Experimental studies on cold survival of enchytraeid cocoons

The effect of different culture methods, cold acclimation and desiccation on the supercooling point (SCP), the melting point (MP) of fluids and the quantity of water freezing (osmotically active water) was investigated in individual cocoons of Enchytraeus crypticus and an undescribed Enchytraeus species using differential scanning calorimetry (DSC) techniques. Both species can be easily cultured in the laboratory in agar where the development and hatching of the worms can be observed. Culture methods (agar with nettle leaves or oats as food and wet filter paper without food supply) had a significant effect on fresh weight and SCP of E. crypticus cocoons. The water content (as a proportion of fresh weight) was slightly lower in the cocoons from the wet filter paper cultures. Acclimation at —3 °C did not affect the supercooling ability of E. crypticus cocoons, whereas the SCP of E. sp.1 cocoons was lowered from a mean of —8.7 to —12.4 °C. The supercooling ability of cocoons of E. crypticus was only slightly increased by desiccation and cold acclimation, but the quantity of frozen water was significantly reduced with acclimation and desiccation (2 and 3 min) at room temperature. The MP values of the cocoon fluids reflected these changes in both species. No cocoons of E. crypticus and E. sp.1 survived desiccation and freezing in our experiments and no worms of Enchytraeus albidus, E. crypticus and Henlea perpusilla survived freezing to —10 or —20 °C.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Einfluß von Schwefeldioxid auf Fichten (Picea abies [L.] Karst.) in verschiedenen Bodensubstraten

Zusammenfassung Drei Jahre alte Fichten wurden nach einer sechsmonatigen Anzuchtsphase in verschiedenen Bodensubstraten sieben Wochen lang mit 140 μg SO₂/m³ begast. Dabei kam ein Begasungsprinzip zur Anwendung, das auch bei Dauerbelastung konstante Gaskonzentrationen, unabh?ngig von sich ?ndernden Depositionsraten gew?hrleistet und Gaswechselmessungen in der geschlossenen Anzuchts- und Begasungskammer erlaubt. Es zeigte sich, da? sowohl n?hrstoffarme B?den als auch eine SO₂-Belastung im Kronenbereich zu Beeintr?chtigungen des Mineralstoffhaushaltes der Pflanzen führen. Insbesondere in den Wurzeln kam es nach einer siebenw?chigen Begasungsdauer zu einer starken Verminderung der Calcium- und Magnesiumgehalte. Auch die Photosyntheseleistung der Pflanzen und der Chlorophyllgehalt der Nadeln wurde durch die kombinierte Belastung mit sauren, n?hrstoffarmen Substraten und Schwefeldioxid st?rker beeintr?chtigt als durch den jeweiligen Einflu? der einzelnen Faktoren.

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Effects of sulfur dioxide on spruces (Picea abies [L.] Karst.) growing in different soil substrata Exploratory investigations on photosynthesis and mineral nutrient supplies following long-term application of low SO₂ concentrations

Summary Three-year-old spruce plants were grown in different soils for six months and then treated with 140 μg SO₂/m³ for seven weeks. The fumigation system used provides constant gas concentrations during a long-term fumigation period. Information on the combined growth and fumigation chamber is presented. It can be shown that not only nutrient deficiency in the soil but also dry deposition of SO₂ affects the mineral supply of plants. In particular the contents of calcium and magnesium in the roots were drastically reduced after the 7-week exposure period. The combination of acid soil and sulfur dioxide had a greater effect on both net photosynthetic rates of the plants, and chlorophyll contents of the needles as compared with the effect of only one of these stresses.

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Herrn Prof. Dr.W. Liese zum 60. Geburtstag gewidmet.     

Content of polyphenolic compounds in the Nigerian stimulants Cola nitida ssp. alba, Cola nitida ssp. rubra A. Chev, and Cola acuminata Schott & Endl and their antioxidant capacity

Varieties of kola nuts (Cola nitida alba, Cola nitida rubra A. Chev, and Cola acuminata Schott & Endl), a group of popular Nigerian and West African stimulants, were analyzed for their content of secondary plant metabolites. The three varieties of the kola nuts contained appreciable levels of (+)-catechin (27-37 g/kg), caffeine (18-24 g/kg), (-)-epicatechin (20-21 g/kg), procyanidin B 1 [epicatechin-(4beta-->8)-catechin] (15-19 g/kg), and procyanidin B2 [epicatechin-(4beta-->8)-epicatechin] (7-10 g/kg). Antioxidant capacity of the extracts and purified metabolites was assessed by two HPLC-based and two colorimetric in vitro assays. Extracts of all varieties exhibited antioxidant capacity with IC 50 values in the range 1.70-2.83 and 2.74-4.08 mg/mL in the hypoxanthine/xanthine oxidase and 2-deoxyguanosine HPLC-based assays, respectively. Utilization of HPLC-based assays designed to reflect in situ generation of free radicals (e.g., HO(*)), as opposed to general assays (DPPH, FRAP) in common use which do not, indicate that, of the major secondary plant metabolites present in kola nut extracts, caffeine is potentially the more effective cancer chemopreventive metabolite in terms of its antioxidant capacity.     

A method of preparing mesocosms for assessing complex biotic processes in soils

Equipment and handling methods for the preparation of soil mesocosms were developed. The mesocosms were used to investigate the interrelationships between mesofauna and microflora in a coniferous forest soil. Soil monoliths were taken from the ground, defaunated by deep-freezing, wrapped in nets to control reimmigration of different faunal size-classes, and replanted in the field for 8 months. in a practical test the technique described here proved to be an inexpensive field method for producing a replicated series of mesocosm in a short time. Deep-freezing is appropriate for defaunating soil monoliths. The fine nets effectively exluded meso-and macrofauna. No significant differences were found in the abundance of Enchytraeids and Collembola between recolonized mesocosms and the undisturbed control at the end of the study period. In contrast, oribatid mite abundace was still greatly reduced in the recolonized esocosms. Dominance structure and species composition of the more dominant oribatid species in the different treatments were apparently similar. To compensate for the low colonization ability of oribatids, a reintroduction of selected animal size-classes to defaunated monoliths is recommended.     

Chloroplast DNA polymorphism of an exotic P. mugo Turra population introduced to seaside spit of Kursiu Nerija in Lithuania

The aim of this study was to elucidate the introduction history of P. mugo in the unique landscape of the Lithuanian seaside spit of Kursiu Nerija by assessing its genetic structure and the genetic diversity. The individuals were sampled in 12 populations within an area of 3 km × 50 km along the Lithuanian part of Kursiu Nerija. P. mugo was introduced over 200 years ago to prevent sand erosion by establishing a forest cover. Chloroplast DNA polymorphism of 220 individuals of P. mugo together with 18 P. sylvestris and 11 putative P. sylvestris × P. mugo hybrids was assessed by the aid of five microsatellite markers. The standard intra-population diversity indexes were calculated. The intra-specific variation between distinct morphotypes as well as the population differentiation within the most spread P. mugo ssp. rotundata morphotype was assessed based on the haplotype frequencies by hierarchical AMOVA, G_ST/R_ST test, UPGMA clustering and PCA methods. The genetic diversity of P. mugo in Kursiu Nerija was high (H_e = 0.95; 83 different haplotypes). All except one of the P. mugo populations sampled contained a notable share of private haplotypes. AMOVA revealed high intra-specific diversity but low differentiation between the P. mugo populations. Most of the haplotypic variance was within populations. The UPGMA clustering produced groups more corresponding to the sub-species morphotypes than the geography of the populations. There was no geographical pattern of reduction in genetic diversity towards the younger plantations. A strong candidate for a species-specific DNA marker was found. After several events of introduction, the genetic diversity of P. mugo in Kursiu Nerija is very high and is structured based on the sub-species morphotypes rather than geography. The high frequency of shared and notable frequency of private haplotypes in most of the populations indicate that the major part of the P. mugo material originates from a number of geographically and genetically related sources, which more likely are introductions from abroad that the local collections. The high frequency of private haplotypes in the northernmost populations leaves a possibility for minor introductions from other genetically distinct sources. The absence of private haplotypes in one of the sampled populations indicates the use of local seed collections. The large number of shared haplotypes provides a strong evidence for a geneflow among the P. mugo taxa.     

Semi‐quantitative methods yield greater inter‐ and intraobserver agreement than subjective methods for interpreting 99mtechnetium‐hydroxymethylene‐diphosphonate uptake in equine thoracic processi spinosi

Scintigraphy is a standard diagnostic method for evaluating horses with back pain due to suspected thoracic processus spinosus pathology. Lesion detection is based on subjective or semi‐quantitative assessments of increased uptake. This retrospective, analytical study is aimed to compare semi‐quantitative and subjective methods in the evaluation of scintigraphic images of the processi spinosi in the equine thoracic spine. Scintigraphic images of 20 Warmblood horses, presented for assessment of orthopedic conditions between 2014 and 2016, were included in the study. Randomized, blinded image evaluation was performed by 11 veterinarians using subjective and semi‐quantitative methods. Subjective grading was performed for the analysis of red‐green‐blue and grayscale scintigraphic images, which were presented in full‐size or as masked images. For the semi‐quantitative assessment, observers placed regions of interest over each processus spinosus. The uptake ratio of each processus spinosus in comparison to a reference region of interest was determined. Subsequently, a modified semi‐quantitative calculation was developed whereby only the highest counts‐per‐pixel for a specified number of pixels was processed. Inter‐ and intraobserver agreement was calculated using intraclass correlation coefficients. Inter‐ and intraobserver intraclass correlation coefficients were 41.65% and 71.39%, respectively, for the subjective image assessment. Additionally, a correlation between intraobserver agreement, experience, and grayscale images was identified. The inter‐ and intraobserver agreement was significantly increased when using semi‐quantitative analysis (97.35% and 98.36%, respectively) or the modified semi‐quantitative calculation (98.61% and 98.82%, respectively). The proposed modified semi‐quantitative technique showed a higher inter‐ and intraobserver agreement when compared to other methods, which makes it a useful tool for the analysis of scintigraphic images. The association of the findings from this study with clinical and radiological examinations requires further investigation.