Developmental competence of heat stressed oocytes from Holstein and Limousine cows matured in vitro

The negative effects of heat stress on dairy cattle's fertility have been extensively studied, but the relevant knowledge for beef cattle is rather limited. The aims of this study were to investigate the effects of HS during in vitro maturation on the developmental potential of oocytes derived from Limousine and Holstein cows and to estimate the effect of the differential gene expression of important genes in oocytes, cumulus cells and blastocysts in the growth competence between the breeds. In seven replicates, cumulus oocyte complexes from Holstein and Limousine cows were matured for 24 hr at 39°C (controls C; Hol_39, Lim_39) or at 41°C from hour 2 to hour 8 of IVM (treated T; Hol_41, Lim_41), fertilized, and presumptive zygotes were cultured for 9 days at 39°C. Cleavage and embryo formation rates were evaluated 48 hr post-insemination and on days 7, 8 and 9, respectively. From all groups, subsets of cumulus cells, oocytes and blastocysts were analysed for the relative expression of genes related to metabolism, stress, apoptosis and placentation. No difference was detected in cleavage rate or in blastocyst formation rate among the control groups. In both breeds, heat stress reduced blastocyst yield, but at all days the suppression was higher in Limousines. In Holsteins, altered gene expression was detected in cumulus cells (G6PD, GLUT1) and blastocysts (PLAC8), while in Limousines, differences were found in oocytes (G6PD, HSP90AA1), in cumulus cells (CPT1B, HSP90AA1, SOD2) and blastocysts (DNMT, HSP90AA1, SOD2). It appears that Holstein COCs are more tolerant than Limousine COCs, possibly due to compulsory, production driven selection.     

Ultrasonographic findings of the corpus luteum and the gravid uterus during heat stress in dairy cattle

The objectives of this study were to assess alterations in, echogenic appearance, size and blood flow in the corpus luteum, the placentomes and the blood flow in umbilical and uterine arteries that heat stress can cause in cooled pregnant dairy cows. Pregnant cows were allocated in two groups and the gravid uteri, along with the ipsilateral corpora lutea were examined during the winter (group W, n = 9) or the summer (group S, n = 10). The grey-scale ultrasound and colour flow imaging of the corpus luteum and placentome were performed. In addition, the umbilical and uterine artery diameters and haemodynamic parameters in the vessels were calculated. At the time of ultrasonographic examination, cortisol concentrations were higher, and progesterone levels tended to be lower in group S compared to group W. The grey-scale ultrasound evaluation of corpora lutea and placentomes was lower in group S compared to group W. The diameter of umbilical artery and the blood volume in the vessel were less in group S than in group W. We infer that heat stress affects foetal blood supply and possibly the structure of placentomes and corpora lutea, but it differently affects the blood flow characteristics in the umbilical and uterine arteries.     

Deficit irrigation based on drought tolerance and root signalling in potatoes and tomatoes

Agriculture is a big consumer of fresh water in competition with other sectors of the society. Within the EU-project SAFIR new water-saving irrigation strategies were developed based on pot, semi-field and field experiments with potatoes (Solanum tuberosum L.), fresh tomatoes (Lycopersicon esculentum Mill.) and processing tomatoes as model plants. From the pot and semi-field experiments an ABA production model was developed for potatoes to optimize the ABA signalling; this was obtained by modelling the optimal level of soil drying for ABA production before re-irrigation in a crop growth model. The field irrigation guidelines were developed under temperate (Denmark), Mediterranean (Greece, Italy) and continental (Serbia, China) climatic conditions during summer. The field investigations on processing tomatoes were undertaken only in the Po valley (North Italy) on fine, textured soil. The investigations from several studies showed that gradual soil drying imposed by deficit irrigation (DI) or partial root zone drying irrigation (PRD) induced hydraulic and chemical signals from the root system resulting in partial stomatal closure, an increase in photosynthetic water use efficiency, and a slight reduction in top vegetative growth. Further PRD increased N-mineralization significantly beyond that from DI, causing a stay-green effect late in the growing season. In field potato and tomato experiments the water-saving irrigation strategies DI and PRD were able to save about 20-30% of the water used in fully irrigated plants. PRD increased marketable yield in potatoes significantly by 15% due to improved tuber size distribution. PRD increased antioxidant content significantly by approximately 10% in both potatoes and fresh tomatoes. Under a high temperature regime, full irrigation (FI) should be undertaken, as was clear from field observations in tomatoes. For tomatoes full irrigation should be undertaken for cooling effects when the night/day average temperature >26.5 °C or when air temperature >40 °C to avoid flower-dropping. The temperature threshold for potatoes is not clear. From three-year field drip irrigation experiments we found that under the establishment phase, both potatoes and tomatoes should be fully irrigated; however, during the later phases deficit irrigation might be applied as outlined below without causing significant yield reduction:

•

Potatoes

°

After the end of tuber initiation, DI or PRD is applied at 70% of FI. During the last 14 days of the growth period, DI or PRD is applied at 50% of FI.

•

Fresh tomatoes

°

From the moment the 1st truce is developed, DI is applied at 85-80% of FI for two weeks. In the middle period, DI or PRD is applied at 70% of FI. During the last 14 days of the growth period, DI or PRD is applied at 50% of FI.

•

Processing tomatoes

°

From transplanting to fruit setting at 4th-5th cluster, the PRD and DI threshold for re-irrigation is when the plant-available soil water content (ASWC) equals 0.7 (soil water potential, Ψ_soil = −90 kPa). During the late fruit development/ripening stage, 10% of red fruits, the threshold for re-irrigation for DI is when ASWC = 0.5 (Ψ_soil = −185 kPa) and for PRD when ASWC (dry side) = 0.4 (Ψ_{soil, dry side} = −270 kPa).

The findings during the SAFIR project might be used as a framework for implementing water-saving deficit irrigation under different local soil and climatic conditions.     

Protective and defensive roles of non-glandular trichomes against multiple stresses:structure-function coordination

As superficial structures,non-glandular trichomes,protect plant organs against multiple biotic and abiotic stresses.The protective and defensive roles of these epidermal appendages are crucial to developing organs and can be attributed to the excellent combination of suitable structural traits and chemical reinforcement in the form of phenolic compounds,primarily fl avonoids.Both the formation of trichomes and the accumulation of phenolics are interrelated at the molecular level.During the early stages of development,non-glandular trichomes show strong morphological similarities to glandular ones such as the balloon-like apical cells with numerous phenolics.At later developmental stages,and during secondary wall thickening,phenolics are transferred to the cell walls of the trichomes.Due to the diff use deposition of phenolics in the cell walls,trichomes provide protection against UV-B radiation by behaving as optical fi lters,screening out wavelengths that could damage sensitive tissues.Protection from strong visible radiation is also aff orded by increased surface light refl ectance.Moreover,the mixtures of trichome phenolics represent a superfi-cial chemical barrier that provides protection against biotic stress factors such as herbivores and pathogens.Although the cells of some trichomes die at maturity,they can modulate their quantitative and qualitative characteristics during development,depending on the prevailing conditions of the external biotic or abiotic environment.In fact,the structure and chemical constituents of trichomes may change due to the particular light regime,herbivore damage,wounding,water stress,salinity and the presence of heavy metals.Hence,trichomes represent dynamic protective structures that may greatly aff ect the outcome of many plant–environment interactions.     

Relationships of Spectral Reflectance with Plant Tissue Mineral Elements of Common Bean (Phaseolus Vulgaris L.) Under Drought and Salinity Stresses

ABSTRACT

Salinity and drought stresses are critical for Phaseolus vulgaris L. growth and development. They affect plants in various ways, including tissue mineral element content. Micro- and macro-elements in leaves of Phaseolus vulgaris L. (cv. ‘Blue lake’ and cv. ‘Terli’) subjected to deficit irrigation and salinity treatments were investigated, both analytically and with regards to their effect on the leaves’ spectral reflectance. B (boron), K (potassium), Mn (manganese), Na (sodium), Si (silicon) and Zn (zinc) appeared to be influenced by stress factors, mainly responding to salinity increase. The leaf spectral reflectance of the plants appeared to be significantly correlated with most of the elements under investigation. Multivariate regression identified a relationship of the reflectance at particular regions of the spectrum with phosphorus and NDVI (normalized difference vegetation index) and indicated a significant correlation with B, Fe (iron), K, Mn, P (phosphorus) and Zn. Moreover, customized spectral indices, exhibiting significantly high correlation with B, Fe, K, Mg (magnesium), Mn, Na, P, Zn and N (nitrogen), were developed.     

Vermiculite in Fluidized Bed as Decontaminating Agent for Liquid Phases

Over the last decades, removal of potentially toxic and hazardous materials has received a great deal of attention in the field of environmental pollution. Problems associated with the disposal of the wastes of different kinds of industries led to studies of the sorption?Cuptake properties of clay minerals and zeolites. In the present research, the behavior of vermiculite particles ranging between 425 and 500???m in a laboratory-scale fluidized bed column for uptake of Cs, Hg, and Mn ions from aqueous solutions and wastes in the presence of competing cations has been studied in order to investigate techniques for decontamination of liquid phases. Vermiculite selectively removed high percentages of Cs even from low concentrations in the presence of competing cations. Also removed were up to 60?% of added Hg²⁺ at concentrations of 5?ppm from drinking water and about 84?% from seawater, and furthermore, Mn²⁺ was selectively removed from low-concentration (ca 10?ppm) industrial wastes even when the ratio of Mn²⁺ to competing cations was 1:94. The results suggest the potential use of vermiculite as decontaminating agent in well-designed fluidized bed columns.     

Determination of anabolic steroids in muscle tissue by liquid chromatography-tandem mass spectrometry

A specific and sensitive method based on liquid chromatography-tandem mass spectrometry using atmospheric pressure chemical ionization (LC-APCI-MS/MS) has been developed for the determination of four anabolic steroids [trenbolone, methylboldenone, methyltestosterone, and norethandrolone] in bovine muscle. Methyltestosterone- d 3 was used as internal standard. The procedure involved enzymatic hydrolysis, extraction with tert-butyl methyl ether, defattening, and final cleanup with solid-phase extraction with Oasis HLB cartridges. The analytes were analyzed by reversed-phase LC-MS/MS, acquiring two diagnostic product ions from the chosen precursor [M + H] (+) for the unambiguous confirmation of hormones. The method was validated according to the European Commission Decision 2002/657/EC for the detection and confirmation of residues in products of animal origin. The limits of detection (LOD) and limits of quantitation (LOQ) were found to be 0.3 ng/g and 1.0 ng/g, respectively. The accuracy and precision have been determined, with recoveries ranging from 83% to 104% and the CV factor not exceeding the value of 7%. The decision limits CCalpha were calculated and ranged from 0.05 to 0.15 ng/g while the detection capabilities CCbeta ranged from 0.09 to 0.25 ng/g. The method proved to be sensitive and reliable and thus renders an appropriate means for residue analysis studies.     

Anti-Inflammatory Potential of Monogalactosyl Diacylglycerols and a Monoacylglycerol from the Edible Brown Seaweed Fucus spiralis Linnaeus

A monoacylglycerol (1) and a 1:1 mixture of two monogalactosyl diacylglycerols (MGDGs) (2 and 3) were isolated from the brown seaweed Fucus spiralis Linnaeus. The structures were elucidated by spectroscopic means (NMR and MS) and by comparison with the literature. Compound 1 was composed of a glycerol moiety linked to oleic acid (C18:1 Ω9). Compounds 2 and 3 contained a glycerol moiety linked to a galactose unit and eicosapentaenoic acid (C20:5 Ω3) combined with octadecatetraenoic acid (C18:4 Ω3) or linolenic acid (C18:3 Ω3), respectively. The isolated compounds were tested for their cytotoxic and anti-inflammatory activity in RAW 264.7 macrophage cells. All of them inhibited NO production at non-cytotoxic concentrations. The fraction consisting of compounds 2 and 3, in a ratio of 1:1, was slightly more effective than compound 1 (IC₅₀ of 60.06 and 65.70 µg/mL, respectively). To our knowledge, this is the first report of these compounds from F. spiralis and on their anti-inflammatory capacity.     

Interactions between residue placement and earthworm ecological strategy affect aggregate turnover and N2O dynamics in agricultural soil

Previous laboratory studies using epigeic and anecic earthworms have shown that earthworm activity can considerably increase nitrous oxide (N₂O) emissions from crop residues in soils. However, the universality of this effect across earthworm functional groups and its underlying mechanisms remain unclear. The aims of this study were (i) to determine whether earthworms with an endogeic strategy also affect N₂O emissions; (ii) to quantify possible interactions with epigeic earthworms; and (iii) to link these effects to earthworm-induced differences in selected soil properties. We initiated a 90-day ¹⁵N-tracer mesocosm study with the endogeic earthworm species Aporrectodea caliginosa (Savigny) and the epigeic species Lumbricus rubellus (Hoffmeister). ¹⁵N-labeled radish (Raphanus sativus cv. Adagio L.) residue was placed on top or incorporated into the loamy (Fluvaquent) soil. When residue was incorporated, only A. caliginosa significantly (p < 0.01) increased cumulative N₂O emissions from 1350 to 2223 μg N₂O-N kg⁻¹ soil, with a corresponding increase in the turnover rate of macroaggregates. When residue was applied on top, L. rubellus significantly (p < 0.001) increased emissions from 524 to 929 μg N₂O-N kg⁻¹, and a significant (p < 0.05) interaction between the two earthworm species increased emissions to 1397 μg N₂O-N kg⁻¹. These effects coincided with an 84% increase in incorporation of residue ¹⁵N into the microaggregate fraction by A. caliginosa (p = 0.003) and an 85% increase in incorporation into the macroaggregate fraction by L. rubellus (p = 0.018). Cumulative CO₂ fluxes were only significantly increased by earthworm activity (from 473.9 to 593.6 mg CO₂-C kg⁻¹ soil; p = 0.037) in the presence of L. rubellus when residue was applied on top. We conclude that earthworm-induced N₂O emissions reflect earthworm feeding strategies: epigeic earthworms can increase N₂O emissions when residue is applied on top; endogeic earthworms when residue is incorporated into the soil by humans (tillage) or by other earthworm species. The effects of residue placement and earthworm addition are accompanied by changes in aggregate and SOM turnover, possibly controlling carbon, nitrogen and oxygen availability and therefore denitrification. Our results contribute to understanding the important but intricate relations between (functional) soil biodiversity and the soil greenhouse gas balance. Further research should focus on elucidating the links between the observed changes in soil aggregation and controls on denitrification, including the microbial community.