Isolation and cryopreservation of functionally competent equine leucocytes

Sufficient numbers of functionally competent polymorphonuclear neutrophil granulocytes (PMN) seem to be of major importance during the course of equine endometritis. In this study, we wanted to establish a method for cryopreservation of functionally competent neutrophils for an intended local endometritis therapy in mares. The separation of leucocytes by hypotonic lysis of whole blood from clinically healthy mares was superior to the separation by dextrose sedimentation. After suspension of the cells in the cryoprotective solution [equine plasma with 5% (v/v) dimethyl sulphoxide (DMSO)], the leucocytes were frozen in liquid nitrogen. A temperature gradient with low cooling velocity (1 degree C/min between 4 and -70 degrees C) resulted in highest numbers of viable cells after thawing. Thawed PMN had a high phagocytic capacity for opsonized streptococci. Their ability to generate reactive oxygen species (ROS) after stimulation with a phorbol ester was even higher than that of freshly isolated PMN and was preserved up to 6 h after thawing. The results of this study indicate that cryopreservation of PMN may provide viable and functionally competent neutrophils for therapeutic use in mares susceptible to endometritis.     

Efficacy of milbemycin oxime in the treatment of canine generalized demodicosis: a retrospective study of 99 dogs (1995-2000)

Ninety-nine dogs diagnosed with generalized demodicosis were treated with milbemycin oxime. Cases diagnosed before two years of age were classified as juvenile-onset (53%) and those diagnosed after two years of age as adult-onset (47%). Dogs were considered cleared of mites when none could be demonstrated in scrapings and cured if no relapse was seen for 12 months. Eighty-five per cent (84/99) were cured with milbemycin oxime used for 1-6 months (mean 2.3 months) at a dosage of 0.5-1.6 mg kg-1 body weight (mean 0.75 mg kg-1). No significant difference in dosage or treatment time was seen between juvenile and adult cases. Chance of cure was significantly better in young animals (cured cases mean age 2.97 years) than in older animals (not cured cases mean age 8.02 years). Start of treatment early in the course of disease gave a significantly better chance of cure. Cases with severe pododemodicosis had less chance of a cure (9/11 not cured).     

"Emergency splitting" of the teat canal--a therapeutic procedure in case of concurrent occurrence of acute thelitis and severe mastitis

In case of concurrent acute thelitis and severe mastitis, an immediate surgery of the stenosis ("emergency splitting") might be justified in order to improve milkability as a prerequisite for the removal of pathological milk secretion and therapy, successful mastitis therapy. The aim of this study was to compare the milkability and the clinical cure rate of mastitis between Group A (n = 19, cows with stenosis, thelitis and mastitis) and Group B (n = 19, cows with stenosis, with or without clinical mastitis, but without acute thelitis) following surgery of the papillary duct using a cutting instrument (Danish cannula model with a double cutting edge). In most cases, milkability was restored at the end of the treatment in both groups (Group A: 89.5%; Group B: 100%). In this time period nine of 19 quarters with mastitis in Group A were clinically cured (47%). Six month after the surgery, the animal owners considered the milkability as very good in 70% and 72% of the cases in Group A and Group B, respectively. However, "emergency splitting" led to a more intensive and longer postsurgical treatment. The average duration of total treatment (day of surgery plus 6 days of obligate postsurgical treatment plus additional treatment days) was 7.4 and 6.7 days in Group A and Group B, respectively. Furthermore, the higher expenditure was due to the more costly mastitis therapy. During the six month following surgery, only one cow of Group A had to be culled due to persistent mastitis and two cows because of insufficient milkability.     

Influence of different forest conversion strategies on ground vegetation and tree regeneration in pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) stands: a case study in NE Germany

The conversion of anthropogenic into more natural, self-regenerating forests is one of the major objectives of forestry throughout Europe. In this study, we present investigations on permanent plots with different silvicultural treatment in NE German pine stands. Management of old-growth pine stands on acidic and nutrient-poor sandy sites differs in fencing, thinning, and planting of certain tree species. The investigations were carried out on the community, population, and individual level of the pine forest ecosystems. Thus, vegetation changes, size and height of tree populations, and height increment of tree individuals were observed over a time span of 6 years. Special attention was paid to short-lived tree species such as, e.g., Frangula alnus and Sorbus aucuparia, as well as to Fagus sylvatica as one of the most typical forest tree species of Central Europe. Vegetation changes are interpreted as a consequence of natural regeneration of formerly degraded forest sites, involving an increase in nutrient availability. High browsing pressure can be considered as a key factor for the inhibition of tree seedlings and growth of saplings. Some Sorbus aucuparia individuals, however, succeeded in growing out of the browsing height also in unfenced stands. Few found specimens of Fagus sylvatica proved that this species is able to establish spontaneously on these relatively dry, acidic sites under continental climate influence. Such natural regeneration processes, also including spontaneous rejuvenation of trees, can be integrated into silviculture as passive forest conversion management. An active management like thinning of stands, planting of trees, and fencing can accelerate forest conversion with regard to height growth and species number of trees.     

Suppression of C-hordein synthesis in barley by antisense constructs results in a more balanced amino acid composition

Barley has for feeding purposes a shortage of essential amino acids, especially lysine, threonine, and methionine, and an excess of proline and glutamine. In the present study, we have introduced into barley an antisense construct against C-hordeins, the storage protein with the lowest nutritional quality. SDS-PAGE and reverse phase HPLC revealed a relative reduction in the amounts of C-hordeins and relative increases in the content of the other storage proteins. The five different lines analyzed had lower amounts of proline, glutamic acid/glutamine, and phenylalanine (up to 12%, 6%, and 9% reductions), while the lysine, threonine, and methionine content was increased with up to 16%, 13% and 11%. It is concluded that antisense mediated suppression of C-hordein synthesis may be a promising approach for improving the nutritional value of barley as a feed crop while at the same time reducing the environmental nitrogen load.     

A method for identification and mapping potentially acid sulfate soils in Jutland, Denmark

A method for mapping potentially acid sulfate soils in Jutland, Denmark is described. First the wetlands were outlined, using already existing topographic maps. Then 7900 profiles, evenly distributed over the wetland areas, were examined to point out areas with potentially acid sulfate soils. The method used to identify potentially acid sulfate material was, for carbonate-free samples, a pH-measuring at the sampling time and after the samples were inoculated with soil water extract from an acid sulfate soil and 2, 8 or 16 weeks of incubation. For carbonate-containing samples the pyrite content was compared to the acid-neutralizing capacity. A map was constructed showing the ratio of profiles containing potentially acid sulfate soil samples to total number of profiles examined within natural geographical wetland regions.The investigation showed that 35% of the wetlands in Jutland have a high frequency of potentially acid sulfate soils.     

New mosquito larvicidal tetranortriterpenoids from Turraea wakefieldii and Turraea floribunda

The crude methanol extracts of the root barks of Turraea wakefieldii and Turraea floribunda were found to show mosquito larvicidal activity against third-instar larvae of Anopheles gambiae sensu stricto. Four new limonoids comprising a vilasininoid 1 and three havanensinoids 2-4 were isolated from the chloroform fractions of the methanol extracts of T. wakefieldii and T. floribunda, respectively. The structures of the compounds were elucidated by NMR spectroscopy. Compounds 1, 2, and 4 had LD50 values of 7.1, 4.0, and 3.6 ppm, respectively, and were more potent than azadirachtin, which had an LD50 value of 57.1 ppm when tested against larvae of A. gambiae.     

Quality of different mulch materials and their decomposition and N release under low moisture regimes

During the dry season in the tropics, agriculture which is solely dependent upon rainfall as its source of water is frequently affected by soil moisture stress, resulting in crop failures. Farmers therefore depend mainly on other sources of limited water supply during this period, such as ground water. Soil moisture conservation measures, especially surface mulching with loppings and, occasionally, leaf litter and crop residues, are practised. Our objective was to study the decomposition and nitrogen (N) release from these plant materials under continuously wet, low moisture regimes, i.e. comparable to those which prevail in the mulches used in the agriculture. A greenhouse experiment was conducted with fresh, chopped leaves of six leguminous trees, wild sunflower and rice, which were spread as a mulch on a layer of soil. They were maintained at eight moisture levels (a total of between zero and 43l water m^–2 applied over 8 weeks) by spraying water. Different optimal moisture requirements for the rapid decomposition of these species were observed. These were presumably determined by different physical and chemical properties of the leaves. The amount of water received to the mulches and their soluble polyphenolic and carbon (C) concentrations played an important role in determining the decomposition and the mode of N release under non-limiting conditions of leaf N. Specifically, the C concentration governed N release, while the effect of polyphenolics was important when their concentration was low, as a result of leaching under relatively high moisture regimes. Leaves with a high polyphenolic and C content, which were subjected to high leaching losses of these fractions, underwent a change in their N dynamics from net immobilization to mineralization. This study indicates that leaves with a fast rate of decomposition should be mixed with other species, leaves which decompose more slowly in order to increase the conservation of soil moisture and also improve the synchronization between N release from the mulch and its demand by crops. Received: 6 January 1997     

Element (Ag, Cd, Cu, Pb, Sb, Tl and Zn), element ratio and lead isotope profiles in a sediment affected by a mining operation episode during the late 19th century

Mining operations at Mårsätter in 1877–81 resulted in increased metal loading to a small lake, notably as sulphidic tailings. The event is taken as an opportunity to study the present environmental impact of a historical single major metal release. Lake water and four sediment cores were sampled and analysed for principal and trace elements in solid and aqueous phases as well as general hydrochemical conditions. Chronologies were determined from ²⁰⁶Pb/²⁰⁷Pb ratios and historical records. Ordinary sedimentation after the event has lead to that the tailings are found as a distinct layer at a depth of 18–22 cm in the sediment. The layer is characterized by elevated metal concentrations in the solid and pore water phases, respectively, circum neutral pH and sulphate concentrations below detection. Geochemical modelling indicated a preference for carbonate equilibrium in the waste while sulphides prevailed above it. It is concluded that the growth of the ordinary sediment on top of the waste has lead to a physicochemical barrier that seals of the waste from the overlying sediment. Chemical or physical rupture of the barrier will release the metals to downstream regions. According to the chronologies at least three sources have contributed to the present elevated levels of metals, in additions to the release of tailings. Copper from a historical blast furnace prior to the event at Mårsätter, transport from mineralized parts of the watershed and release of contaminated water from present mining operations maintain elevated levels of notably zinc, silver, cadmium and lead. At present less than 10% of the lead content at the sediment/water interface comes from atmospheric deposition. Increased levels of antimony and thallium were not observed prior to ca 1950.