutrient solutions for 28 days. In both solutions WFO 1 reached the highest biodegradation degree, 32% in the solution OECD 301 F, and 70% in a solution containing additional ammonium chloride. In groundwater conditions all the biodegradation degrees of fuel oils remained below 2% within the 28-day period. SFO 1 reached the highest 30 day biodegradability (4%) in mineral-poor soil, 18% in mineral-rich soil. In a 189-day measurement in a mineral-rich soil, the biodegradation degree for the SFO 1 was 94%. The manometric respirometric method proved to be a very suitable and practicable measurement method for the purpose of biodegradation studies of highly volatile light fuel oils, because in this method samples are treated to a lesser degree than in conventional methods, and dilutions are not needed. Results also indicated a considerable effect of conditions on the biodegradability in both water and soil environments. The results of these biodegradation studies could be used when planning in situ treatment methods based on natural biodegradation. In situ treatment methods are eco-efficient, and are especially suitable for sparsely populated sites.

In this study, a modified version of the gas production technique was used to determine protein fermentation characteristics in rumen fluid of 19 feedstuffs. Performing the incubations in a N-free environment, and with an excess of rapidly fermentable carbohydrates, made N the limiting factor to microbial growth, and so gas production profiles reflected the availability of N from the feed samples. Results showed that fermentation of protein in rumen fluid can be determined with this modified gas production technique, and that there were distinct differences in protein fermentation between the feed samples. Availability of protein for fermentation was highest in wheat, potato pieces and lupin, and lowest in Rumiraap, a formaldehyde treated rapeseed meal, palm kernel expeller and brewery grains. The protein degradation characteristics of the 19 feed ingredients were also determined with the in situ nylon bag technique. With the obtained results, the amount of rumen escape protein (REP) was calculated for each feedstuff. The results showed that the rate of degradation ranged from 0.010/h for Rumiraap to 0.151/h for wheat. The amount of REP ranged from 197g/kg CP for lupin to 840g/kg CP for Rumiraap. Comparing the gas production results with the results obtained with the nylon bag technique showed that there was a good relationship between the gas production after 12-25h of incubation and the calculated amount of REP (r ² =0.83-0.85). The results show that the adapted gas production technique, being depleted of N and using an excess of rapidly fermentable carbohydrates, is suitable to recognize differences in N availability between feed samples and can be used as an alternative to the nylon bag technique and other in vitro techniques.

consultable en ligne | International audience | Sturgeon disappeared from the Rhône River in the mid 70's without certitude about which species it was and about the existence of a sympatry between European sturgeon, Acipenser sturio, and Adriatic sturgeon, A. naccarii, in this watershed. In order to reach a reliable specific determination of this extinct sturgeon population, archaeozoological remains of the Jardin d'Hiver in Arles city, on the Rhône river banks, were genetically analysed, following strict criteria of authentication for the ancient DNA work. The rich collection of Arles sturgeon bone remains stems from human activities between the 6th and the 2nd Century BC. Sequences of 86 bp of the cytochrome b gene were obtained on four bones, from different anatomical parts of the fish and from different archaeological layers. All gave A. sturio diagnostic sequences. This preliminary analysis is an essential first step in the project of sturgeon reintroduction in the Rhône River. Thus, further analyses on a larger sample are necessary to comfort this result and to solve the question of sympatry with A. naccarii.