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).     

Hemangiosarcoma of the third eyelid in a cat

A case of hemangiosarcoma of the third eyelid in a 15-year-old Domestic Short-haired cat is reported. A small red mass protruding from the anterior surface of the third eyelid in the left eye was present. Only mild bilateral conjunctivitis and nuclear sclerosis were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery and cryotherapy were performed. On histopathologic examination the mass was located just beneath the conjunctival epithelium, suggesting that the conjunctiva was the tissue of origin. The tumor showed high cellularity and was composed of pleomorphic spindle and polygonal cells arranged in interlacing bundles and solid sheets. The presence of slit-like spaces between tumor cells containing erythrocytes demonstrated the vascular differentiation. Mitotic activity was moderate, but atypical mitoses were detected. The surgical area healed uneventfully and after 7 months there was no recurrence. To the authors' knowledge, this is the first report of conjunctival hemangiosarcoma of the third eyelid in a cat.     

Restriction endonuclease and monoclonal antibody analysis of Brazilian isolates of bovine herpesviruses types 1 and 5

Twelve Brazilian isolates and three reference strains of bovine herpesviruses (BHVs) were subjected to restriction endonuclease analysis (REA) and monoclonal antibody (MAb) analysis. Viral DNA was cleaved with BamHI, BstEII, EcoRI, HindIII and PstI. The monoclonal antibody panel allowed the differentiation between types 1 and 5 viruses, while REA with BstEII and HindIII showed the distinction between BHV-1 and -5 subtypes. Typical 1.1 and 1.2a patterns were observed with two isolates from respiratory disease. An isolate from semen of a clinically healthy bull displayed 1.2b profile, whereas another displayed a clear 5a pattern, which was never reported before. Seven out of nine Brazilian type 5 (BHV-5) isolates displayed REA patterns similar to the Australian BHV-5 strain N569 (BHV-5a), and differing from the Argentinean A663 strain (BHV-5b) virus. Another two BHV-5 isolates, which displayed an unusual MAb pattern of reactivity, showed a BstEII profile different from both reference strains of BHV-5. These two viruses were considered BHV-5 "non-a/non-b" subtype.     

Occurrence of various immunoglobulin isotopes in horses with equine recurrent uveitis (ERU)

We investigated 30 healthy eyes and 41 eyes with ERU from 57 horses. The total immunoglobulin titers and titers of IgGa, IgGb, IgM were measured in aqueous humour, vitreous and serum using different ELISA techniques. Every sample investigated contained detectable amounts of immunoglobulins. Compared to control eyes significantly increased titers were found in the aqueous humour and vitreous of the ERU eyes for all immunoglobulin isotypes studied (p < or = 0.01). While IgM was detected in only 2 out of thirty aqueous humour and in none of the thirty vitreous samples of healthy eyes, 79.6% of samples of ERU eyes revealed considerable IgM titers. Changes of the IgGa/IgGb ratio in the eye as compared to that in the autologous serum was more frequent in affected than in healthy eyes. In contrast to the intraocular immunoglobulins there were no significant differences in immunoglobulin serum titers in healthy horses and those affected with ERU (p > 0.05). In conclusion, the results argue for a physiological appearance of immunoglobulins in the healthy eye. The increased titers of immunoglobulins in eyes stricken with ERU might be signs either of a local ocular production of antibodies and/or an increased permeability of intraocular barriers.     

Effect of the previously gravid uterine horn and postpartum interval on follicular diameter and conception rate in beef cows treated with estradiol benzoate and progesterone

An experiment was performed to evaluate the effect of the side of ovulation with respect to the previously gravid uterine horn on fertility of cows inseminated at one of two periods postpartum. All cows were treated with an intravaginal progesterone insert for 7 d and received estradiol benzoate (2 mg, i.m.) at the time of device insertion, prostaglandin F2alpha (25 mg, i.m.) at the time of device removal, and estradiol benzoate (1 mg, i.m.) 30 h after device removal. All cows were inseminated 28 to 30 h after the second treatment with estradiol benzoate, regardless of observed estrus. Cows treated in Period 1 received inserts at 16 to 20 d postpartum and were inseminated at 25 to 29 d postpartum. Cows treated in Period 2 received inserts at 26 to 30 d postpartum and were inseminated at 35 to 39 d postpartum. Diameter of the largest follicle at insert removal was greater in cows treated in Period 2 (10.1 +/- 0.3; mm +/- SEM) than in cows treated in Period 1 (9.1 +/- 0.3; P < .05). Diameter did not differ with the side of ovulation in respect to the previously gravid uterine horn. Diameter was greater in cows 5 to 9 (10.3 +/- 0.3) than in cows 3 to 4 (9.0 +/- 0.3) or 10 to 13 (9.4 +/- 0.6) yr of age (P < .01). The proportion of cows that ovulated from the ovary contralateral to the previously gravid uterine horn was greater (P < .05) than of those that ovulated from the ipsilateral ovary, and the incidence of ovulation was reduced in cows 3 to 4 yr of age (P < .01). Conception rate tended to be greater for ovulation from the ipsilateral compared with the contralateral ovary, relative to the previously gravid uterine horn (P < .10) and for ovulation from the right than the left ovary (P < .06). Conception rate was less if cows ovulated a follicle that was < 9 mm than a follicle > or = 9 mm in diameter at insert removal (P < .01) and was greater in cows inseminated in June than in April or May (P < .05). In conclusion, in cows in which estrus was synchronized at 25 to 39 d postpartum, ovulation from either the ovary ipsilateral to the previously gravid uterine horn, or the right ovary, tended to increase fertility.     

Expression and characterisation of equine interleukin 2 and interleukin 4

In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His(6)-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.     

Canine echinococcosis: an alternative for surveillance epidemiology

The essential activities for programmes of cystic echinococcosis control are the census of all dogs from the program and identification of parasitised animals. Currently, in South America evaluations and epidemiological surveillance are based on the administration of arecoline hydrobromide. This method has the disadvantage of increasing environmental pollution and risk for operators and owners of treated dogs. A genus-specific ELISA capture method has been employed for recently issued faeces and the confirmation of positive examination was performed by dog autopsies. Our work presents an alternative method based on collection of dry field-dispersed faeces, followed by serological diagnosis by Copro-ELISA and confirmation by Copro-Western blot. If Copro-ELISA were used to define positive samples of dry faeces, the Copro-Western blot assay would provide 70% sensitivity and 100% specificity. Global efficiency of the system using dry faeces would reach 76%, allowing epidemiological surveillance to be oriented to analysis of surface units instead of dog as measurement unit.     

An epizootiological survey of necropsy cases (1993-1997) at University of the Philippines

An epizootiological survey of necropsied cases (1993-1997) at University of the Philippines was performed. A total of 368 cases included 238 avian and 111 porcine cases. Amongst avian cases, the major cause of death was infectious diseases in 212 (89%) cases including 97 (41%) bacterial, 36 (15%) viral, and 21(9%) parasitic diseases. The majority of the avian bacterial diseases presented as septicemia (73 cases) and the viral diseases as Newcastle disease (17 cases). In porcine cases, the major cause of death was also infectious diseases, in 100 (90%) cases including 52 bacterial and 29 viral diseases. Porcine bacterial diseases were classified into 36 septicemia, 4 hemophillosis and 4 colibacillosis. Amongst the porcine viral diseases, most cases were diagnosed as Hog cholera (22 cases).     

Neue Ansätze zur Optimierung der Kraut- und Knollenfäulebekämpfung im ökologischen Kartoffelanbau

Potato late blight is still an unsolved problem in organic farming. Up to now the disease can only be controlled by copper fungicides. Our project is aiming to reduce the application of copper-containing pesticides by introduction of a blight forecasting system based on meteorological parameters. Primary stem infections should be reduced by seed treatment with copper fungicides thus to postpone the beginning of the blight epidemic as well as the start of spraying. To control secondary infections on the foliage, fungicide strategies should be elaborated to achieve best efficacy with reduced amounts of pesticides. Therefore copper amounts and spraying intervals should be adjusted to the infection pressure. Based on the biological and epidemiological conditions for primary and secondary infections with special emphasis on soil moisture, the potato blight forecast system ÖKO-SIMPHYT should be developed.     

The socioeconomics and management of Santiago de Chile''s public urban forests

Santiago, Chile's semi-arid climate and urbanized environment poses a severe limitation for the establishment and maintenance of urban forests. Municipalities, or comunas, are the main stakeholders in the management of Santiago's public urban forests. A tenable hypothesis would be that as the socioeconomic level of a comuna increases, the better the condition of a comuna's urban forest. Unfortunately, there is little comprehensive information on management, public expenditure, and structure of Santiago's public and private urban forests. To examine this hypothesis, Santiago was divided into socioeconomic strata, then using air photo interpretation and stratified field sampling, urban forest structures were quantified by socioeconomic strata. In addition, interview surveys were used to determine municipal urban forest management and expenditures for different public urban forests based on socioeconomic strata. Urban forests in the high socioeconomic strata had fewer public trees, greater tree cover, tree and leaf area density, and leaf area index than lower socioeconomic strata. The percentage of total municipal budget allocated to public urban forest management was consistent among strata, but the total public urban forest budgets were greater in the high socioeconomic strata. Public urban forest structure is related to the socioeconomic strata of Santiago's different comunas.