Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Taenia solium oncosphere antigens induce immunity in pigs against experimental cysticercosis

Immunity to Taenia solium infection was investigated using an experimental intramuscular oncosphere infection assay (IMOA) model in pigs. Three naturally infected pigs with cysticercosis were treated with oxfendazole (OFZ), a drug demonstrated to kill cysts in porcine muscle. These animals were then challenged with oncospheres but did not develop any cysts while three uninfected pigs that were similarly challenged, did develop intramuscular cysts. In another study, two groups of three pigs each were immunized with crude T. solium oncosphere and metacestode antigens, respectively, and tested with the IMOA. Immunization with crude oncosphere antigens (OAs) induced 100% protection, while metacestode antigens provided only partial protection. Immunoblots showed that pigs with complete immune protection to oncosphere intramuscular challenge had antibodies to two OAs at 31.3 and 22.5 kDa, respectively. Antibody to these two antigens was absent in pigs immunized with metacestodes or in uninfected control pigs. This study demonstrated the presence of two antigens that are unique to the oncosphere. Although, antibody to these two antigens is consistently present in pigs that are protected from an oncosphere intramuscular challenge their role in preventing infection by T. solium larval cysts is still hypothetical.     

Comparison of echocardiography and gated equilibrium radionuclide ventriculography in the measurements of left ventricular systolic function parameters in healthy dogs

Left ventricular systolic function was assessed in 12 healthy dogs with equilibrium radionuclide ventriculography. The results of the analysis were compared to traditional echocardiographic measurements. Left ventricular internal dimensions and volume were measured at the time of end-systole and end-diastole. Ejection fraction--one of the most informative parameters of cardiac function--was calculated in each animal. Values (e.g. EDD, ESD, EDV, ESV) measured by the scintigraphic method were significantly (Student's t-test, p < 0.05) higher than the data obtained by echocardiography. Ejection fraction (EF) was the only parameter that did not differ significantly when comparing the two imaging techniques. The difference between the results of parallel measurements was in inverse ratio to the size of the heart.     

Major outer membrane proteins of Brucella spp.: past,present and future

The major outer membrane proteins (OMPs) of Brucella spp. were initially identified in the early 1980s and characterised as potential immunogenic and protective antigens. They were classified according to their apparent molecular mass as 36–38 kDa OMPs or group 2 porin proteins and 31–34 and 25–27 kDa OMPs which belong to the group 3 proteins. The genes encoding the group 2 porin proteins were identified in the late 1980s and consist of two genes, omp2a and omp2b, which are closely linked in the Brucella genome, and which share a great degree of identity (>85%). In the 1990s, two genes were identified coding for the group 3 proteins and were named omp25 and omp31. The predicted amino acid sequences of omp25 and omp31 share 34% identity. The recent release of the genome sequence of B. melitensis 16 M has revealed the presence of five additional gene products homologous to Omp25 and Omp31. The use of recombinant protein technology and monoclonal antibodies (MAbs) has shown that the major OMPs appear to be of little relevance as antigens in smooth (S) B. abortus or B. melitensis infections i.e. low or no protective activity in the mouse model of infection and low or no immunogenicity during host infection. However, group 3 proteins, in particular Omp31, appear as immunodominant antigen in the course of rough (R) B. ovis infection in rams and as important protective antigen in the B. ovis mouse model of infection. The major OMP genes display diversity and specific markers have been identified for Brucella species, biovars, and strains, including the recent marine mammal Brucella isolates for which new species names have been proposed. Recently, Omp25 has been shown to be involved in virulence of B. melitensis, B. abortus and B. ovis. Mutants lacking Omp25 are indeed attenuated in animal models of infection, and moreover provide levels of protection similar or better than currently used attenuated vaccine strain B. melitensis Rev.1. Therefore, these mutant strains appear interesting vaccine candidates for the future. The other group 3 proteins identified in the genome merit also further investigation related to the development of new vaccines.     

In vitro and in vivo efficacy of lasalocid for treatment of experimental cryptosporidiosis

In vitro viability of purified Cryptosporidium parvum oocysts, exposed for 30, 60, 90 and 120 min to 0.27 mg/ml lasalocid suspension was evaluated by inclusion or exclusion of two fluorogenic vital dyes and an excystation technique. Continuously, preventive and curative efficacies at different doses (9, 6.75, 5.625 and 4.5 mg/kg body weight) and regimens of lasalocid against cryptosporidial infection were evaluated on an experimental neonatal mice model. In vitro assays demonstrated a decrease in the oocyst viability related to an increase in exposure time for exposure to the lasalocid suspension. The infection was eradicated when the suspension was administered with a dose of ≥6.75 mg/kg body weight. No apparent toxic effects were observed.     

Yield loss in chickpeas in relation to development of fusarium wilt epidemics

ABSTRACT Development of 108 epidemics of Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris were studied on cvs. P-2245 and PV-61 in field microplots artificially infested with races 0 and 5 of F. oxysporum f. sp. ciceris in 1986 to 1989. Disease progression data were fitted to the Richards model using nonlinear regression. The shape parameter was influenced primarily by date of sowing and, to a lesser extent, by chick-pea cultivars and races of F. oxysporum f. sp. ciceris. Fusarium wilt reduced chickpea yield by decreasing both seed yield and seed weight. These effects were related to sowing date, chickpea cultivar, and virulence of the prevalent F. oxysporum f. sp. ciceris race. Regression models were developed to relate chickpea yield to Fusarium wilt disease intensity with the following independent variables: time to initial symptoms (t(is)), time to inflection point (t(ip)) of the disease intensity index (DII) progress curve, final DII (DII(final)), standardized area under DII progress curve (SAUDPC), and the Richards weighted mean absolute rate of disease progression (rho). Irrespective of the chickpea cultivar x pathogen race combination, the absolute and relative seed yields decreased primarily by delayed sowing. The relative seed yield increased with the delay in t(is) and t(ip) and decreased with increasing DII(final), SAUDPC, and rho. A response surface as developed in which seed yield loss decreased in a linear relationship with the delay in t(is) and increased exponentially with the increase of rho.     

Reproductive performance of crossbred and purebred male rabbits

The effect of buck genetic type and crossbreeding parameters on fertility and prolificacy were estimated using two rabbit sire lines and their reciprocal crosses. The relationship between the reproductive performance of inseminated multiparous does and several semen quality traits was also investigated. The semen characteristics evaluated were: pH (pH), mass and individual motility (MM, IM), percentage of viable spermatozoa (Vi), spermatozoa with normal apical ridge (NAR), normal spermatozoa (NSP), spermatozoa with morphological abnormalities of head (HAP), neck-midpiece (NAP), and tail (TAP), spermatozoa with the presence of proximal (PD) and distal (DD) cytoplasmic droplets.

Fertility was analysed as a continuous trait (kindling rate) or as a binary trait (success or failure of kindling). In the first case, the analysis was performed using GLM procedures of SAS v.8 according to a model that included the fixed factors of buck genetic type, number of ejaculates per pool and week of insemination. In the second case, fertility was analysed using GENMOD procedures of SAS v.8 according to a mixed model including the same fixed factors as before plus the physiological status of the does and the permanent random effect of female. Number of kits born alive and number of stillborn were analysed with MIXED procedures of SAS v.8 with the same model used for the analysis of fertility as a binary trait. Estimates of the estimable functions of crossbreeding genetic parameters of the lines were obtained from the solutions of the corresponding models by generalized least squares using GLM, GENMOD and MIXED procedures. Crossbreeding parameters were estimated according to the model of Dickerson. A linear regression was used to determine the relationship between fertility and litter size and the semen characteristics evaluated.

Significant differences in fertility were observed among buck genetic types, which were favourable to type R. Differences between lines in maternal genetic effects were relevant and favourable to type R for fertility. Individual heterosis was important but unfavourable for fertility.

A slight correlation was obtained between all semen quality traits and fertility and prolificacy. Two multiple models were found for fertility, including NAP, IM, NSP, buck genetic type and Vi in one model or NAR in other model. Individual motility had an important positive effect, while NAP had a small negative effect. When MM, TAP and buck genetic type were included in a multiple model for the number of kits born alive, both MM and TAP had significant small effects. Individual motility and DD appeared to be related to number of kits stillborn, but only DD had a significant although negligible effect.     

Heterosis, direct and maternal genetic effects on semen quality traits of rabbits

A complete diallel cross involving two rabbit sire lines (C and R) was carried out to estimate the crossbreeding genetic parameters of seminal traits. 2140 ejaculates from 153 males were analyzed. The traits studied were: presence of gel plugs (G), urine (U), and calcium carbonate deposits (CC), number of useful ejaculates (UE), pH, volume (V), mass and individual motility (Mm, Mi), useful Mi (UM), concentration (Cn), number of spermatozoa per ejaculate (TSE), percentage of viable spermatozoa (Vi), spermatozoa with normal apical ridge (NAR), normal spermatozoa (Nr), spermatozoa with morphological abnormalities of head (H), neck-midpiece (Nm), and tail (T), presence of proximal and distal cytoplasmic droplet (Dp, Dd).

Estimates of heterosis, direct and maternal genetic effects were obtained from the solutions of the mixed model. There were major differences in direct genetic effects between lines, which were favourable to line C for Cn and TSE, and unfavourable for CC, Nm and Dp. Smaller differences were also observed in Vi and NAR favourable to line R. Differences between lines with respect to the maternal genetic effects were relevant and favourable to line C for V and to line R for U, UM, Cn, TSE, Nm, Mi, and Mm. Individual heterosis was high for Dp and Dd.