Linoleic acid increases adhesion,chemotaxis, granule release,intracellular calcium mobilisation,MAPK phosphorylation and gene expression in bovine neutrophils

Neutrophils are critical to the innate immune response; therefore, the proper function of neutrophils is critical to avoid the development of certain diseases. Linoleic acid, a polyunsaturated long-chain fatty acid, is one of the most abundant long-chain fatty acids found in the plasma of cows after giving birth. In this study, we evaluated the effects of linoleic acid treatment on bovine neutrophil adhesion, chemotaxis, metalloproteinase (MMP)-9 release, CD11b expression, intracellular calcium mobilisation, mitogen-activating protein kinase (MAPK) phosphorylation and COX-2 and IL-8 expression. Bovine neutrophils isolated from healthy heifers were incubated with different concentrations of linoleic acid, and then neutrophil responses were evaluated. Our results show that the treatment of neutrophils with 100 μM linoleic acid increased their adhesion to the bovine endothelial cell line CPA47. The results of a transwell migration assay revealed that linoleic acid could also promote the chemotaxis of bovine neutrophils. Furthermore, linoleic acid treatment increased MMP-9 activity and CD11b cell surface expression in neutrophils. Fifty and 100 μM linoleic acid also increased intracellular calcium mobilisation in neutrophils loaded with Fluo-4 AM dye. Linoleic acid also rapidly (2–5 min) stimulated the phosphorylation of ERK1/2 and p38 MAPK as evaluated by immunoblot. Finally, COX-2 and IL-8 mRNA expression increased after 2 h of linoleic acid treatment. In conclusion, linoleic acid stimulates adhesion, chemotaxis, granule release and intracellular responses in bovine neutrophils.     

2-Aminoethoxydiphenyl borate (2-APB) reduces alkaline phosphatase release, CD63 expression, F-actin polymerization and chemotaxis without affecting the phagocytosis activity in bovine neutrophils

2-Aminoethoxydiphenyl borate (2-APB) interferes with the Ca(2+) influx and reduces the ROS production, gelatinase secretion and CD11b expression in bovine neutrophils. Moreover, it has been suggested that inhibition of the Ca(2+) channel involved in the store operated Ca(2+) entry (SOCE) is a potential target for the development of new anti-inflammatory drugs in cattle, however it is unknown whether 2-APB affects neutrophil functions associated with the innate immune response. This study describes the effect of 2-APB, a putative SOCE inhibitor, on alkaline phosphatase activity a marker of secretory vesicles, CD63 a marker for azurophil granules, F-actin polymerization and in vitro chemotaxis in bovine neutrophils stimulated with platelet-activating factor (PAF). Also, we evaluated the effect of 2-APB in the phagocytic activity against Escherichia coli and Staphylococcus aureus bioparticles. We observed that doses of 2-APB ≥10 μM significantly reduced alkaline phosphatase activity and in vitro chemotaxis, whereas concentrations of 2-APB ≥50 μM reduced CD63 expression and F-actin polymerization. Finally, we observed that 2-APB did not affect the phagocytic activity in neutrophils incubated with E. coli and S. aureus bioparticles. We concluded that inhibition of Ca(2+) influx could be a useful strategy to reduce inflammatory process in cattle.     

D-lactic acid interferes with the effects of platelet activating factor on bovine neutrophils

D-lactic acidosis occurs in ruminants, such as cattle, with acute ruminal acidosis caused by ingestion of excessive amounts of highly fermentable carbohydrates. Affected animals show clinical signs similar to those of septic shock, as well as acute laminitis and liver abscesses. It has been proposed that the inflammatory response and susceptibility to infection could both be caused by the inhibition of phagocytic mechanisms. To determine the effects of d-lactic acid on bovine neutrophil functions, we pretreated cells with different concentrations of D-lactic acid and measured intracellular pH using 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) and calcium flux using FLUO-3 AM-loaded neutrophils. Reactive oxygen species (ROS) production was measured using a luminol chemiluminescence assay, and MMP-9/gelatinase-B granule release was measured by zymography. CD11b and CD62L/l-selectin expression, changes in cell shape, superoxide anion production, phagocytosis of Escherichia coli-Texas red bioparticles, and apoptosis were all measured using flow cytometry. Our results demonstrated that D-lactic acid reduced ROS production, CD11b upregulation and MMP-9 release in bovine neutrophils treated with 100 nM platelet-activating factor (PAF). D-lactic acid induced MMP-9 release and, at higher concentrations, upregulated CD11b expression, decrease L-selectin expression, and induces late apoptosis. We concluded that D-lactic acid can interfere with neutrophil functions induced by PAF, leading to reduced innate immune responses during bacterial infections. Moreover, the increase of MMP-9 release and CD11b expression induced by 10mM D-lactic acid could promote an nonspecific neutrophil-dependent inflammatory reaction in cattle with acute ruminal acidosis.     

Improvements of immune status, intestinal integrity and gain performance in the early-weaned calves parenterally supplemented with L-alanyl-L-glutamine dipeptide

Glutamine (Gln) is an important substrate for the innate immune cells including lymphocytes and macrophages. In this report, the effects of alanyl-glutamine dipeptide (Ala-Gln) on the naïve immune system, intestinal integrity and gain performance of early-weaned calves were investigated. Early-weaned Chinese Holstein calves were intravenously administered different dosages of Ala-Gln. The effects of Ala-Gln administration were ascertained by evaluating the blood for naïve T lymphocyte subpopulations, the concentrations of serum IgG, serum IgA and intestinal mucosal secretory IgA (s-IgA), the intestinal integrity, as well as the gain performance. Results demonstrated that intravenous administration of Ala-Gln dipeptide (1.01 g/kg × d^?1) for 7 days had a positive effect on gain performance, intestinal integrity and the immune system. Calves administered doses of Ala-Gln displayed an improvement in gain performance and health status concurrent with increases in blood CD2⁺ and CD4⁺ lymphocytes, the ratio of CD4⁺/CD8⁺, serum IgA and IgG, intestinal mucosal s-IgA while decreasing the occurrence of diarrhea. Moreover, we found that animals given the effective dose (1.01 g/kg × d^?1) of Ala-Gln resulted in improved immune status and intestinal integrity relative to those given a lower (0.49 g/kg × d^?1) or higher dose (1.99 g/kg × d^?1) of Ala-Gln. These findings suggest that maintaining a certain concentration of plasma and/or tissue glutamine in the early stages of weaning is an effective alternative approach for improvement of growth performance in early-wean calves.     

Advanced oxidation protein products are generated by bovine neutrophils and inhibit free radical production in vitro

Despite the recognised importance of oxidative stress in the health and immune function of dairy cows, protein oxidation markers have been poorly studied in this species. The current study aimed to characterise markers of protein oxidation generated by activated bovine neutrophils and investigate the biological effects of advanced oxidation protein products (AOPP) on bovine neutrophils. Markers of protein oxidation (AOPP, dityrosines and carbonyls) were measured in culture medium containing bovine serum albumin (BSA) exposed to neutrophils. The effect of AOPP-BSA on generation of reactive oxygen species (ROS) was assessed by chemiluminescence. Activation of caspases-3, -8 and -9 and the presence of DNA laddering were used as apoptosis markers.Greater amounts of AOPP were generated by phorbol myristate acetate (PMA)-activated than non-activated neutrophils (1.46 ± 0.13 vs. 0.75 ± 0.13 nmol/mg protein, respectively; P < 0.05). Activated neutrophils and hypochlorous acid generated slightly different patterns of oxidized protein markers. Exposure to AOPP-BSA did not stimulate ROS production. Activated neutrophils generated a lesser amount of ROS when incubated with AOPP-BSA (P < 0.001). Activation with PMA induced a loss of viable neutrophils after 3 h, which was greater with AOPP-BSA incubation (P < 0.05). Detectable amounts of active caspases-3, -8 and -9 were found in nearly all samples but differences in caspase activation or DNA laddering were not observed comparing treatment groups. Apoptosis was unlikely to be responsible for the greater loss of PMA-activated neutrophils cultured in AOPP-BSA and it is possible that primary necrosis occurred. The results suggest that accumulation of oxidized proteins at an inflammatory site might result in a progressive reduction of neutrophil viability.     

The impact of MK-467 on sedation,heart rate and arterial blood pressure after intramuscular coadministration with dexmedetomidine in conscious cats

Objective

To study the effects of MK-467, a peripheral α₂-adrenoceptor antagonist, on sedation, heart rate and blood pressure after intramuscular (IM) coadministration with 25 μg kg^?1 of dexmedetomidine in cats.

Retinoic acid induces apoptosis in activated canine neutrophils

Activated neutrophils live longer, produce toxic metabolites and cause considerable tissue injury, which is central to the pathogenesis of many inflammatory conditions. Retinoids are a class of lipophilic compounds with anti-inflammatory effects. We examined the effect of retinoic acid on apoptosis in normal and activated neutrophils. Our results showed that treatment with 1 μg/ml Escherichia coli lipopolysaccharide (LPS) for 12 and 36 h delayed the spontaneous neutrophil apoptosis compared to untreated cells. But exposure of LPS-treated cells to retinoic acid (1 and 5 μM) abolished the inhibitory effects of LPS on neutrophil apoptosis in a concentration-dependant manner based on annexin V staining, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, light and electron microscopy. These results show that retinoic acid increases apoptosis in activated canine neutrophils and this effect could enhance the resolution of inflammation in vivo.     

Characterization of Mannheimia haemolytica biofilm formation in vitro

Mannheimia haemolytica is the primary bacterial agent in the bovine respiratory disease complex. It is thought that M. haemolytica colonizes the tonsillar crypts of cattle as a commensal and subsequently descends into the lungs to cause disease. Many bacterial species persist in the host as biofilms. There is limited information about the ability of M. haemolytica to form biofilms. The aim of this study was to develop an in vitro model for M. haemolytica biofilm formation. We found that M. haemolytica required at least 36 h to form robust biofilms on plastic in vitro when incubated in RPMI-1640 tissue culture medium at 37 °C, with maximal biofilm formation being evident at 48 h. Biofilm formation was inhibited by adding the monosaccharides d(+) galactose and d(+) mannose to the growth medium. Addition of antibodies to the M. haemolytica surface protein OmpA also reduced biofilm formation. Upon evaluating the macromolecules within the biofilm extracellular polymeric substance we found it contained 9.7 μg/cm² of protein, 0.81 μg/cm² of total carbohydrate, and 0.47 μg/cm² of extracellular DNA. Furthermore, proteinase K treatment significantly decreased biofilms (P < 0.05) while α-amylase and micrococcal nuclease decreased biofilms to a lesser extent. M. haemolytica biofilm cells were more resistant than planktonic cells to the antibiotics florfenicol, gentamicin, and tulathromycin. These results provide evidence that M. haemolytica can form biofilms, which could contribute to its ability to persist as a commensal in the bovine upper respiratory tract.     

Plasma concentrations of buprenorphine after epidural administration in conscious cats

Buprenorphine plasma concentrations were measured after administering buprenorphine (20 μg/kg) into the lumbosacral epidural space of conscious cats chronically instrumented with an epidural catheter. Blood was collected from a jugular vein before injection and 15, 30, 45 and 60 min and 2, 3, 4, 5, 6, 8, 12 and 24 h after administration. Plasma buprenorphine concentrations were measured using ELISA. Background concentration (before injection) was 1.27 ± 0.27 ng/mL (mean ± SD). Including background concentration, the mean peak plasma concentration was obtained 15 min after injection (5.82 ± 3.75 ng/mL), and ranged from 3.79 to 2.20 ng/mL (30 min–3 h), remaining between 1.93 and 1.77 ng/mL (4–12 h), and declined to 1.40 ± 0.62 ng/mL at 24 h. Elimination half-life was 58.8 ± 40.2 min and clearance 56.7 ± 21.5 mL/min. Results indicate early rapid systemic uptake of buprenorphine from epidural administration with plasma concentrations similar to using buccal or IM routes by 15 min postinjection.     

Simultaneous detection of the feline lungworms Troglostrongylus brevior and Aelurostrongylus abstrusus by a newly developed duplex-PCR

In addition to Aelurostrongylus abstrusus (Strongylida: Angiostrongylidae), referred to as the feline lungworm, Troglostrongylus brevior (Strongylida: Crenosomatidae) has recently been identified as an agent of bronco-pulmonary infestations in cats. These two parasites have a similar biology, share ecological niches, potentially co-infesting cats, but are difficult to be differentiated due to the morphological similarities of their first-stage larvae (L1). This paper describes a molecular tool, based on single-step duplex polymerase chain reaction (duplex-PCR) on the ribosomal internal transcribed spacer 2 region (ITS-2) for the simultaneous detection and differentiation of T. brevior and A. abstrusus. L1 of both species were collected from faecal samples, morphologically identified, and single larval specimens isolated. An aliquot of faeces was used as a test sample for a case of mixed natural infestation. The duplex-PCR was performed using species-specific forward primer sets for the ITS-2 region (i.e., A. abstrusus: 220 bp; T. brevior: 370 bp). The detection limit of the molecular assay was also assessed by serial dilutions of DNA from single larvae of both species (from ～4.0 to 4.0 × 10^?5 μg/μl). The duplex-PCR carried out on individual DNA samples was able to detect as low as 5.2 × 10^?3 μg/μl of DNA for A. abstrusus, 4.9 × 10^?3 μg/μl for T. brevior, and as low as 4.0 × 10^?3 μg/μl for samples containing both species. Species-specific bands of the expected sizes and two bands were simultaneously amplified from the faecal sample containing both species. The phylogenetic analyses of the ITS-2 sequences here examined and those available for other metastrongyloids were concordant in clustering them with those of other Troglostrongylus brevior and A. abstrusus sequences available in GenBank database. This molecular approach proved to be effective and highly sensitive for the simultaneous detection of the two lungworms species and it might be used for molecular epidemiological studies and for monitoring therapeutic protocols.     

Pharmacokinetics,plasma protein binding and bioavailability of moxifloxacin in Muscovy ducks after different routes of administration

In this study the disposition kinetics and plasma availability of moxifloxacin in Muscovy ducks after single intravenous (i.v.), intramuscular (i.m.) and oral (p.o.) administrations of 5 mg kg^?1 b.wt. were investigated. The concentrations of moxifloxacin in the plasma were measured using high-performance liquid chromatography (HPLC) with fluorescence detection on samples collected at frequent intervals after drug administration. Following intravenous injection, the decline in plasma drug concentration was bi-exponential with half-lives of (t_1/2α) 0.22 ± 0.10 h and (t_1/2β) 2.49 ± 0.26 h for distribution and elimination phases, respectively. The volume of distribution at steady-state (V_dss) was 1.02 ± 0.14 l kg^?1 and the total body clearance (Cl_tot) was 0.32 ± 0.11 l kg^?1 h^?1, respectively. After intramuscular and oral administration of moxifloxacin at the same dose the peak plasma concentrations (C_max) were 2.38 ± 0.43 and 2.11 ± 0.36 μg ml^?1 and were obtained at 1.47 ± 0.26 and 1.83 ± 0.16 h (T_max), respectively, the elimination half-lives (T_1/2el) were 3.14 ± 0.42 and 2.63 ± 0.44 h, respectively, and AUC_0–24 were 15.87 ± 2.35 and 14.52 ± 2.37 μg ml^?1 h^?1, respectively. The systemic bioavailabilities were 96.36 ± 11.54% and 86.79 ± 12.64%, respectively. In vitro plasma protein binding percent was 32%. We concluded that moxifloxacin might be clinically interesting alternative for the treatment of most sensitive bacterial infections in Muscovy ducks.     

Marked MMP-2 transcriptional up-regulation in mononuclear leukocytes invading the subarachnoidal space in aseptic suppurative Steroid-Responsive Meningitis–Arteritis in dogs

Canine Steroid-Responsive Meningitis–Arteritis (SRMA) is a suitable animal model for studies on the development of neutrophilic pleocytosis in aseptic meningitis. Samples of dogs in the acute phase of SRMA (n = 16) were examined for gene expression of matrix metalloproteinases (MMP)-2 and -9 and tissue inhibitors of metalloproteinases (TIMP)-1 and -2. Results were compared to those of dogs under glucocorticosteroid treatment for SRMA (n = 16) and dogs with other inflammatory and neoplastic diseases of the central nervous system (CNS) (n = 19). Samples included mononuclear (PBMCs) and polymorphonuclear cells (PBPMNs) of peripheral blood and cerebrospinal fluid white blood cells (CSF WBCs). In the acute phase of SRMA CSF WBCs showed mRNA expression for MMP-2 and -9 and TIMP-1 and -2, highlighting a contribution of these cells to the overall content of MMPs and TIMPs in CSF. MMP-2 mRNA levels in CSF WBCs were significantly up-regulated in comparison to PBMC expression levels, suggesting that MMP-2 is relevant for PBMC invasion into the subarachnoidal space and that the expression is influenced by migratory activity through the blood–CSF-barrier.     

Canine heartworm (Dirofilaria immitis) in Fresno and Madera Counties, California: prevalence differences between foothill and valley habitats

The aim of this study was to assess the prevalence of heartworm in domestic dogs in Madera and Fresno Counties, California, dependent on habitat and other host and environmental factors. Dogs were screened for presence of heartworm antigen using the PetChek^® ELISA on blood samples (N = 519) collected at seven sites during April-July 2009. Eighteen dogs were heartworm antigen positive. Pearson Chi-square analyses were conducted testing the presence of heartworm antigen against the following independent variables: elevation range, percentage of time spent outdoors during the day, percentage of time spent outdoors during the night, pet coat length, weight class, prevention status, and sex. Dogs that spent at least 50% of their time outdoors during the day were significantly more likely to have heartworm that those who spent less time outside (N = 519, df = 1, p = 0.031). Overall prevalence (3.47%) was lower than expected, with Madera County having 3.8% positive samples and Fresno County 3.5%; this prevalence is lower than in many previous studies. The effect of time spent outdoors on heartworm prevalence was similar to previous studies. The impact of elevation on infection, though not significant, requires further investigation, as does the prevalence and viability of larval stages in mosquitoes.     

Vegetation,Hydrologic, and Erosion Responses of Sagebrush Steppe 9 Yr Following Mechanical Tree Removal

Land managers across the western United States are faced with selecting and applying tree-removal treatments on pinyon (Pinus spp.) and juniper (Juniperus spp.) woodland-encroached sagebrush (Artemisia spp.) rangelands, but current understanding of long-term vegetation and hydrological responses of sagebrush sites to tree removal is inadequate for guiding management. This study applied a suite of vegetation and soil measures (0.5 ? 990 m²), small-plot rainfall simulations (0.5 m²), and overland flow experiments (9 m²) to quantify the effects of mechanical tree removal (tree cutting and mastication) on vegetation, runoff, and erosion at two mid- to late-succession woodland-encroached sagebrush sites in the Great Basin, United States, 9 yr after treatment. Low amounts of hillslope-scale shrub (3 ? 15%) and grass (7 ? 12%) canopy cover and extensive intercanopy (area between tree canopies) bare ground (69 ? 88% bare, 75% of area) in untreated areas at both sites facilitated high levels of runoff and sediment from high-intensity (102 mm ? h^? 1, 45 min) rainfall simulations in interspaces (~ 45 mm runoff, 59 ? 381 g ? m^? 2 sediment) between trees and shrubs and from concentrated overland flow experiments (15, 30, and 45 L ? min^? 1, 8 min each) in the intercanopy (371 ? 501 L runoff, 2 342 ? 3 015 g sediment). Tree cutting increased hillslope-scale density of sagebrush by 5% and perennial grass cover by twofold at one site while tree cutting and mastication increased hillslope-scale sagebrush density by 36% and 16%, respectively, and perennial grass cover by threefold at a second more-degraded (initially more sparsely vegetated) site over nine growing seasons. Cover of cheatgrass (Bromus tectorum L.) was < 1% at the sites pretreatment and 1 ? 7% 9 yr after treatment. Bare ground remained high across both sites 9 yr after tree removal and was reduced by treatments solely at the more degraded site. Increases in hillslope-scale vegetation following tree removal had limited impact on runoff and erosion for rainfall simulations and concentrated flow experiments at both sites due to persistent high bare ground. The one exception was reduced runoff and erosion within the cut treatments for intercanopy plots with cut-downed-trees. The cut-downed-trees provided ample litter cover and tree debris at the ground surface to reduce the amount and erosive energy of concentrated overland flow. Trends in hillslope-scale vegetation responses to tree removal in this study demonstrate the effectiveness of mechanical treatments to reestablish sagebrush steppe vegetation without increasing cheatgrass for mid- to late-succession woodland-encroached sites along the warm-dry to cool-moist soil temperature ? moisture threshold in the Great Basin. Our results indicate improved hydrologic function through sagebrush steppe vegetation recruitment after mechanical tree removal on mid- to late-succession woodlands can require more than 9 yr. We anticipate intercanopy runoff and erosion rates will decrease over time at both sites as shrub and grass cover continue to increase, but follow-up tree removal will be needed to prevent pinyon and juniper recolonization. The low intercanopy runoff and erosion measured underneath isolated cut-downed-trees in this study clearly demonstrate that tree debris following mechanical treatments can effectively limit microsite-scale runoff and erosion over time where tree debris settles in good contact with the soil surface.     

Leafy Spurge Suppression by Flea Beetles in The Little Missouri Drainage Basin,USA

The Ecological Area-wide Management Leafy Spurge, or TEAM Leafy Spurge, began collecting and redistributing flea beetles (Aphthona spp.) to research/demonstration sites and landowners throughout the Little Missouri River drainage basin to control leafy spurge in 1998. A study to evaluate the change over time of leafy spurge (Euphorbia esula L.) phytosociological characteristics following release of flea beetles was initiated in 2002 on leafy spurge–infested pasture and rangeland in the Little Missouri River drainage of Montana, North Dakota, South Dakota, and Wyoming. A total of 292 flea beetle release sites were analyzed in June and July 2002 and 2003 for leafy spurge stem density, foliar cover, flea beetle density, and vegetation composition. Leafy spurge stem density suppression was evident at 91% of the study sites. On two-thirds of the study sites stem density was reduced from greater than 100 stems·m⁻¹ to less than 25 stems·m⁻¹. Leafy spurge foliar cover was less than 5% on approximately two-thirds of the flea beetle release sites and less than 25% on over 90% of the release sites. Area of observed leafy spurge suppression ranged from 0 m² to 30000 m². Approximately 40% of the release sites had leafy spurge suppression ranging from 1000 m² to 5000 m², and 14% of the release sites had greater than 10000 m² of leafy spurge control. Plant community composition following leafy spurge suppression was characteristic of native plant communities that had not been burned or grazed. Flea beetles effectively reduced leafy spurge stem density and cover in 4–5 yr across a variety of locations and corresponding environmental conditions, both within the Little Missouri River drainage and in selected nearby locations.     

Radial strain and strain rate by two-dimensional speckle tracking echocardiography and the tissue velocity based technique in the dog

ObjectivesTwo-dimensional (2D) speckle tracking echocardiography (STE) is a new angle-independent ultrasound technique based on tracking of speckles within the myocardium on 2D grayscale images. The aims of this prospective study were as follows: (1) to assess the variability of left ventricular peak systolic radial strain (St) and strain rate (SR) in awake dogs using STE (Protocol 1); and (2) to quantify these variables in a healthy canine population and compare them with tissue Doppler imaging (TDI)-based St and SR values (Protocol 2).BackgroundSt and SR may be assessed using TDI, which is limited by angle dependency.Animals, materials and methodsThirty-six STE examinations were performed on 6 healthy dogs for Protocol 1 and 37 healthy dogs were recruited for Protocol 2. In both studies, STE measurements were obtained offline from the right parasternal short-axis view by the same trained observer using automatic frame-to-frame tracking of grayscale speckle patterns.ResultsAll within- and between-day coefficients of variation were <10% (Protocol 1). In Protocol 2, St (46.7 ± 12.2%) and SR (2.7 ± 0.6 s⁻¹) measured by STE were correlated with heart rate (p < 0.01), but not with the ratio of early mitral inflow velocity to early mitral annular velocity. There was a good correlation between STE and TDI for both St and SR values (p < 0.001).ConclusionsSTE is a repeatable and reproducible non-Doppler method for assessing radial St and SR. The combination of these indices with conventional echo-Doppler variables could provide a new approach for accurately quantifying canine systolic function.     

Evaluation of peripheral lymphocytes after weaning and vaccination for Mycoplasma hyopneumoniae

This study evaluated immune cell populations in pigs following weaning and vaccination for Mycoplasma hyopneumoniae. Piglets (n = 24) were weaned (day 0) at 16 (±1) days of age, and randomly assigned to the vaccination group (n = 16) or control group (n = 8). Complete blood cell counts, flow cytometry and serology were completed for blood samples collected on days 0 (within hours of weaning), 3, 7, 14, 30 and 60. The M. hyopneumoniae S:P ratios (sample optical density: positive control optical density) were negative in the vaccination group until days 30 and 60, when the S:P ratios were 1.3 and 1.0, respectively. Control animals remained serologically negative. The percentage of CD4⁺ T cells was less (P < 0.01) in control pigs than vaccinated pigs at day 3. In contrast, numbers of CD8⁺ and CD4⁺CD8⁺ T cells were greater (P < 0.01) in control pigs than in vaccinated pigs at days 3 and 7. After day 7, few differences in immune cell types were evident between the groups. Differences in lymphocyte populations could not be solely attributed to vaccination, due at least in part, to the confounding influence of weaning. It was difficult to distinguish the influence of vaccination from the impact of weaning on peripheral immune cell populations.     

Oxidative response of neutrophils to platelet-activating factor is altered during acute ruminal acidosis induced by oligofructose in heifers

Reactive oxygen species (ROS) production is one of the main mechanisms used to kill microbes during innate immune response. D-lactic acid, which is augmented during acute ruminal acidosis, reduces platelet activating factor (PAF)-induced ROS production and L-selectin shedding in bovine neutrophils in vitro. This study was conducted to investigate whether acute ruminal acidosis induced by acute oligofructose overload in heifers interferes with ROS production and L-selectin shedding in blood neutrophils. Blood neutrophils and plasma were obtained by jugular venipuncture, while ruminal samples were collected using rumenocentesis. Lactic acid from plasma and ruminal samples was measured by HPLC. PAF-induced ROS production and L-selectin shedding were measured in vitro in bovine neutrophils by a luminol chemiluminescence assay and flow cytometry, respectively. A significant increase in ruminal and plasma lactic acid was recorded in these animals. Specifically, a decrease in PAF-induced ROS production was observed 8 h after oligofructose overload, and this was sustained until 48 h post oligofructose overload. A reduction in PAF-induced L-selectin shedding was observed at 16 h and 32 h post oligofructose overload. Overall, the results indicated that neutrophil PAF responses were altered in heifers with ruminal acidosis, suggesting a potential dysfunction of the innate immune response.     

Productivity,Respiration, and Light-Response Parameters of World Grassland and Agroecosystems Derived From Flux-Tower Measurements

Grasslands and agroecosystems occupy one-third of the terrestrial area, but their contribution to the global carbon cycle remains uncertain. We used a set of 316 site-years of CO₂ exchange measurements to quantify gross primary productivity, respiration, and light-response parameters of grasslands, shrublands/savanna, wetlands, and cropland ecosystems worldwide. We analyzed data from 72 global flux-tower sites partitioned into gross photosynthesis and ecosystem respiration with the use of the light-response method (Gilmanov, T. G., D. A. Johnson, and N. Z. Saliendra. 2003. Growing season CO₂ fluxes in a sagebrush-steppe ecosystem in Idaho: Bowen ratio/energy balance measurements and modeling. Basic and Applied Ecology 4:167–183) from the RANGEFLUX and WORLDGRASSAGRIFLUX data sets supplemented by 46 sites from the FLUXNET La Thuile data set partitioned with the use of the temperature-response method (Reichstein, M., E. Falge, D. Baldocchi, D. Papale, R. Valentini, M. Aubinet, P. Berbigier, C. Bernhofer, N. Buchmann, M. Falk, T. Gilmanov, A. Granier, T. Grünwald, K. Havránková, D. Janous, A. Knohl, T. Laurela, A. Lohila, D. Loustau, G. Matteucci, T. Meyers, F. Miglietta, J. M. Ourcival, D. Perrin, J. Pumpanen, S. Rambal, E. Rotenberg, M. Sanz, J. Tenhunen, G. Seufert, F. Vaccari, T. Vesala, and D. Yakir. 2005. On the separation of net ecosystem exchange into assimilation and ecosystem respiration: review and improved algorithm. Global Change Biology 11:1424–1439). Maximum values of the quantum yield (α=75 mmol · mol^?1), photosynthetic capacity (A_max=3.4 mg CO₂ · m^?2 · s^?1), gross photosynthesis (P_g,max=116 g CO₂ · m^?2 · d^?1), and ecological light-use efficiency (ε_ecol=59 mmol · mol^?1) of managed grasslands and high-production croplands exceeded those of most forest ecosystems, indicating the potential of nonforest ecosystems for uptake of atmospheric CO₂. Maximum values of gross primary production (8 600 g CO₂ · m^?2 · yr^?1), total ecosystem respiration (7 900 g CO₂ · m^?2 · yr^?1), and net CO₂ exchange (2 400 g CO₂ · m^?2 · yr^?1) were observed for intensively managed grasslands and high-yield crops, and are comparable to or higher than those for forest ecosystems, excluding some tropical forests. On average, 80% of the nonforest sites were apparent sinks for atmospheric CO₂, with mean net uptake of 700 g CO₂ · m^?2 · yr^?1 for intensive grasslands and 933 g CO₂ · m^?2 · d^?1 for croplands. However, part of these apparent sinks is accumulated in crops and forage, which are carbon pools that are harvested, transported, and decomposed off site. Therefore, although agricultural fields may be predominantly sinks for atmospheric CO₂, this does not imply that they are necessarily increasing their carbon stock.     

Plasma atrial natriuretic peptide concentration in warmblood horses with heart valve regurgitations

ObjectivesThis study measured plasma atrial natriuretic peptide (ANP) concentration in horses with heart valve regurgitations (HVR) with and without atrial and ventricular dilatation.BackgroundIn humans and small animals, plasma ANP concentration is increased in heart disease and correlates with the severity of clinical signs and heart enlargement.Animals, materials and methodsTen healthy horses (control) and 36 horses with HVR were evaluated by auscultation, electrocardiography, echocardiography, and determination of plasma ANP.ResultsControl horses demonstrated mean plasma ANP concentration of 21 ± 5.4 pg/mL. Of the 36 horses with HVR, 17 horses possessed normal echocardiographic heart size (group 1), 10 horses had a left atrial dilatation (group 2) and 9 horses had both left atrial and ventricular dilatation (group 3). Mean plasma ANP concentration of groups 1, 2 and 3 was 20.1 ± 5.6 pg/mL, 22.9 ± 11.0 pg/mL and 27.6 ± 17.4 pg/mL, respectively. The plasma ANP concentrations of HVR and control horses were not significantly different. The highest ANP concentrations were observed in horses with atrial and ventricular dilatation. No correlation between left atrial or ventricular size, weight, or sex and the plasma ANP concentration was found.ConclusionsNo significant differences in plasma ANP concentration was observed between groups. Further study, especially in horses with clinical signs of heart failure is needed.