1-O-Alkylglycerol Ethers from the Marine Sponge Guitarra abbotti and Their Cytotoxic Activity

The cytotoxicity-bioassay-guided fractionation of the ethanol extract from the marine sponge Guitarra abbotti, whose 1-O-alkyl-sn-glycerol ethers (AGEs) have not been investigated so far, led to the isolation of a complex lipid fraction containing, along with previously known compounds, six new lipids of the AGE type. The composition of the AGE fraction as well as the structures of 6 new and 22 previously known compounds were established using ¹H and ¹³C NMR, GC/MS, and chemical conversion methods. The new AGEs were identified as: 1-O-(Z-docos-15-enyl)-sn-glycerol (1), 1-O-(Z-docos-17-enyl)-sn-glycerol (2), 1-O-(Z-tricos-15-enyl)-sn-glycerol (3), 1-O-(Z-tricos-16-enyl)-sn-glycerol (4), 1-O-(Z-tricos-17-enyl)-sn-glycerol (5), and 1-O-(Z-tetracos-15-enyl)-sn-glycerol (6). The isolated AGEs show weak cytotoxic activity in THP-1, HL-60, HeLa, DLD-1, SNU C4, SK-MEL-28, and MDA-MB-231 human cancer cells. A further cytotoxicity analysis in JB6 P⁺ Cl41 cells bearing mutated MAP kinase genes revealed that ERK2 and JNK1 play a cytoprotective role in the cellular response to the AGE-induced cytotoxic effects.     

Comparative pharmacokinetics of enrofloxacin and ciprofloxacin in lactating dairy cows and beef steers following intravenous administration of enrofloxacin

The comparative pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin were investigated in lactating cows and beef steers. The plasma elimination half-life of either enrofloxacin or ciprofloxacin was shorter in cows than in steers. The overall production of ciprofloxacin was slightly higher in steers than in cows (metabolite ratio: 64% and 59%, respectively). There was no significant difference in plasma protein binding of enrofloxacin between cows (percent bound: 59.4%) and steers (percent bound: 60.8%). Ciprofloxacin was more extensively bound to plasma proteins in steers (percent bound: 49.6%) than in cows (percent bound: 33.8%). The steady state volume of distribution of enrofloxacin is comparable in cows (1.55 L/kg) and steers (1.59 L/kg). Within either bovine class, plasma elimination half-life of enrofloxacin and ciprofloxacin are comparable, while plasma protein binding was higher for enrofloxacin than for ciprofloxacin. Ciprofloxacin was more concentrated in milk than enrofloxacin.     

Population Structure of Phytophthora cinnamomi in South Africa

ABSTRACT Phytophthora cinnamomi isolates collected from 1977 to 1986 and 1991 to 1993 in two regions in South Africa were analyzed using isozymes. A total of 135 isolates was analyzed for 14 enzymes representing 20 putative loci, of which four were polymorphic. This led to the identification of nine different multilocus isozyme genotypes. Both mating types of P. cinnamomi occurred commonly in the Cape region, whereas, predominantly, the A2 mating type occurred in the Mpumalanga region of South Africa. A2 mating type isolates could be resolved into seven multilocus isozyme genotypes, compared with only two multilocus isozyme genotypes for the A1 mating type isolates. Low levels of gene (0.115) and genotypic (2.4%) diversity and a low number of alleles per locus (1.43) were observed for the South African P. cinnamomi population. The genetic distance between the Cape and Mpumalanga P. cinnamomi populations was relatively low (D(m) = 0.165), and no specific pattern in regional distribution of multilocus isozyme genotypes could be observed. The genetic distance between the "old" (isolated between 1977 and 1986) and "new" (isolated between 1991 and 1993) P. cinnamomi populations from the Cape was low (D(m) = 0.164), indicating a stable population over time. Three of the nine multilocus isozyme genotypes were specific to the "old" population, and only one multilocus isozyme genotype was specific to the "new" population. Significant differences in allele frequencies, a high genetic distance (D(m) = 0.581) between the Cape A1 and A2 mating type isolates, significant deviations from Hardy-Weinberg equilibrium, a low overall level of heterozygosity, and a high fixation index (0.71) all indicate that sexual reproduction occurs rarely, if at all, in the South African P. cinnamomi population.     

Horses' behavior and heart rate in a preference test for shorter and longer riding bouts

A Y-maze preference test was used to investigate whether horses prefer a shorter riding treatment over a longer riding treatment. In a pilot study (n = 4 horses), the maze was positioned with the 2 arms each pointing toward one of the short sides of the indoor arena, and in the main study (n = 14), the maze was rotated by 90°. Horses were 11 ± 4.4 years old and ridden regularly for at least 5 times/wk. They were conditioned to associate 1 exit of the maze with 1 lap of riding (R1), covering a distance of approximately 40 m at the walk or trot according to a predetermined schedule, and the other exit with 2 such laps (R2). Immediately afterward, riders dismounted, horses were led into the maze, and horses were let loose to make their choice in the maze. After exiting the maze, the rider mounted again and rode according to the chosen treatment. This procedure was repeated on the same day until statistical significance (P < 0.05) of preference was reached or up to a maximum of 35 trials. In addition, behavioral observations and heart rates were recorded. In the pilot study, all horses, regardless of the associated treatment, chose the left arm, which, unfortunately, pointed toward the arena's exit door. If horses were not caught immediately after exiting the maze (n = 5 occasions), they walked or trotted straight to the door. In the main study, 4 horses significantly preferred R1, 2 horses significantly preferred R2, and 8 horses had no significant preference. Heart rates were significantly (P < 0.05) higher during R2 (87.4 ± 2.6 bpm) than during R1 (79.5 ± 2.4 bpm). Except for tail swishing, no significant differences were found for the frequency of occurrence of behavior patterns between R1 and R2. Over the course of repeated trials, some horses became increasingly reluctant to enter and walk through the maze, and most showed increasing resistance to being remounted (e.g., sidestepping). Overall, the experimental setup did not seem to be appropriate to answer the research question. It is likely that the repeated mounting and dismounting caused discomfort or confused many of the horses to an extent that they did not actively select a treatment but rather searched for ways to evade further mounting (and riding). In conclusion, horses did not show a clear preference for either shorter or longer riding bouts, but their behavioral reactions indicate that they perceived mounting as uncomfortable and that their motivation to rejoin their herd-mates and/or to obtain feed in the barn was greater than their motivation to being ridden at all. Nevertheless, pronounced individual differences also seem to exist, with some horses showing little aversion to, and perhaps enjoyment of, being ridden, whereas others clearly preferring not to be ridden.     

Evaluation of Methods to Determine Sperm Density for the European eel,Anguilla anguilla

European eel, Anguilla anguilla, is a target species for future captive breeding, yet best methodology to estimate sperm density for application in in vitro fertilization is not established. Thus, our objectives were to evaluate methods to estimate European eel sperm density including spermatocrit, computer‐assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore, accuracy and precision of spermatocrit, applying a range of G‐forces, were tested and the best G‐force used in method comparisons. We found no effect of dilution on haemocytometer sperm density estimates, whereas motility associated positively with haemocytometer counts, but not with spermatocrit. Results from all techniques, spermatocrit, CASA and FCM, showed significant positive correlations with haemocytometer counts. The best correlation between spermatocrit and haemocytometer counts was obtained at 6000 ×  g (r = 0.68). Of two CASA variants, one or three photographic fields (CASA‐1 and CASA‐2), CASA‐2 showed a very high accuracy to haemocytometer counts (r  =  0.93), but low precision (CV: CASA‐2 = 28.4%). FCM was tested with and without microfluorospheres (FCM‐1 and FCM‐2), and relationships to haemocytometer counts were highly accurate (FCM‐1: r  =  0.94; FCM‐2: r  =  0.88) and precise (CV: FCM‐1 = 2.5; FCM‐2 = 2.7%). Overall, CASA‐2 and FCM‐1 feature reliable methods for quantification of European eel sperm, but FCM‐1 has a clear advantage featuring highest precision and accuracy. Together, these results provide a useful basis for gamete management in fertilization protocols.     

A comparison of aggressiveness and deoxynivalenol production between Canadian <Emphasis Type="Italic">Fusarium graminearum</Emphasis> isolates with 3-acetyl and 15-acetyldeoxynivalenol chemotypes in field-grown spring wheat

Twenty four isolates of Fusarium graminearum, half of which were 3-acetyldeoxynivalenol (3-ADON) and half 15-acetyldeoxynivalenol (15-ADON) chemotypes, were tested for their ability to produce deoxynivalenol and to cause Fusarium head blight (FHB) in spring wheat cultivars. The objectives of this study were to determine (1) whether 3-ADON isolates differ in aggressiveness, as measured by the FHB index, and DON production from 15-ADON isolates under field conditions, and (2) whether the performance of resistant host cultivars was stable across isolates. Field tests of all isolates were conducted with three replicates at each of two locations in Canada and Germany in 2008 with three host genotypes differing in FHB resistance level. The resistant host genotype showed resistance regardless of the chemotype or location. The differences between mean FHB indices of 3-ADON and 15-ADON isolates were not significant for any wheat genotype. In contrast, average DON production by the 3-ADON isolates (10.44 mg kg⁻¹) was significantly (P < 0.05) higher than for the 15-ADON isolates (6.95 mg kg⁻¹) at three of the four locations where moderately resistant lines were tested, and at both locations where susceptible lines were evaluated. These results indicate that 3-ADON isolates could pose a greater risk to food safety. However, as the mean aggressiveness and DON production of 3-ADON and 15-ADON chemotypes was similar on highly resistant lines, breeding and use of highly resistant lines is still the most effective measure of reducing the risks associated with DON in wheat.     

Dissolved and colloidal phosphorus fluxes in forest ecosystems—an almost blind spot in ecosystem research

Understanding and quantification of phosphorus (P) fluxes are key requirements for predictions of future forest ecosystems changes as well as for transferring lessons learned from natural ecosystems to croplands and plantations. This review summarizes and evaluates the recent knowledge on mechanisms, magnitude, and relevance by which dissolved and colloidal inorganic and organic P forms can be translocated within or exported from forest ecosystems. Attention is paid to hydrological pathways of P losses at the soil profile and landscape scales, and the subsequent influence of P on aquatic ecosystems. New (unpublished) data from the German Priority Program 1685 “Ecosystem Nutrition: Forest Strategies for limited Phosphorus Resources” were added to provide up‐to‐date flux‐based information. Nitrogen (N) additions increase the release of water‐transportable P forms. Most P found in percolates and pore waters belongs to the so‐called dissolved organic P (DOP) fractions, rich in orthophosphate‐monoesters and also containing some orthophosphate‐diesters. Total solution P concentrations range from ca. 1 to 400 µg P L^?1, with large variations among forest stands. Recent sophisticated analyses revealed that large portions of the DOP in forest stream water can comprise natural nanoparticles and fine colloids which under extreme conditions may account for 40–100% of the P losses. Their translocation within preferential flow passes may be rapid, mediated by storm events. The potential total P loss through leaching into subsoils and with streams was found to be less than 50 mg P m^?2 a^?1, suggesting effects on ecosystems at centennial to millennium scale. All current data are based on selected snapshots only. Quantitative measurements of P fluxes in temperate forest systems are nearly absent in the literature, probably due to main research focus on the C and N cycles. Therefore, we lack complete ecosystem‐based assessments of dissolved and colloidal P fluxes within and from temperate forest systems.     

Seasonal effects of liming,irrigation, and acid precipitation on microbial biomass N in a spruce (Picea abies L.) forest soil

Summary Seasonal effects of liming, irrigation, and acid precipitation on microbial biomass N and some physicochemical properties of different topsoil horizons in a spruce forest (Picea abies L.) were measured throughout one growing season. The highest biomass N was recorded in autumn and spring in the upper soil horizons, while the lowest values were obtained in summer and in deeper horizons. The clearest differences between the different soil treatments were apparent in autumn and in the upper horizons. Liming increased the microbial biomass N from 1.7% of the total N content to 6.8% (Olf₁ layer) and from 1% to 2% of the total N content in the Of₂ layer. The main inorganic-N fraction in the deeper horizons was NO _inf3 ^sup- . An increase in cation exchange capacity was observed down to the Oh layer, while soil pH was only slightly higher in the Olf₁ and Of₂ layers after liming. The effects of irrigation were less marked. The microbial biomass N increased from 1.7% of total N to 4.8% in the Olf₁ layer and from 1% to 2% of total N in the Of₂ layer. In the Olf₁ layer an increase in C mineralization was observed. Acid precipitation decreased the microbial biomass N in the upper horizons from 4.8% of total N to 1.8% in the Olf₁ layer and from 2% to 0.5% in the Of₂ layer. No significant changes in soil pH were observed, but the decrease in cation exchange capacity may result in a decrease in the proton buffering capacity in the near future.