Organic‐carbon and nitrogen stocks and organic‐carbon fractions in soil under mixed pine and oak forest stands of different ages in NE Germany

The objective of this work was to evaluate the C and N stocks and organic‐C fractions in soil under mixed forest stands of Scots pine (Pinus sylvestris L.) and Sessile oak (Quercus petraea [Matt.] Liebl.) of different ages in NE Germany. Treatments consisted of pure pine (age 102 y), and pine (age 90–102 y) mixed with 10‐, 35‐, 106‐, and 124‐y‐old oak trees. After sampling O layers, soils in the mineral layer were taken at two different depths (0–10 and 10–20 cm). Oak admixture did not affect total organic‐C (TOC) and N stocks considering the different layers separately. However, when the sum of TOC stocks in the organic and mineral layers was considered, TOC stocks decreased with increasing in oak age (r² = 0.58, p < 0.10). The microbial C (C_MB) was not directly correlated with increase of oak age, however, it was positively related with presence of oak species. There was an increase in the percentage of the C_MB‐to‐TOC ratio with increase of oak‐tree ages. On average, light‐fraction C (C_LF) comprised 68% of the soil TOC in upper layer corresponding to the highest C pool in the upper layer. C_LF and heavy‐fraction C (C_HF) were not directly affected by the admixture of oak trees in both layers. The C_HF accounted on average for 30% and 59% of the TOC at 0–10 and 10–20 cm depths, respectively. Despite low clay contents in the studied soils, the differences in the DCB‐extractable Fe and Al affected the concentrations of the C_HF and TOC in the 10–20 cm layers (p < 0.05). Admixture of oak in pine stands contributed to reduce topsoil C stocks, probably due to higher soil organic matter turnover promoted by higher quality of oak litter.     

Proanthocyanidins: target compounds as antibacterial agents

Grape seeds accumulate in huge quantities as byproduct during wine production and are therefore a cheap source for pharmacologically active agents. However, studies prove poor antibacterial activity, and results of analyses are sometimes contradictory. The aim of this study was, thus, to determine the antibacterial activity of grape seed extracts with special focus on the chromatographic characterization of active fractions. In the course of these investigations, extraction protocols were optimized so that microwave-assisted extraction (MAE) guaranteed highest preconcentration efficiency. Proanthocyanidins, monomeric flavonoid aglycones, as well as some of their glycosides could be identified within yielded extracts via high-performance liquid chromatography-mass spectrometry (HPLC-MS). By that means the coherence number of possible isomers of procyanidins was approximated by a newly developed equation. As far as antibacterial activity determined via screening tests is concerned, the extracts generally have been found to be positively responsive toward 10 different gram-positive and gram-negative bacteria strains. After fractionation of the raw extracts, proanthocyanidins P2, P3, P4 and gallate esters P2G and P3G (P = proanthocyanidin consisting of catechin and epicatechin units, n = oligomerization degree, G = gallate ester) were determined as active antibacterial agents toward 10 different pathogens. Only moderate activity was found for monomeric flavonoid fractions.     

Screening for nutritive peptides that modify cholesterol 7alpha-hydroxylase expression

Bioactive peptides with a variety of effects have been described from several nutritive proteins. They exhibit antimicrobial, blood-pressure lowering, antithrombotic, immunomodulatory, and cholesterol-modulating effects. In this study, we have examined whether peptides derived from food proteins might influence bile acid synthesis. A reporter gene cell line that carries a cholesterol 7alpha-hydroxylase promoter fragment fused to firefly luciferase ( cyp7a-luc) was used to screen for nutritive peptides affecting cyp7a expression, the enzyme catalyzing the rate-limiting step in bile acid synthesis. Proteolytic hydrolysates were prepared from soy protein and bovine casein with pepsin, trypsin, chymotrypsin, and elastase and size fractionated using ultrafiltration. Several bioactive hydrolysates could be identified that inhibited luciferase expression. Also, an activation of kinase (AKT, ERK, p38-MAPK) signaling could be observed. Selected hydrolysates were further fractionated by reversed-phase HPLC. Bioactive HPLC-fractions were obtained from casein but not from soy hydrolysates; however, activity could not be recovered in single peak fractions. Peptides in such fractions were identified by mass spectrometry. Five selected peptides from alpha S1-casein present in active fractions were synthesized, but none of these showed activity in the cyp7a-luc screening system. However, two of them activated MAP-kinase signaling similar to the hydrolysates, which suggests, that these peptides are involved in cyp7a regulation by the casein hydrolysates.     

Haemostatic abnormalities in cats with naturally occurring liver diseases

Alterations in the haemostatic system were characterized in cats with different naturally occurring liver diseases. The study looked at 44 healthy cats and 45 cats with different liver diseases confirmed histologically or cytologically (neoplasia, n=9; inflammation, n=12; hepatic lipidosis, n=13; other degenerative liver diseases, n=11). The following parameters were evaluated: platelet count; prothrombin time; activated partial thromboplastin time; thrombin time; factor (F) II, FV, FVII, FX, and FXIII activities; fibrinogen concentration; activities of antithrombin, protein C, plasminogen, and α(2)-plasmin inhibitor, and D-dimer concentration. In cats with liver diseases, 44/45 (98%) had one or more abnormalities of the coagulation parameters measured. In cats with inflammatory liver diseases, increased D-dimer concentrations and decreased FXIII activity were the most consistent abnormalities and were found in 83% and 75% of cats, respectively. The most common abnormality in cats with neoplastic liver disease was FXIII deficiency (78%). The most consistent abnormalities in cats with hepatic lipidosis were increased FV activity and D-dimer concentration with 54% of cats having values above the reference range for both parameters. Cats with miscellaneous degenerative liver disease most frequently showed FXIII deficiency (64%). The results of this study show that alterations of single haemostatic components are a frequent finding in cats with liver disease. Activation of haemostasis with subsequent consumptive coagulopathy (rather than decreased synthesis) seems to be responsible for these alterations. Increased blood levels of different haemostatic components in cats with inflammatory lesions may be related to an acute phase reaction.     

Histological, histochemical and immunohistochemical study of the lymph nodes of the one humped camel (Camelus dromedarius)

Prescapular, femoral, mesenteric, mediastinal and splenic lymph nodes from nine camels of one to 12 years of age were studied. There were no obvious structural differences between these different lymph nodes or between the ages. The lymph nodes were surrounded by a capsule formed of two layers, an outer thicker layer of connective tissue and an inner thinner layer mainly of smooth muscles. Trabeculae extended from the inner layer of the capsule dividing the parenchyma characteristically into incomplete lobules. Subcapsular and trabecular lymphatic sinuses were supported by a reticular fiber network. The parenchyma was uniquely different from that of other species, as it was arranged in the form of lymphoid follicles and interfollicular lymphoid tissue. The lymphoid follicles of CD22 positive lymphocytes were supported by a reticular fiber network. This fine network of α-smooth muscle actin positive cells enclosed the lymphoid follicles. The interfollicular tissue was mainly made up of diffusely distributed CD3 positive lymphocytes. MHC class II: DR was expressed by most lymphocytes of the follicles and interfollicular tissue. Lymphatic sinuses and high endothelial venules were found in the interfollicular zone. The lymphatic sinuses were lined by discontinuous endothelial cells. The wall of the high endothelial venules was infiltrated by several lymphocytes and enclosed in a layer of α-smooth muscle actin positive cells. Acid phosphatase positive cells were evenly distributed in the interfollicular zone. A few cells were localized in the lymph follicles. Alkaline phosphatase was observed in the endothelium of the lymphatic sinuses and in the lymphoid follicles.     

Corynebacterium ulcerans infection in roe deer (Capreolus capreolus)

This is the first report of a Corynebacterium (C) ulcerans-infection in European roe deer (Capreolus capreolus). The bacterium was isolated from a grapefruit sized abscess of an animal that had been shot. In addition to biochemical tests, the isolate was identified by Fourier transform infrared spectroscopy (FT-IR) and partial sequencing of the rpoB gene. The isolated bacteria showed phospholipase D activity that could be demonstrated by reverse CAMP-test. A tox-gene could be detected by PCR but the Elek-test specific for diphtheria toxin failed.The isolate was compared to two C. ulcerans-strains isolated from wild boar (Sus scrofa) from the state of Baden-Wuerttemberg described recently.     

Evaluation of variables to describe the shape of volumetric capnography curves during bronchoconstriction in dogs

The aim of the study was to investigate changes in volumetric capnography (V(C)) variables during bronchoconstriction in dogs and compare it with total respiratory resistance (R(L)) measured with a Fleisch pneumotachograph. Six dogs were challenged with increasing concentrations of carbachol until obvious signs of bronchoconstriction were seen. All V(C) parameters were obtained before, directly after, 10 and 20 min after maximal bronchoconstriction. The slope of phase III (SIII) and airway and alveolar dead space parameters were significantly different from baseline directly after the challenge. The V(C) curve obtained a typical shape at the time of maximal bronchoconstriction and a trend to return to baseline shape was seen over time. A significant correlation was found for all aforementioned parameters with R(L). We conclude that the shape of the V(C) curve in combination with dead space calculation can be used to verify bronchoconstriction on a breath-to-breath basis.     

PCR-based genetic evidence for occurrence of Helicobacter pylori and novel Helicobacter species in the canine gastric mucosa

The canine gastric mucosa is known to be a habitat for various Helicobacter species. So far, five Helicobacter species have been described from the canine gastric mucosa, but histological studies have demonstrated a greater variety. In order to gain more information on diversity of canine gastric mucosa colonising helicobacters, biopsy samples of four pet dogs were examined by DNA-based techniques. PCR with a primer pair binding specifically to the 16S rDNA of the species of the genus Helicobacter and generating a fragment of approximately 400 bp indicated the presence of Helicobacter strains in the stomachs of the four dogs. PCR products were cloned into Escherichia coli DH10B and PCR-re-amplified 16S rDNA fragments were subjected to amplified ribosomal DNA restriction analysis (ARDRA) employing restriction enzyme HhaI. Restriction profiles indicated the presence of at least two different Helicobacter species in two dogs. Partial sequences of 16S rDNA of six clones were compared with sequences available in the EMBL data bank. Two sequences obtained from different dogs were identical with the corresponding sequences of Helicobacter pylori strains. Three sequences showed highest but moderate similarity values to H. pylori (96.6-98.0%) and one sequence to Helicobacter salomonis (97.3%). In contrast to previous reports our data implicate that the gastric mucosa of dogs may be colonised by strains of H. pylori or a very closely related species but they also confirm indications for the presence of so far uncultivated species of Helicobacter.     

Relationship of age,sex, body weight,and hematologic and respiratory variables with airway reactivity in adult cats

OBJECTIVE: To evaluate the association between airway reactivity and age, sex, body weight, and radiographic findings in cats. ANIMALS: 32 mature cats that constituted 2 age groups (17 young cats that were 1 to 2 years old and 15 old cats that were 12 to 13 years old). PROCEDURE: Cats were placed in the chamber of a barometric whole-body plethysmograph (volume, 38 L), and box pressure was measured at baseline and after aerosol administration of increasing concentrations of carbachol. Airway reactivity was assessed by monitoring increases in enhanced pause (PENH), a unitless variable that measures bronchoconstriction as derived from dose-response curves. The endpoint chosen was the provocative concentration of carbachol that increased PENH to 300% of the baseline value (PCPENH300). RESULTS: We did not find a correlation between PCPENH300 and sex, body weight, number of eosinophils, PENH before bronchoconstriction, respiratory frequency, tidal volume, or minute ventilation. Airway reactivity was significantly less in the old cats (mean +/- SD PCPENH300, 0.578 +/- 0.051%), compared with the value for the young cats (0.053 +/- 0.006%). Radiographic patterns differed significantly between groups of cats; a greater proportion of old cats (12/15) had bronchointerstitial patterns, compared with the proportion of young cats (4/17). CONCLUSIONS AND CLINICAL RELEVANCE: These data support the notion that age exerts a strong influence on airway reactivity in adult cats, and radiographic differences suggest that structural changes in older cats may contribute to this effect. These findings have important implications for interpretation of results of airway reactivity tests in cats.