Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

Evaluation of truncated LipL32 expressed by Escherichia coli and Pichia pastoris for serodiagnosis of Leptospira infection in rodents

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.     

Assessment of rabbit spleen size using ultrasonography

Assessment of spleen size using the ultrasonography has become a standard practice in human. However, the assessment is not established method in experimental animals. To establish the index to assess the spleen size using ultrasonography, we measured the cross-section image of rabbit spleen during endotoxin shock. The image of the cross-section was appeared as triangle, and the height of the triangular image was defined as the spleen index. This spleen index showed strong correlation with the spleen weight. In conclusion, this method is suitable for observation of changes in rabbit spleen size and may reduce the number of rabbit in the longitudinal studies.     

Cyclosporine A inhibits the mRNA expressions of IL-2, IL-4 and IFN-gamma, but not TNF-alpha, in canine mononuclear cells

The effects of the calcineurin inhibitors cyclosporine A (CsA) and FK506 on the mRNA expressions of various cytokines were evaluated in dogs to determine whether the effects of CsA and FK506 in dogs were similar to those in humans. The mRNA expression levels of the cytokines IL-2, IL-4, IFN-gamma and TNF-alpha were measured in PHA-stimulated canine PBMC using real-time RT-PCR after incubation with CsA or FK506 for 5 hr. Both reagents inhibited IL-2, IL-4 and IFN-gamma mRNA expressions in a dose-dependent manner. However, CsA hardly inhibited the mRNA expression of TNF-alpha. These findings are important for assessing the indications of CsA treatment in dogs.     

A case of hyperplastic dermatosis of the West Highland White Terrier controlled by recombinant canine interferon-gamma therapy

A 3.5-year-old, male West Highland White Terrier was diagnosed as having hyperplastic dermatosis by clinical and histopathological findings. Controlling of Malassezia overgrowth by antifungal drugs provided a temporal improvement of the skin lesions, but the disease was deteriorated within the next 2 months despite the negative demonstration of the yeasts. Induction of recombinant canine interferon-gamma (rCaIFN-gamma) therapy resulted in almost complete cure of the skin lesions within 2 months after the initiation of the therapy. No adverse effects were detected during the therapy. Our results suggested that the rCaIFN-gamma therapy is potential to be a novel therapeutic option for controlling the breed-specific hyperplastic skin disease.     

Using midday stem water potential for scheduling deficit irrigation in mid–late maturing peach trees under Mediterranean conditions

Irrigation techniques that reduce water applications are increasingly applied in areas with scarce water resources. In this study, the effect of two regulated deficit irrigation (RDI) strategies on peach [Prunus persica (L.) Batsch cv. “Catherine”] performance was studied over three growing seasons. The experimental site was located in Murcia (SE Spain), a Mediterranean region. Two RDI strategies (restricting water applications at stage II of fruit development and postharvest) based on stem water potential (Ψ_s) thresholds (?1.5 and ?1.8 MPa during fruit growth and ?1.5 and ?2.0 MPa during postharvest) were compared to a fully irrigated control. Soil water content (θ_v), Ψ_s, gas exchange parameters, vegetative growth, crop load, yield and fruit quality were determined. RDI treatments showed significantly lower values of θ_v and Ψ_s than control trees when irrigation water was restricted, causing reductions in stomatal conductance and photosynthesis rates. Vegetative growth was reduced by RDI, as lower shoot lengths and pruning weights were observed under those treatments when compared to control. However, fruit size and yield were unaffected, and fruit quality was slightly improved by RDI. Water savings from 43 to 65 % were achieved depending on the year and the RDI strategy, and no negative carryover effect was detected during the study period. In conclusion, RDI strategies using Ψ_s thresholds for scheduling irrigation in mid–late maturing peach trees under Mediterranean conditions are viable options to save water without compromising yield and even improving fruit quality.     

Measured and estimated water use and crop coefficients of grapevines trained to overhead trellis systems in California’s San Joaquin Valley

The use of overhead trellis systems for the production of dry-on-vine (DOV) raisins and table grapes in California is expanding. Studies were conducted from 2006 to 2009 using Thompson Seedless grapevines grown in a weighing lysimeter trained to an overhead arbor trellis and farmed as DOV raisins for the first two years and for use as table grapes thereafter. Maximum canopy coverage for the two lysimeter vines across years was in excess of 80 %. Seasonal (15 March–31 October) evapotranspiration for the lysimeter vines (ET_Lys) was 952 mm in 2007 (farmed as DOV raisins) and 943 and 952 mm (when farmed as table grapes). The maximum crop coefficient (K _cLys) across all 4 years ranged from 1.3 to 1.4. These maximum values were similar to those estimated using the relationship where K _c is a function of the amount of shaded area measured beneath the canopy at solar noon (K _c = 0.017 × percent shaded area). Covering the lysimeter’s soil surface with plastic (and then removing it) numerous times during the 2009 growing season (1 June–14 September) reduced ET_Lys from an average of 6.4 to 5.6 mm day^?1 and the K _c from 1.07 to 0.93. A seasonal basal K _c (K _cb) was calculated for grapevines using an overhead trellis system with a 13 % reduction in the K _cLys across the growing season.     

Modeling soil water dynamics in a drip-irrigated intercropping field under plastic mulch

Intercropping, drip irrigation, and the use of plastic mulch are important management practices, which can, when utilized simultaneously, increase crop production and save irrigation water. Investigating soil water dynamics in the root zone of the intercropping field under such conditions is essential in order to understand the combined effects of these practices and to promote their wider use. However, not much work has been done to investigate soil water dynamics in the root zone of drip-irrigated, strip intercropping fields under plastic mulch. Three field experiments with different irrigation treatments (high T1, moderate T2, and low T3) were conducted to evaluate soil water contents (SWC) at different locations, for different irrigation treatments, and with respect to dripper lines and plants (corn and tomatoes). Experimental data were then used to calibrate the HYDRUS (2D/3D) model. Comparison between experimental data and model simulations showed that HYDRUS (2D/3D) described different irrigation events and SWC in the root zone well, with average relative errors of 10.8, 9.5, and 11.6 % for irrigation treatments T1, T2, and T3, respectively, and with corresponding root mean square errors of 0.043, 0.035, and 0.040 cm³ cm^?3, respectively. The results showed that the SWC in the shallow root zone (0–40 cm) was lower under non-mulched locations than under mulched locations, irrespective of the irrigation treatment, while no significant differences in the SWC were observed in the deeper root zone (40–100 cm). The SWC in the shallow root zone was significantly higher for the high irrigation treatment (T1) than for the low irrigation treatment, while, again, no differences were observed in the deeper root zone. Simulations of two-dimensional SWC distributions revealed that the low irrigation treatment (T3) produced serious severe water stress (with SWCs near the wilting point) in the 30–40 cm part of the root zone, and that using separate drip emitter lines for each crop is well suited for producing the optimal soil water distribution pattern in the root zone of the intercropping field. The results of this study can be very useful in designing an optimal irrigation plan for intercropped fields.     

Atmospheric Mercury Fluxes in a Southern Boreal Forest and Wetland

Total gaseous mercury (TGM) fluxes from the forest floor and a boreal wetland were measured by a flux chamber technique coupled with an automatic mercury vapour analyser. The fluxes were measured at three sampling sites in southern Finland, 61°14′ N, 25°04′ E in summer 2007, with additionally in situ TGM concentrations in the air at one of the sites and mercury bulk deposition at another. Most of the flux data were collected during the daytime. At one of the sites, diurnal flux behaviour was studied, and a clear cycle with an afternoon maximum and a night minimum was observed. The highest emissions (up to 3.5 ng m⁻² h⁻¹) were observed at the forest floor site having a moss and grass cover. At the wetland and litter-rich forest floor sites, the emissions were below 1 ng m⁻² h⁻¹ and sometimes negative (down to −1.0 ng m⁻² h⁻¹), indicating mercury uptake. The measured average fluxes in August were 0.9 ± 1.1 and 0.2 ± 0.3 ng m⁻² h⁻¹ for the forest floor sites and wetland sites, respectively. The flux data were compared with the mercury bulk deposition, which proved to be of the same magnitude, but opposite in sign. At the mossy forest floor site, the extrapolated TGM emissions were 130% of the Hg deposition in August 2007. Comparison with other studies showed that the fluxes in background areas are relatively uniform, regardless of measurement site location and method used. Airborne TGM remained at the background level during the study, with an average value of 1.3 ± 0.2 ng m⁻³; it frequently showed a diurnal cycle pattern.