Pathological analysis of batch safety testing of veterinary vaccines using small laboratory animals

Batch safety tests (BSTs) of veterinary vaccines are conducted using small laboratory animals to assure the safety of vaccines according to several criteria, including clinical signs and change in body weight. Although the latter is used as an evaluation index in BSTs, there have been no reports on the internal changes that affect body weight during the test period. Therefore, we analyzed BST via pathological examination of the tested animals. Here, BSTs were performed for 176 batches using mice and 126 batches using of guinea pigs. Most of the gross findings could be classified into four lesion types (nodules, adhesions, ascites, no apparent signs), with only one vaccine inducing lesions that could not be classified into any of these four types. Histopathological examination revealed that the reactions caused by BST were pyogenic and/or granulomatous inflammation. Nodular or adhesive lesions comprised more severe pyogenic granulomatous inflammation than ascites or cases with no apparent gross lesions. These nodular or adhesive lesions were more frequently induced by vaccines that contained an adjuvant than by vaccines that did not contain an adjuvant. The cases with “exceptional” gross findings histologically presented severe necrosis of the hematopoietic system. Additional testing showed that these “exceptional” lesions were induced when a specific type of light liquid paraffin was injected along with other vaccine additives. Our results show that body weight loss and/or lesions during BST were induced by proinflammatory properties of the tested vaccines and that BST is a sensitive method for detecting unexpected effects of vaccine components.     

Expression of P-glycoprotein and breast cancer resistance protein in three cases of canine lymphoma showing drug resistance

In canine lymphoma, drug resistance is the major factor hindering treatment. In this study, we performed immunohistochemical examination of P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), which are considered as transporters related to multidrug resistance in three recurrent canine lymphomas. All cases were negative for both transporters before anticancer drug administration, but became positive after this administration. The expression was confirmed in capillary endothelial cells, such as in brain capillaries acting as the blood-brain barrier (BBB). It is suggested that both transporters expressed on capillary endothelial cells in lymphoma tissue may inhibit the spread of anticancer drugs into tumor tissues from blood, the same as the BBB. Therefore, capillary endothelial cells could act as a blood-tumor barrier, which might be involved in drug resistance in canine lymphoma.     

Canine fetal echocardiography: correlations for the analysis of cardiac dimensions

The aim of this study was to develop regression models for correlation of canine fetal heart development with body size to characterize normal development or suggest cardiac anomalies. Twenty clinically healthy pregnant bitches, either brachycephalic and non-brachycephalic, were examined ultrasonographically. Transabdominal fetal echocardiography was conducted every 4 days from the beginning of cardiac chambers differentiation until parturition. Ten cardiac parameters were measured: length, width and diameter of the heart; heart area; left and right ventricular dimensions; left and right atrial dimensions; and aortic and pulmonary artery diameter. Femoral length, biparietal diameter and abdominal cross-sectional area were also recorded. Regression equations were developed for each parameter of fetal body size, and linear and logarithmic models were compared. The model with the highest correlation coefficient was chosen to produce equations to calculate relative dimensions based on the correlations. Only the left-ventricular chamber differed between the two racial groups. Biparietal diameter was the independent parameter that produced the highest correlation coefficient for the most fetal cardiac dimensions, although good correlations were also observed using femoral length and abdominal cross-sectional area. Heart width and heart diameter were used as surrogates of cardiac development, as these measurements showed the best statistical correlation. Quantitative evaluation of fetal cardiac structures can be used to monitor normal and abnormal cardiac development.     

Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

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.

     

Involvement of a host erythrocyte sialic acid content in Babesia bovis infection

Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant alpha2-3-linked SA residues but not alpha2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of alpha2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining with the alpha2-3-linked SA-specific lectin: a selective invasion of B. bovis was found only in the intact RBCs bearing the SAs, but not in the desialylated RBCs. Furthermore, a significant reduction of the parasitized RBCs was also observed in the culture supplemented with exogenous 3'-sialyllactose containing the alpha2-3-linked SAs. However, the complete inhibition of parasite proliferation was not achieved in the culture. These findings indicate that while the alpha2-3-linked SA-dependent pathway is needed for highly efficient invasion of host RBCs by B. bovis, there might also be other potential alternative pathways.     

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.     

Crop response of drought-tolerant and conventional maize hybrids in a semiarid environment

In the High Plains, corn (Zea mays L.) is an important commodity for livestock feed. However, limited water resources and drought conditions continue to hinder corn production. Drought-tolerant (DT) corn hybrids could help maintain high yields under water-limited conditions, though consistent response of such hybrids is unverified. In this two-year study, the effects of three irrigation treatments were investigated for a DT and conventional maize hybrid, Pioneer AQUAMax P0876HR and Pioneer 33Y75, respectively. In 2013, the drier of the 2 years, irrigation amounts and crop water use (ET_c) were greater for the conventional hybrid, but grain water use efficiency (WUE) and harvest index were significantly greater for the DT hybrid. In 2014, grain yields and WUE were not significantly different between hybrids. However, irrigation amounts, ET_c and biomass yields were greater for the conventional hybrid. Results from both years indicate that the DT hybrid required less water to maximize grain yield as compared to the conventional hybrid. Producing relatively high yields with reduced amounts of water may provide a means for producers to continue corn production in a semiarid environment with declining water supplies.