The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa

To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor^? probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor^? with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor^? Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor^? labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor^? was compared with SYBR‐14/PI, the agreement was high (mean of differences: ?0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor^? did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor^? might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.     

Prevalence of methicillin-resistant Staphylococci on a farm: staff can harbour MRS when animals do not

The aim of this work was to establish the prevalence of methicillin-resistant Staphylococci (MRS) in the animals and staff of a teaching and research farm. Samples of dairy cattle (36), beef cattle (26), sheep (19), horses (21), pigs (23), goats (23) and humans (13) were collected and screened for the presence of MRS. The detection of mecA gene was performed by PCR to determine the resistance of the samples to methicillin. Antimicrobial-resistance testing to penicillin, meropenem, ceftriaxone, cephalothin, oxacillin, levofloxacin, enrofloxacin, chloramphenicol, ciprofloxacin, gentamicin, clindamycin, erytromycin, linezolid, sulfamethoxazole/trimethoprim, tetracycline, doxycycline and vancomycin was performed on the mecA+ isolates. From the 161 samples, four methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were isolated from human beings (31%), whereas none was isolated from animals (0%). No methicillin-resistant Staphylococcus aureus (MRSA) were isolated. All of the MRCoNS isolates from this work presented different antimicrobial resistance patterns. MRCoNS may be present in humans associated with animals while not present in the animals. Selective pressure outside of the farm and a lack of MRCoNS transmission between humans and animals may be responsible for this lack of correlation.     

Behaviour of bentazon as influenced by water and tillage management in rice‐growing conditions

Bentazon is a widely used herbicide in rice agroecosystems that has commonly been found in water resources. To assess how tillage and water regimes affect sorption/desorption, dissipation and leaching of bentazon in Mediterranean rice‐growing conditions, field experiments were carried out using tillage and flooding (TF), tillage and sprinkler irrigation (TS), no‐tillage and sprinkler irrigation (NTS) and long‐term no‐tillage and sprinkler irrigation (NTS7). After 3 years, the K_d values in TS were 2.3, 1.6 and 1.7 times lower than the values in NTS7, NTS and TF respectively. Greater sorption of bentazon was related to higher contents in total organic carbon and, although to a lesser extent, in humic acids and dissolved organic carbon. The persistence of bentazon was significantly greater under anaerobic (half‐life DT₅₀ = 94.1–135 days) than under aerobic (DT₅₀ = 42.4–91.3 days) incubation conditions for all management regimes. Leaching losses of bentazon were reduced from 78 and 74% in TS and TF to 61 and 62% in NTS7 and NTS respectively. The mid‐ and long‐term implementation of sprinkler irrigation in combination with no‐tillage could be considered a management system that is effective at reducing water contamination by bentazon in Mediterranean rice‐growing agroecosystems. © 2017 Society of Chemical Industry     

Effects of different bronopol treatments on final survival rates in the artificial incubation of crayfish eggs (Pacifastacus leniusculus,Astacidae)

Two experiments were conducted on the effects of bronopol in the artificial incubation of crayfish eggs (Pacifastacus leniusculus) with the aim to search an alternative to formaldehyde. In the first experiment, 50, 250, 500 and 1000 ppm bronopol and 3000 ppm formaldehyde (control) in periodical administrations were tested on a density of 6.6 eggs cm^?2. After 44 days of incubation, the highest survival was obtained with 1000 ppm bronopol (81.9% to stage 2 juvenile, with no significant difference from formaldehyde), whereas lower bronopol concentrations resulted in significantly lower survival. In the second experiment, 1000, 3000 and 5000 ppm bronopol and 3000 ppm formaldehyde (control) administered for 15 min every second day were tested on eggs at a density of 20 eggs cm^?2. After 78 days of incubation, bronopol at 3000 ppm allowed for a stage 2 juvenile survival rate of 65.0% (with no significant difference from formaldehyde), whereas significantly lower survival was obtained with 1000 ppm or 5000 ppm. This study shows that bronopol may constitute an alternative to formaldehyde in the artificial incubation of crayfish eggs. A concentration of 3000 ppm administered for 15 min every second day may be adequate even on long incubations at high densities (at least 20 eggs cm^?2, one complete layer).     

Regulated deficit irrigation in ‘Clementina de Nules’ citrus trees. II: Vegetative growth

Summary

An experiment on Regulated Deficit Irrigation (RDI) was performed during 1995 and 1996 in an orchard planted with drip-irrigated ‘Clementina de Nules’/Carrizo Citrange in Moncada (Valencia) Spain. Treatments consisted of a control, irrigated during the whole year at 125% ET_lys and RDI treatments where irrigation was reduced to 25% or to 50% of crop evapotranspiration measured by a weighing lysimeter (ET_lys) during one of the following periods: I) flowering and fruit set (spring); II) initial fruit enlargement phase (summer) and III) final fruit growth and maturation phases (end of summer-autumn). An additional treatment, denominated 50%-Year, was irrigated at 50% ET_lys during the whole year. The effects of RDI treatments in relation to tree water status (pre-dawn Ψ^pd and midday Ψ^md leaf water potential, as well as their integral with time) show a good relation between total shoot emergence in the different growth flushes and the stress intensity reached (Ψ^pd) (r² = 0.80). This correlation was mainly due to the number of floral shoots (r² = 0.86) and not to vegetative ones (r² = 0.22). Similar results were observed between the stress integral at pre-dawn in each period and the former sprouting variables. In all cases, correlation was better with pre-dawn leaf water potential or with pre-dawn stress integral than with those at midday. RDI during spring reduced shoot length of the first growth flush (A1) and increased fruitlet fall after restarting normal irrigation. It also produced “off-season” flowering in the second flush growth (A2) and increased shoot emergence of the third flush growth (A3) with about 10% of them being floral. Summer RDI treatments did not alter vegetative growth, and although they produced off-season flowering (A3) it was much smaller than that of autumn RDI treatments, which in addition reduced vegetative growth with respect to the control. These effects, together with those of yield and fruit quality presented elsewhere, show that summer is the more appropriate period to apply RDI in “Clementina de Nules” mandarin trees.     

Absorptive root areas of large pedunculate oak trees differing in health status along a road in South Bohemia,Czech Republic

The health status of 200- to 400-year-old trees growing in an area alongside a road was described in detail on the basis of evaluation of hazard trees in urban areas and analyzed using the acoustic scanning and modified earth impedance (MEI) methods. Acoustic analysis was used to characterize tree trunks and coarse roots; MEI was used to characterize the active absorptive root surfaces (the electric impedance of soils and sapwood were also included). Several groups of trees were distinguished according to the extent of damage (slight, serious or extreme), which was easily detectable from the MEI results. It was confirmed that both methods gave similar results, which also corresponds to the detailed observations. Instrumental measurements provided additional information about trunk and root injuries, which cannot be directly observed. A combination of both methods is recommended to achieve the most reliable results. This unique approach is applicable for testing eventual threats by old ill trees for safety reasons over the landscape.     

Fluorine immission to acid soil in the vicinity of an aluminium smelter in Galicia (NW Spain) and its influence on aluminium dynamics

Purpose

The aluminium smelter located in the northern coast of Galicia (NW Spain) is a source of fluoride pollution. Previous papers Gago et al. (Commun Soil Sci Plant Anal 32:2503–2517, 2001, Fluoride 35:110–121, 2002) have identified an area of 3-km radius around the factory being affected by fluoride emissions. This study aims to investigate the influence of fluoride emissions on fluorine concentration and speciation as well as on aluminium speciation in soils in the vicinity of the smelter.

Materials and methods

PVC cylinders containing soil material from the A horizon of an unpolluted forest soil, similar to that in the vicinity of the factory, were placed within the A horizon of soils surrounding the smelter. The cylinders were placed at various distances (up to 3 km) from the factory in the direction of predominant winds. The PVC columns were removed after 6 months, and the soil material inside was divided into two slices (0–10 and 10–20 cm) and analysed for pH, total and available fluorine, exchangeable cations, effective cation exchange capacity (CEC) and oxalate-extracted aluminium. The soil solution was mimicked by aqueous extracts and analysed for pH, electrical conductivity, Ca, Mg, Na, K, total and free fluorine, total Al, acid-soluble Al, non-labile Al, labile Al and labile Al species.

Results and discussion

In the area closest to the factory (0.5 km), total and available fluorine in the upmost 10 cm soil increased by twofold and eightfold, respectively, while soluble fluoride increased by fourfold. Increases were also observed in exchangeable Ca, Mg, Al and CEC. Noncrystalline Al increased significantly at any distance from the smelter. Fluorine, pH, sulphate, aluminium and organic matter increased in the soil solution of surface (<10 cm) samples closest to the factory. Monomeric Al-organic matter complexes prevailed in the liquid phase at 0.5–1 km from the factory whereas inorganic Al complexes did at distances over 1 km. Among the inorganic Al complexes, fluoride complexes prevailed at distances less than 1 km and hydroxylated complexes at distances greater than 1 km. Al³⁺ concentration was always very low.

Conclusions

Emissions from an aluminium smelter led to increased concentrations of fluorine and various forms of aluminium in the nearby soil. These emissions mobilised aluminium, organic matter and probably sulphate from the soil. In the closest neighbourhood of the smelter, it was found that most soluble aluminium was bound to organic matter, hence having low toxicity, while at distances farther than 1 km Al-F and Al-OH complexes prevailed.     

Physico‐chemical characterization and in vitro digestibility of commercial feeds used in weaning of marine fish

Six different commercial feeds used in weaning and postweaning of marine fish were analysed from the perspective of some of their main physico‐chemical features and in vitro digestibility. While the range of particle sizes in dry state closely agreed to those claimed by the producing companies, dramatic changes were observed after suspension in water. Sedimentation rates measured in some of the feeds with higher particle sizes may be too high to ensure intake by young fish. The label claims of the products overestimate the amount of total protein but underestimate that of lipids. The analysis of amino acid profiles evidenced important differences in total amino acid composition between feeds of the same company but a great similitude in the profile of free amino acids (FAA). The evaluation of leaching evidenced that the fraction of FAA, which represented <2% of dry matter of the feeds, is quickly released in water, acting mainly as attractants. The in vitro digestibility assay evidenced significant differences in the bioaccessibility of the nitrogen fraction in some of the feeds by the digestive proteases of the sea bream (Sparus aurata).