Effect of the Medium Replacement Interval on the Viability,Growth and In Vitro Maturation of Isolated Caprine and Ovine Pre‐Antral Follicles

The aim of the present study was to evaluate the effects of different medium replacement intervals on the viability, antral cavity formation, growth and in vitro maturation (IVM) of oocytes from caprine and ovine pre‐antral follicles. Pre‐antral ovarian follicles (≥150 μm) were isolated from the ovarian cortex of goats and sheep and were individually cultured for 24 days using two different medium replacement intervals [2 days (T₁) or 6 days (T₂)]. Follicle development was evaluated on the basis of antral cavity formation, increases in follicular diameter and the presence of healthy cumulus oocyte complexes and fully grown oocytes. For caprine species, results showed a higher percentage (p < 0.05) of viable follicles in T₁ than T₂ from day 6 until the end of the culture. In addition, when comparing both treatments after the same culture duration, the rate of antrum formation was significantly higher in T₁ than in T₂ from day 12 onwards. Yet, in ovines, when both treatments were compared on day 24 of the culture, there were more viable follicles in T₂ than in T₁ (p < 0.05). In the caprine species, percentages of fully grown oocytes (≥110 μm) acceptable for IVM after 24 days of culture were significantly higher in normal follicles cultured in T₁ (30.0%) than in T₂ (6.7%; p < 0.05). On the other hand, in ovines, at the end of the culture, the percentage of oocytes destined for IVM was higher in T₂ than in T₁ (23.5% vs 2.9%; p < 0.05). In conclusion, under the same conditions, the frequency of medium replacement significantly affected the in vitro development of caprine and ovine pre‐antral follicles. To improve the efficiency of the culture system, the medium must be replaced every 2 and 6 days for goat and sheep pre‐antral follicles, respectively.     

Antioxidant, icthyotoxicity and brine shrimp lethality tests of Magonia glabrata

The ethanolic extract of the fruit bark from Magonia glabrata yielded shikimic acid, scopoletin, sitosterol glycoside and 2-O-methyl-l-inositol. Antioxidant, icthyotoxicity and brine shrimp lethality activities were observed in this extract. The major constituent, 2-O-methyl-l-inositol, was found to be inactive in two assays but showed moderate activity as a radical scavenger.     

The influence of the ethanol pumping of wheat straw and of the beating of pulp on the resulting paper sheets

This paper reports on the influence of independent variables in the pulping of wheat straw (viz. cooking temperature and time, and ethanol concentration), and of the number of PFI beating revolutions to which the pulp is subjected, on the yield and Shopper-Riegler index of the pulp, and on the breaking length, stretch, burst index and tear index of the resulting paper sheets. By using a central composite factor design and the BMDP software suite, equations that relate each dependent variable to the different independent variables were obtained that reproduce the experimental results for the dependent variables with errors less than 25% at temperatures, times, ethanol concentrations and numbers of PFI beating revolutions in the ranges 140–180°C, 60–120 min, 40–80% and 0–1750, respectively. Ensuring the optimal Shopper-Riegler index for the pulp (73.18°SR), and also the optimal breaking length (6276 m), stretch (2.57%) and burst index (3.72 kN/g) for the papers sheets, entails using a high temperature (180°C), a large number of PFI beating revolutions (1750) and medium-to-high ethanol concentration and time values; these conditions, however, decrease the tear index for the paper sheets below the optimum level (4.60 mNm²/g). A high temperature and a large number of PFI beating revolutions, in conjunction with a low ethanol concentration (60%) and a short time (60 min), can be used to save solvent and increase the production; the values thus obtained for the dependent variables differ by less than 30% from their optimum levels in the worst case.     

A new dihydroxysterol from the marine phytoplankton Diacronema sp

Diacronema sp. was cultured and its sterols were separated by column chromatography on silica gel. The new sterol 24-ethyl-4alpha-methyl-cholestane-3,20-diol (1) was characterised by NMR and MS spectrometry, as well as (22E)-24-ethyl-4alpha-methyl-5alpha-cholest-22-en-3beta-ol (2) and beta-sitosterol, the major components of the sterol fractions. Neither the biosynthetic origin of the new dihydroxysterol nor its role in the biochemistry of Diacronema is known.     

Linalool production from the leaves of Bursera aloexylon and its antimicrobial activity

The oil of the leaves of Bursera aloexylon was found to contain a high linalool level (96.7 %). The antimicrobial activity tests indicated that the oil was effective against Rhodococcus equi (0.60 mg/ml) and Staphylococcus epidermides (0.15 mg/ml).     

Drying and wetting of Mediterranean soils stimulates decomposition and carbon dioxide emission: the "Birch effect"

Observations on the net carbon exchange of forests in the European Mediterranean region, measured recently by the eddy covariance method, have revived interest in a phenomenon first characterized on agricultural and forest soils in East Africa in the 1950s and 1960s by H. F. Birch and now often referred to as the "Birch effect." When soils become dry during summer because of lack of rain, as is common in regions with Mediterranean climate, or are dried in the laboratory in controlled conditions, and are then rewetted by precipitation or irrigation, there is a burst of decomposition, mineralization and release of inorganic nitrogen and CO(2). In forests in Mediterranean climates in southern Europe, this effect has been observed with eddy covariance techniques and soil respiration chambers at the stand and small plot scales, respectively. Following the early work of Birch, laboratory incubations of soils at controlled temperatures and water contents have been used to characterize CO(2) release following the rewetting of dry soils. A simple empirical model based on laboratory incubations demonstrates that the amount of carbon mineralized over one year can be predicted from soil temperature and precipitation regime, provided that carbon lost as CO(2) is taken into account. We show that the amount of carbon returned to the atmosphere following soil rewetting can reduce significantly the annual net carbon gain by Mediterranean forests.     

Seasonal variation of inorganic nitrogen and net mineralization in a salt marsh ecosystem

Inorganic nitrogen pools and net mineralization were estimated in three sites of a Tagus estuary salt marsh in Portugal throughout 1 year. Ammonium (NH₄ ⁺) was the major form of inorganic nitrogen found in the salt marsh soil. Extractable NH₄ ⁺ concentrations showed a marked seasonal pattern with a concentration peak during the hotter months of July/August. The great majority (>99%) of the total nitrogen in the soil was found in sedimented organic matter, not readily available for plant uptake. Net nitrogen mineralization, determined using a field incubation method, showed a peak during the months of June/July which resulted in an increase on nitrogen availability. With the exception of the lower salt marsh, estimated rates of in situ net nitrogen mineralization in the soil during summer were well related to the increase in plant aboveground biomass and plant nitrogen pools, indicating that the process is an important source of available nitrogen for plant uptake and growth. Annual net nitrogen mineralization ranged between 2.4 and 4.5gNm^–2yr^{– 1} being significantly higher for the lower salt marsh site. Rates of net nitrogen mineralization were relatively low during most of the year with a particularly active period from June to August, possibly due to an effect of temperature on soil microbial activity.     

Variations in growth, survival and carbon isotope composition (delta(13)C) among Pinus pinaster populations of different geographic origins

To evaluate differences in growth and adaptability of maritime pine (Pinus pinaster Ait.), we studied growth, polycyclism, needle tissue carbon isotope composition (delta(13)C) as an estimate of water-use efficiency (WUE) and survival of seven populations at 10 years of age growing in a performance trial at a provenance test site in Escaroupim, Portugal. Six populations were from relatively high rainfall sites in Portugal and southwestern France (Atlantic group), and one population was from a more arid Mediterranean site in Spain. There were significant differences between some populations in total height, diameter at breast height, delta(13)C of bulk needle tissue, polycyclism and survival. A population from central Portugal (Leiria, on the Atlantic coast) was the tallest and had the lowest delta(13)C. Overall, the variation in delta(13)C was better explained by the mean minimum temperatures of the coldest month than by annual precipitation at the place of origin. Analyses of the relationships between delta(13)C and growth or survival revealed a distinct pattern for the Mediterranean population, with low delta(13)C (and WUE) associated with the lowest growth potential and reduced survival. There were significant negative correlations between delta(13)C and height or survival in the Atlantic group. Variation in polycyclism was correlated with annual precipitation at the place of origin. Some Atlantic populations maintained a high growth potential while experiencing moderate water stress. A detailed knowledge of the relationships between growth, survival and delta(13)C in contrasting environments will enhance our ability to select populations for forestry or conservation.     

A new lupene triterpenetriol and anticholinesterase activity of Salvia sclareoides

A new lupene triterpenetriol was isolated from the acetone extract of the aerial parts of Salvia sclareoides and characterised as (1beta,3beta)-lup-20(29)-ene-1,3,30-triol (1). In addition, nepetidin (2), nepeticin (3), lupendiol (4), (1beta,11alpha)-dihydroxy-lup-20(29)-en-3-one (5), ursolic acid (6), sumaresinolic acid (7) and hederagenin (8), were identified in this Salvia sp. To the best of our knowledge, the compounds 2 and 7 are new constituents in Salvia spp. The acetone, ethanol, butanol and water extracts of the plant were screened for the in vitro inhibitory activity of acetylcholinesterase (AChE) and butyrilcholinesterase (BChE), enzymes which play a role in the Alzheimer disease. All extracts inhibited acetylcholinesterase activity at 10 microg/ml, a remarkable activity since the standard drug rivastigmine does not inhibit acetylcholinesterase at the same concentration. Regarding the butyrilcholinesterase, the acetone extract at 1000 microg/ml was able to inhibit completely the enzyme activity and the butanol and ethanol extracts, at this concentration, produced a potent inhibition of BchE.