Interpreting the dependence of soil respiration on soil temperature and water content in a boreal aspen stand

Continuous half-hourly measurements of soil CO₂ efflux made between January and December 2001 in a mature trembling aspen stand located at the southern edge of the boreal forest in Canada were used to investigate the seasonal and diurnal dependence of soil respiration (R_s) on soil temperature (T_s) and water content (θ). Daily mean R_s varied from a minimum of 0.1 μmol m⁻² s⁻¹ in February to a maximum of 9.2 μmol m⁻² s⁻¹ in mid-July. Daily mean T_s at the 2-cm depth was the primary variable accounting for the temporal variation of R_s and no differences between Arrhenius and Q₁₀ response functions were found to describe the seasonal relationship. R_s at 10 °C (R_s10) and the temperature sensitivity of R_s (Q_10Rs) calculated at the seasonal time scale were 3.8 μmol m⁻² s⁻¹ and 3.8, respectively. Temperature normalization of daily mean R_s (R_sN) revealed that θ in the 0–15 cm soil layer was the secondary variable accounting for the temporal variation of R_s during the growing season. Daily R_sN showed two distinctive phases with respect to soil water field capacity in the 0–15 cm layer (θ_fc, 0.30 m³ m⁻³): (1) R_sN was strongly reduced when θ decreased below θ_fc, which reflected a reduction in microbial decomposition, and (2) R_sN slightly decreased when θ increased above θ_fc, which reflected a restriction of CO₂ or O₂ transport in the soil profile.Diurnal variations of half-hourly R_s were usually out of phase with T_s at the 2-cm depth, which resulted in strong diurnal hysteresis between the two variables. Daily nighttime R_s10 and Q_10Rs parameters calculated from half-hourly nighttime measurements of R_s and T_s at the 2-cm depth (when there was steady cooling of the soil) varied greatly during the growing season and ranged from 6.8 to 1.6 μmol m⁻² s⁻¹ and 5.5 to 1.3, respectively. On average, daily nighttime R_s10 (4.5 μmol m⁻² s⁻¹) and Q_10Rs (2.8) were higher and lower, respectively, than the values obtained from the seasonal relationship. Seasonal variations of these daily parameters were highly correlated with variations of θ in the 0–15 cm soil layer, with a tendency of low R_s10 and Q_10Rs values at low θ. Overall, the use of seasonal R_s10 and Q_10Rs parameters led to an overestimation of daily ranges of half-hourly R_s (ΔR_s) during drought conditions, which supported findings that the short-term temperature sensitivity of R_s was lower during periods of low θ. The use of daily nighttime R_s10 and Q_10Rs parameters greatly helped at simulating ΔR_s during these periods but did not improve the estimation of half-hourly R_s throughout the year as it could not account for the diurnal hysteresis effect.     

Comparison of biopsy samples obtained using standard endoscopic instruments and the harmonic scalpel during laparoscopic and laparoscopic-assisted surgery in normal dogs

Objectives— To compare tissue specimens of canine abdominal organs collected by standard techniques (ST) and harmonic scalpel (HS) and to determine the effect of each technique on wound healing.
Study Design— Experimental.
Animals— Twelve dogs.
Methods— Paired biopsy samples (ST, HS) were collected from liver, spleen, adrenal gland, pancreas, stomach, jejunum, and bladder using laparoscopic or laparoscopic-assisted methods. Hemorrhage at biopsy sites was assessed (present or absent). Specimens were evaluated for diagnostic quality and histologic changes. Dogs were necropsied at 4 or 14 days postoperatively.
Results— HS incision caused less frequent hemorrhage of all organs except spleen. HS specimens had more coagulation necrosis than ST specimens, but both techniques yielded specimens that were sized similarly and were of diagnostic quality. At necropsy, HS biopsy sites of the stomach, jejunum, bladder, adrenal gland, and pancreas were more hyperemic than ST sites. There were more adhesions at jejunal and pancreatic ST biopsy sites. HS biopsy sites had more coagulation necrosis at days 4 and 14 postoperatively. Differences in fibrin deposition, inflammation, and fibrosis were present at biopsy sites of some organs at days 4 or 14 and in comparisons between days 4 and 14.
Conclusions— Diagnostic quality biopsy specimens were obtained with HS and ST. Although HS-induced gross and histologic changes during the first 2 postoperative weeks, no clinical complications were observed.
Clinical Relevance— Both HS and ST can yield specimens with minimal hemorrhage and HS resulted in no apparent postoperative problems in normal dogs. Although HS caused more inflammation and adhesions at biopsy sites of the pancreas, adrenal gland, and jejunum, no clinical complications occurred.     

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.     

Development of a two-step nested duplex PCR assay for the rapid detection of Brachyspira pilosicoli and Brachyspira intermedia in chicken faeces

Avian intestinal spirochaetosis (AIS) is an infection of the caeca and/or colo-rectum of laying and meat breeder hens caused by anaerobic intestinal spirochaetes of the genus Brachyspira. AIS can result in a variety of symptoms, including delayed and/or reduced egg production, and increased faecal water content. The two most commonly reported Brachyspira species involved in AIS are Brachyspira pilosicoli and Brachyspira intermedia, and their detection and identification can be difficult and time consuming. In the current study a two-step nested duplex PCR (2S-N-D-PCR) was developed for the detection of these two species, using DNA extracted from washed chicken faeces. In the first step, a duplex PCR (D-PCR) amplifying Brachyspira genus-specific portions of the 16S rRNA and NADH oxidase (nox) genes was undertaken on the washed faeces. In the second step, a nested D-PCR was used that amplified species-specific portions of the 16S rRNA gene of B. pilosicoli and the nox gene of B. intermedia from the amplicons produced in the first step. The 2S-N-D-PCR was rapid and specific, and could be used to detect approximately 10(3) cells of each spirochaete species per gram of washed faeces. When tested on 882 chicken faecal samples from infected flocks, it detected 4-5% more positive faecal samples than did the standard method of selective anaerobic culture followed by individual species-specific PCR assays conducted on the growth on the primary plate. The application of this new technique should improve diagnostic capacity, and facilitate further studies on AIS.     

Effects of intravenous diazepam or microdose medetomidine on propofol-induced sedation in dogs

This crossover study tested the hypothesis that both diazepam and microdose medetomidine would comparably reduce the amount of propofol required to induce sedation. Four different medications, namely high-dose diazepam (0.4 mg/kg intravenously [IV]), low-dose diazepam (0.2 mg/kg IV), medetomidine (1 mug/kg IV), and placebo (0.5 mL physiological saline IV) were followed by propofol (8 mg/kg IV) titrated to a point where intubation could be performed. The effects of medetomidine were comparable to the effects of high-dose diazepam and significantly better than the effects of low-dose diazepam or placebo. Dogs in all treatment groups had transient hypoxemia, and induction and recovery qualities were similar.