Implication of Lake Water Residence Time on the Classification of Norwegian Surface Water Sites into Progressive Stages of Nitrogen Saturation

Seasonal behaviour of NO₃ ^- in surfacewater is often used as an indicator on a catchment's ability toretain N from atmospheric deposition. In this paper, weclassify 12 pristine sites (five streams and seven lakes) insouthernmost Norway according to the N saturation stageconcept. All examined sites were acid-sensitive and had annualmean NO₃ ^- concentrations in the range 6–36 μeqL^-1. At lake sites with relatively long water residencetimes, hydrology will have a damping effect on the seasonalNO₃ ^- oscillations in the outlet streams. Under givenrunoff conditions (1.0–2.2 m yr^-1), the seasonalNO₃ ^- pattern was clearly affected when lake:catchmentarea ratios exceeded 0.15 and the total residence time of lakesin the catchments exceeded 0.3 yr. In such cases, annualmean NO₃ ^- concentration rather than seasonalvariations may be the better indicator of N saturation. Toaccount for this we propose a set of supplementary criteria forsuch lake systems, defining limit values for annual meanNO₃ ^- concentrations within each stage class. Beforebeing applied on a broader scale, however, we recommend anexamination of additional lake-dominated catchmentsrepresenting a larger gradient in ecosystem types, N depositionlevels and hydrologic regimes. When applying the supplementarycriteria on the Norwegian sites, two were classified as stage 0(no saturation), five as stage 1 (early stage of saturation)and five as stage 2 (saturated – moderate N loss). No siteswere showing symptoms of stage 3, which characteriseswatersheds that are net sources of N, rather than sinks. Theresults indicate a great variability in N retention capacity inthe study region, despite the fact that many of the catchmentsexperience relatively uniform N deposition amounts and climaticconditions. This suggests that much of this variability must bedue to specific catchment characteristics as e.g. soil type,soil depth, and vegetation cover.     

Navicular bone disease: Results of treatment using egg-bar shoeing technique

Although the diagnostic criteria of navicular disease are generally accepted in practice, their limitations are recognised. However, the authors believe that navicular disease per se is reversible and that only secondary changes like adhesions to the deep flexor tendon or spur formations are irreversible. This hypothesis is supported by the results presented of the effect of the application of an egg-bar shoe, which caused permanent cessation of signs of navicular disease in more than 50 per cent of treated cases. The egg-bar shoeing technique can be usefully applied in practice.     

Antimicrobial susceptibility of Haemophilus parasuis and Histophilus somni from pigs and cattle in Denmark

A total of 52 Haemophilus parasuis and 80 Histophilus somni isolates were tested for antimicrobial susceptibility by MIC-determinations. None of the isolates were resistant to ampicillin, ceftiofur, ciprofloxacin, erythromycin, florphenicol, penicillin, spectinomycin, tetracycline, tiamulin, or tilmicosin. Two H. parasuis isolates were resistant to trimethoprim + sulfamethoxazole. Six H. parasuis isolates had reduced susceptibility (0.06-0.5 microg/ml) to ciprofloxacin and 10 reduced susceptibility to TMP + sulfamethoxazole (1-2 microg/ml). This study showed that Danish isolates of H. parasuis and H. somni in general are fully susceptible to antimicrobial agents currently used for treatment of infections with these pathogens.     

Femoral bone adaptation to unstable long-term cemented total hip arthroplasty in dogs

OBJECTIVE: To evaluate femoral adaptation after unstable long-term cemented total hip arthroplasty (cTHA) in dogs. STUDY DESIGN: Clinical study. ANIMALS: Four dogs. METHODS: Paired femurs were examined from client-owned dogs that were donated to a retrieval program after death from causes unrelated to their cTHA. Mean (+/-SEM) dog age was 10.0+/-1.5 years and implant duration was 6.0+/-1.4 years. Implanted femurs had radiographic changes associated with implant loosening and gross mechanical instability at femur retrieval. Femurs were evaluated at 3 levels relative to implant length. Cortical area and medullary area were measured at each level, and cortical porosity was assessed at each level in 4 quadrants and in 3 regions. Implanted femurs were compared with the contralateral non-implanted femurs. RESULTS: Cortical area and cortical porosity were increased at all levels in femurs with unstable implants. Implanted femurs had increased porosity in all quadrants and regions at the proximal 2 levels, and increased porosity in only the cranial quadrant and mid-cortical area at the distal level, when compared with contralateral non-implanted femurs. Corresponding medullary areas were not different. CONCLUSIONS: Significant histomorphometric changes occur in femurs after unstable cTHA. The patterns of periosteal bone formation and endosteal bone resorption support mechanisms of stress shielding and wear debris-mediated osteolysis as factors that may contribute to femoral adaptation and implant loosening. CLINICAL RELEVANCE: Despite popularity and excellent return to function with cTHA, aseptic loosening remains a serious long-term complication. Substantial net bone loss and unfavorable environment with unstable cTHA may make revision surgery less successful.     

Characterization of hemiacetal forms of anthocyanidin 3-O-beta-glycopyranosides

The 3-O-beta-glucopyranosides of delphinidin, petunidin, and malvidin (1-3) and cyanidin 3-O-beta-galactopyranoside (4) dissolved in deuterated methanolic solutions without and with acid (5%, CF3COOD) were identified by homo- and heteronuclear NMR techniques. The hemiacetal forms of all the four anthocyanins were characterized as two epimeric 2-hydroxy-hemiacetals on the basis of assignments of both proton and carbon NMR signals together with chemical shift considerations. This is the first report of 13C NMR assignments of two epimeric anthocyanin hemiacetal forms. No 4-hydroxy-hemiacetal form was detected for any of the pigments. For each anthocyanin dissolved in deuterated methanol, the equilibrium between each of the two epimeric hemiacetals and the corresponding flavylium cation was confirmed by the observed positive exchange cross-peaks in the 2D 1H NOESY spectra. The molar proportions of the flavylium cation and the two hemiacetals of 1-4 in deuterated methanol were very similar for all pigments, even during storage for weeks. The majority of the anthocyanins reported to occur in fruits have the same or similar structures as 1-4. These pigments have been proposed to exist predominantly as hemiacetals in slightly acidic to neutral solvents, which is a relevant pH range in plants and in the human gastrointestinal tract.     

Optimization of conditions for in vitro production of radical oxygen species and expression of tissue factor by canine mononuclear cells and granulocytes for use in high-throughput assays

The purpose of this study was to optimize conditions for high throughput measurement of radical oxygen species (ROS) production and expression of tissue factor, also termed procoagulant activity, by canine leukocytes. Granulocytes and mononuclear cells were separated by density gradient centrifugation from peripheral blood collected on several occasions from three healthy large breed dogs. To determine optimal conditions for ROS production, granulocytes were incubated for 1 or 3h in PBG (PBS containing 0.5% BSA and 5mM glucose) or RPMI containing 10% fetal bovine serum (FBS); lipopolysaccharide (LPS), zymosan, peptidoglycan (PGN) and phorbol myristate acetate (PMA) were used as stimuli. ROS was assessed by conversion of the nonfluorescent dye dihydrorhodamine 123 to fluorescent rhodamine 123 by radical species released into the media. To identify optimal conditions for expression of tissue factor, mononuclear cells were incubated for 5h in RPMI containing different concentrations of heat-inactivated FBS (HI-FBS), and LPS, zymosan, PGN or PMA as stimuli. Expression of tissue factor was determined using a one-stage recalcification assay performed in an automated nephelometric coagulation analyzer. Neither LPS nor zymosan increased ROS production by granulocytes incubated in PBG media. In contrast, granulocytes incubated in RPMI had dose-dependent increases in ROS production in response to zymosan and PGN. ROS production was significantly increased by incubation with concentrations of LPS of 0.01microg/ml or greater, and by zymosan concentrations of 0.1microg/ml or greater. ROS production in response to incubation with PMA was significantly increased starting at 10(-7)M, and was significantly greater for cells incubated in RPMI than cells incubated in PBG. LPS-, zymosan- and PGN-stimulated procoagulant activity increased in a dose-dependent manner, whereas PMA-stimulated procoagulant activity peaked at 10(-7)M. Increasing concentrations of HI-FBS significantly increased LPS-, zymosan- and PGN-induced procoagulant activity of mononuclear cells. Results obtained in this study indicate production of ROS by canine granulocytes is optimal when these cells are incubated for 3h in RPMI with LPS (0.1microg/ml), zymosan (10 microg/ml), PGN (10 microg/ml), and PMA (10(-7)M). Furthermore, canine mononuclear cells express procoagulant activity in response to LPS, zymosan, PGN, and PMA, and responses to LPS, zymosan and PGN are enhanced by the addition of HI-FBS. These findings suggest that HI-FBS retains important serum proteins that facilitate interactions between each of these bacterial or yeast derived products and the mononuclear cells. Consequently, future studies regarding the regulation of procoagulant activity by canine mononuclear cells should be performed in the presence of HI-FBS. Both assays utilized in this study allow high throughput of samples, and therefore are appropriate choices for rapid screening of conditions and/or therapeutic interventions affecting the canine inflammatory system.     

Complications of tibial plateau levelling osteotomy in dogs

The tibial plateau levelling osteotomy (TPLO) is one of the most common surgical procedures used to treat cranial cruciate ligament disease in dogs. Complications occurring during or after TPLO can range in severity from swelling and bruising to fracture and osteomyelitis. Ten to 34% of TPLO surgical procedures are reported to experience a complication and approximately two to four percent require revision surgery to address a complication. Although the risk factors for many complications have not been fully assessed, the best available evidence suggests that complications of TPLO can be reduced with increased surgeon experience, careful surgical planning, and accurate execution of the surgical procedure. Identification of known or suspected risk factors and intra-operative technical errors allow subsequent action to be taken that is aimed at decreasing postoperative morbidity. There is a need for prospective studies with consistent data reporting in order to fully reveal the incidence risk factors for complications associated with TPLO.     

Multiple-locus variant-repeat assay (MLVA) is a useful tool for molecular epidemiologic analysis of Streptococcus agalactiae strains causing bovine mastitis

Group B streptococci (GBS) were considered a major cause of mastitis in cattle until preventive measures succeeded in controlling the disease in the 1970s and 1980s. During the last 5-6 years an increasing number of cases have been observed in some Scandinavian countries. A total of 187 GBS isolates from mastitis cases were collected from 119 animals in 34 Norwegian farms in the period from April 2007 to November 2010. 133 (71%) of the isolates were from farms with automated milking systems. The strains underwent typing of capsular polysaccharides (CPS) and surface proteins, and were analyzed by multi-locus variable repeat assay (MLVA) to investigate the epidemiological relationship of strains within and between farms. The GBS strains were differentiated into 12 types by CPS and surface protein analysis, with CPS types V (54%) and IV (34%) predominating. MLVA was superior to CPS and protein typing for strain differentiation, resolving the 187 strains into 37 types. In 29 of 34 farms all GBS strains had identical MLVA profiles specific for each farm. However, in one farm represented with 48 isolates, four MLVA variants with differences in one repeat locus were observed during the almost 3-year long collection period. Similar variations were observed at four other farms. This might reflect the stability of repeat loci under in vivo conditions. Farms with automated milking systems were overrepresented in this material. In conclusion, the five-loci MLVA allowed rapid high-resolution genotyping of the bovine GBS strains within and between farms.     

Isoenzyme studies on Theileria (Protozoa,Sporozoa). Enzyme activity associated with the erythrocytic stage

Summary

Bovine blood containing piroplasms of Theileria parva, as well as non‐injected blood, was lysed and subjected to iso‐electric focussing.

Staining for 13 different enzymes revealed parasite‐associated bands of glucose phosphate isomerase (GPI) activity, not of any of the other enzymes. There were no variations between individual donor animals in the host cell GPI bands and these bands did not interfere with the recognition of the parasite‐associated bands, so that purification of the piroplasms was unnecessary. Blood from cattle infected with T. mutans also gave parasite‐associated bands of GPI, but no such bands were seen in zymograms of blood from cattle infected with a Theileria sp. from Japan. Depending on the level of parasitaemia, up to four parasite‐associated bands were found in one strain of T. parva and up to three in two other strains. Among the disadvantages of using piroplasm material for the study of isoenzymes of T. parva is the fact that animals often die before their parasitaemia is sufficiently high, and that some strains never give rise to a high parasitaemia.