Chemical composition and antimicrobial and antioxidant activities of the essential oil and methanol extract of Hippomarathrum microcarpum (Bieb.) from Turkey

Hippomarathrum microcarpum grows wild in eastern Anatolia, Turkey, and is a plant utilized as food by people. In this study, the in vitro antimicrobial and antioxidant activities of the essential oil and methanol extract from H. microcarpum and its essential oil composition were investigated. The essential oil, which has bornyl acetate, caryophyllene oxide, and beta-caryophyllene as its main components, exhibited activity against eight bacteria, nine fungi, and a yeast, Candida albicans, with minimum inhibitory concentration values ranging from 62.50 to 125 muL/mL; the methanol extract showed weak activity. The antioxidant activity of these extracts was assessed by the beta-carotene bleaching test and the 1,1'-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging test. The inhibition of linoleic acid oxidation was very weak for both extracts tested. The inhibition percentages were found to be 22.9 and 33.5% for methanol and essential oil, respectively, at the concentration of 2 g/L. The oil scavenged DPPH at higher concentrations (IC50 = 10.69 +/- 0.05 mg/mL), but the methanol extract exhibited no activity. The total phenolic content of the methanol extract was found to be 4.7 +/- 0.1%.     

Use of thermal imaging to determine leaf conductance along a canopy gradient in European beech (Fagus sylvatica)

Using an infrared camera, we measured the leaf temperature across different canopy positions of a 23-m-tall deciduous forest tree (Fagus sylvatica L.) including typical sun and shade leaves as well as intermediate leaf forms, which differed significantly in specific leaf area (SLA). We calculated a temperature index (I(G)) and a crop water stress index (CWSI) using the surface temperatures of wet and dry reference leaves. Additional indices were computed using air temperature plus 5 °C (I(G)?+?5, CWSI?+?5) as dry references. The minimum temperature of the wet leaf and the maximum temperature of the dry leaf proved to be most suitable as reference values. We correlated the temperature indices with leaf area-related conductance to water vapor (g(L)) using porometry at the leaf level and using xylem sap flow at the branch level. At the leaf and at the branch level, I(G) and CWSI were equally well suited as proxies of g(L), whereas the relationships of I(G)?+?5 and CWSI?+?5 with g(L) were only weak or even insignificant. At the leaf level, the correlations of I(G) and CWSI with g(L) were significant in all parts of the crown. The slopes of g(L) vs. I(G) and CWSI did not differ significantly among the crown parts; this indicates that they were not influenced by SLA or irradiance. At the branch level, close correlations (r?>?0.8) were found between temperature indices and g(L) across the crown. These results demonstrate that satisfactory relationships between temperature indices and g(L) can be established in tall trees even in those canopy parts that are exposed to relatively low levels of irradiance and exhibit relatively low values of g(L).     

Emended descriptions of indole negative and indole positive isolates of Brachyspira (Serpulina) hyodysenteriae

Two type/reference strains of Brachyspira (B.) hyodysenteriae, 14 Belgian and German indole negative, and 14 Belgian, German and Swedish indole positive field isolates of strongly β-haemolytic intestinal spirochaetes were compared by pulsed-field gel electrophoresis (PFGE) patterns, biochemical reaction patterns, 16S rDNA sequences and MIC determinations of six antibacterial substances. Three tests for indole production, including a spot indole test, were compared with congruent results. All field isolates were classified as B. hyodysenteriae due to a high genetic and phenotypic similarity with the type strains. The Belgian and German indole negative isolates had identical and unique PFGE patterns for the tested restriction enzymes MluI and SalI, as well as identical 16S rDNA sequences, and they could not be differentiated by any of the methods used. Seven unique PFGE patterns were achieved from the 14 indole positive field isolates. The patterns were identical and unique for epidemiologically related isolates. Type/reference strains and isolates without known relation to other tested isolates showed unique banding patterns. The MICs of tylosin, tiamulin, erythromycin, clindamycin, carbadox and virginiamycin were determined in broth for all isolates. In contrast to Belgian and German isolates, the majority of the Swedish field isolates were susceptible to tylosin, erythromycin and clindamycin. Probable pathways of infection for some of the Swedish isolates were determined. The PFGE patterns of epidemic clones of B. hyodysenteriae remained stable for a period of up to 8 years. In vivo development of resistance to macrolide and lincosamide antibiotics due to use of tylosin was clearly indicated for two epidemic clones.     

New screening test to predict the potential impact of ivermectin-contaminated cattle dung on dung beetles

According to European Union recommendations, a test method has been developed to evaluate the effects of veterinary pharmaceuticals on dung feeding insects. This test method was evaluated with the dung beetle Aphodius constans by using fecal residues of ivermectin after a pour-on administration. Dung of different age (and thus containing different concentrations of ivermectin) as well as mixtures of highly-contaminated spiked dung with untreated control dung were studied in five test runs in two laboratories. The concentration of ivermectin (active substance; a.s.) in the dung samples was verified analytically. The main test endpoint was the survival of first instar larvae. The LC50 using dung directly obtained from treated cattle ranged from 470 to 692 microg a.s. kg(-1) dung (dry weight; d.w.) and 67 to 97 microg a.s. kg(-1) dung (fresh weight; f.w.). Using mixtures, the outcome of two tests was almost identical: 770 to 781 microg a.s. kg(-1) dung (d.w.); 109 to 132 microg a.s. kg(-1) dung (f.w.). In comparison to the LC50 values obtained when ivermectin was spiked in control dung at several concentrations (LC50 880-985 microg a.s. kg(-1) dung (d.w.)), the LC50 values were again very similar. Three conclusions can be drawn from these results. The proposed test method seems to be robust and allows for the initiation of an international validation process (including ringtesting). Because of only small differences found in tests in which the test substance was spiked into control dung and those in which dung from treated cattle was applied, the use of a standard test method is proposed. The effects of ivermectin on ecologically relevant dung beetles obtained in a standardised test method reflect the results from field studies and are in the range of environmentally relevant concentrations.     

The effect of treatment with eprinomectin on lungworms at early patency on the development of immunity in young cattle

An experiment was carried out to study the effect of topical application of eprinomectin at early patency on the build up of infection and development of protection against Dictyocaulus viviparus in young cattle. Three groups of six calves were used and parasitological and blood variables were monitored at weekly intervals throughout the trial. At the start of the experiment calves in groups A and B were experimentally inoculated with 100 D. viviparus infective third-stage larvae (L3) for five consecutive days, whereas calves in group C served as uninfected controls. The calves in group A were each treated with eprinomectin (0.5mg/kg bodyweight) in a pour-on formulation at early patency at day 24 post the first inoculation, whereas the calves in groups B and C were left untreated. Seven weeks following anthelmintic treatment all groups were challenged with 1500 L3. Another 4 weeks later the animals were sacrificed and established worms in the lungs were counted. Moderate transient signs of lungworm disease occurred both in groups A and B. However, group B calves were found to be about 8 times more resistant than those in group A, whereas the naive infection controls in Group C was found to be about 35 times more susceptible to infection. Also the ELISA values showed that the course of infection was different between experimental groups. The eosinophil counts prior to and at the time of slaughter indicate that immunity was involved in the protection and the response was correlated with previous exposure and worm load. Weight gains differed significantly, but only between groups A and C and between groups B and C that on an average were approximately 13kg heavier at the termination of the experiment. It was concluded that eprinomectin was effective against established adult lungworms. However, the untreated calves (group B) developed a more marked resistance to lungworms compared to those that were subjected to anthelmintic treatment at early patency (group A). On the other hand, the cumulative number of excreted larvae was on an average 43 times higher in group B as compared to group A. Consequently, infected calves that remain out on pasture should be treated. This will restrain transmission of the parasite despite the fact that immunity is deteriorated.     

Genetic diversity in seven populations of Nicaraguan teosinte (<Emphasis Type="Italic">Zea nicaraguensis</Emphasis> Iltis et Benz) as estimated by microsatellite variation

Teosintes are the closest relatives to modern maize, Zea mays L. ssp. mays. They are wild grasses with a native distribution area from Mexico to Nicaragua and represent an important genetic resource. However, the genetic diversity of Nicaraguan teosinte (Zea nicaraguensis Iltis et Benz) has not yet been determined. This teosinte species has decreased in the last 25 years and now must be regarded as an endangered species. An analysis of the genetic diversity of Zea nicaraguensis was carried out in a total of 240 individuals from seven populations. Eleven Simple Sequences Repeat (SSR) primer pairs were used. A total of 42 alleles were found, the range of alleles per locus was 2–5 (mean 3.8) and the numbers of genotypes varied between primers. The primer Bnlg 1538 showed the highest value, with 45 genotypes through all populations. The genetic diversity observed (Ho) between all populations varied from 0.51 to 0.63, with an average of 0.563. One of the populations had as many as 40 alleles. Analysis of molecular variance (AMOVA) revealed that most of the variation was within population, at a significantly high level (P < 0.001). Rare alleles were detected in all populations, but unique alleles were only found in four populations. These results are highly relevant when developing conservation strategies and show that preserving populations in their natural habits is highly important.