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).     

Persistence of benfluralin in soil and its uptake by carrots

Analytical methods are described for the determination of residues of benfluralin in soil and carrots. All plots treated with 1.3 or 1.9 kg benfluralin/ha produced average carrot yields significantly higher than untreated plots. Degradation of benfluralin in soil was very slow during the winter. As the temperature rose, the rate of benfluralin decline was accelerated; 324 days after incorporation, benfluralin residues were reduced to about 11 and 14% of the applied dose. The average total residues of benfluralin found in whole carrots were 210 ± 18 and 240 ± 47 ng/g following doses of 1.3 and 1.9 kg a.i./ha respectively. The peel of carrots contained 92% of the total residue, and there seemed to be little translocation of the compound through the plant tissues.     

The Neocortex of the Dog

This article describes the structural organization of the neocortex of the dog from purely morphological point of view. Series of 50 urn transverse sections were cut from the frozen right hemiencephala of adult greyhounds, stained by N issl's method, and drawn. His's method was then used to construct life-size lateral, dorsal and medial views of the hemiencephalon from 38 sections located at regular intervals. Once the cytoarchitectonic criteria to be used had been established on the basis of preliminary observations, all the sections of each hemiencephalon were examined microscopically and the cytoarchitectonic types observed in each mapped on the corresponding drawing. These maps were then projected onto lateral, dorsal and medial views of the whole hemiencephalon to produce a classical cytoarchitectonic map of the neocortex made up of 21 types, 7 of them agranular (with no inner granular layer), 2 dysgranular (with a granular layer with diffuse boundans) and the rest granular (with a well-defined layer IV). Various areas of cortex whose characteristics were intermediate between those of adjoinin areas were classified in a residual typus intermedius.     

Gas-liquid chromatographic method for perfluidone residues in rice grains and husks

A gas-liquid chromatographic method using electron-capture detection is described for the determination of perfluidone residues in rice grains and husks. The procedure consists of extraction, purification on an anion-exchange resin, methylation and further clean-up (if necessary) by adsorption on “Florisil”. The recoveries from fortified rice grains and husks were respectively 73.5–98% and 77–104% in the range 0.01–0.50 mg/kg. Sensitivity permits detection of perfluidone at 0.005 mg/kg level.     

Liquid Swine Manure Can Kill Verticillium dahliae Microsclerotia in Soil by Volatile Fatty Acid, Nitrous Acid, and Ammonia Toxicity

ABSTRACT In previous studies, liquid swine manure (LSM) was sometimes shown to reduce Verticillium wilt of potato caused by Verticillium dahliae. We also observed that microsclerotia of this fungus died within 1 day, or between 3 and 6 weeks, after addition of LSM to some acid soils and within 1 week in some alkaline soils. In this study, we demonstrated that a volatile fatty acid (VFA) mixture with an identical concentration of VFAs as that found in an effective LSM reduced germination in an acid soil (pH 5.1) to the same extent as the LSM after 1 day of exposure. Germination was reduced by 45, 75, and 90% in the 10, 20, and 40% ([wt/wt] soil moisture) treatments, respectively, with the latter being equivalent to an application of 80 hl/ha. Addition to this acid soil of 19 LSMs (30% [wt/wt] soil moisture) collected from different producers resulted in complete kill of microsclerotia with 12 manures. Effective manures had a total concentration of nonionized forms of VFAs in soil solution of 2.7 mM or higher. In some acid soils (pH 5.8), addition of LSM (40% [wt/wt] soil moisture) did not kill microsclerotia until 3 to 6 weeks later. Here, a reduction in viability of microsclerotia was attributed to the accumulation of 0.06 mM nitrous acid in the soil solution at 4 weeks. When an LSM was added (40% [wt/wt] soil moisture) to an alkaline soil (pH 7.9) where VFAs are not toxic, microsclerotia germination was reduced by 80% after 1 week. Here the pH increased to 8.9 and the concentration of ammonia reached 30 mM in the soil solution. An ammonium chloride solution having an equivalent concentration of ammonium as the manure was shown to have the same spectrum of toxicity as the manure in assays ranging from pH 7 to 9, both in solutions and above the solutions. At pH 9, the concentration of ammonia reached 18 mM and 100% mortality of microsclerotia occurred. Thus, in acid soils, LSM can kill microsclerotia of V. dahliae by VFA and/or nitrous acid toxicity and in alkaline soils by ammonia toxicity. In order to take advantage of these mechanisms for disease reduction, the manure chemical composition, rate of addition, and soil characteristics need to be determined for each instance of use.     

Preliminary results on detection of maize treated with electrons for seed dressing and according decontamination and infestation

Saatgut und Getreide kann mit niederenergetischen Elektronen (<300 kev) oder=" hochenergetischen=" elektronen=" (1–10 mev)=" wirksam=" behandelt=" werden,=" um=" mikroorganismen=" und=" insekten=" abzutöten.=" in=" dieser=" vorläufigen=" studie=" wurde=" mais=" mit=" niederenergetischen=" (125 kev)=" und=" hochenergetischen=" elektronen=" (10 mev)=" behandelt.=" um=" diese=" elektronenbehandlung=" nachzuweisen,=" wurden=" verschiedene=" verfahren=" eingesetzt:=" photostimulierte=" lumineszenz=" (psl),=" thermolumineszenz=" (tl)=" und=" dna-kometentest.=" für=" diese=" drei=" nachweismethoden=" existieren=" bereits=" europäische=" normen=" und=" sie=" sind=" als=">Allgemeine Codex Methoden zum Nachweis bestrahlter Lebensmittel etabliert. Die Ergebnisse zeigen, dass PSL und TL geeignete Verfahren sind, um sowohl eine Behandlung von Mais mit niederenergetischen als auch mit hochenergetischen Elektronen zu erkennen. Der DNA-Kometentest erwies sich als weniger geeignet: die Behandlung mit niederenergetischen Elektronen konnte—wie erwartet—nicht nachgewiesen werden. Die Behandlung mit hochenergetischen Elektronen konnte bei einer Maissorte erkannt werden, jedoch nicht bei einer anderen Sorte.     

Participation of Rhipicephalus sanguineus (Acari: Ixodidae) in the epidemiology of canine visceral leishmaniasis

The vectorial competence of the tick Rhipicephalus sanguineus is discussed in relation to the epidemiology of canine visceral leishmaniasis, taking into account its strict association with dogs and the low indices of natural infection presented by its known vector, the phlebotomine sand fly Lutzomyia longipalpis. In order to evaluate natural infection by Leishmania chagasi and the infectivity of these parasites in the tick, 39 specimens (6 females, 11 males and 22 nymphs) of R. sanguineus were removed from 21 dogs showing diverse symptoms of zoonotic visceral leishmaniasis (ZVL). Six ticks (15.4%) gave positive results for the genus Leishmania using the PCR technique. To determine the infectivity of the parasites, 36 hamsters were inoculated orally and peritoneally with macerates of ticks removed from nine dogs symptomatic for visceral leishmaniasis. After 6 months the hamsters were sacrificed and necropsied. Serum was removed for IFAT, as well as spleen and liver fragments to make imprint smears and for PCR. Eight (88.9%) of these dogs presented ticks that were infective for 14 hamsters (41.2%), 12 (85.7%) of them infected peritoneally and two (14.3%) orally. PCR revealed 27 smears (40.9%) to be positive, 20 (62.5%) of them infected peritoneally and seven (20.6%) orally. IFAT showed 14 positive animals (41.2%). Based on these findings, we suggest that the vectorial capacity of R. sanguineus for L. chagasi should be evaluated further, opening new perspectives in the epidemiology of ZVL.     

Osteosarcoma with multiple metastases and subcutaneous involvement in a rabbit (Oryctolagus cuniculus)

BACKGROUND: Osteogenic sarcomas are rare in rabbits, but cases involving the mandible, ribs, frontal bones, and tibia have been reported. OBJECTIVE: The purpose of this study was to describe the clinical and pathologic findings in a rabbit with osteosarcoma and multiple metastases with subcutaneous involvement. METHODS: A 1-year-old female Polish rabbit was evaluated for dysorexia, lameness, gait difficulties with the hind limbs, and the presence of 2 subcutaneous nodules, on the left thigh and in the dorsal region. A hard swelling was also present at the base of the tail. Radiographic findings revealed multiple abnormalities in several bones, which were more severe in the sacrococcygeal region and femur. Several areas of calcification involving the thorax and abdomen as well as skeletal muscles, also were noted. The thigh nodule was surgically removed, and cytologic and histologic evaluations were performed. Following the rabbit's death, postmortem radiographs were taken and necropsy was performed. Samples of tissues and organs, involved bone segments, and other subcutaneous nodules were collected and examined by cytology and histology. RESULTS: Both the subcutaneous nodules and the majority of tissues and organs contained neoplastic cells whose appearance was consistent with osteogenic sarcoma. CONCLUSION: Clinical, radiologic, and pathologic data supported a diagnosis of osteosarcoma with multiple metastases and subcutaneous involvement.     

Morphological aspects of in vivo cleavage in Myocastor coypus (coypu)

The objective of the present work was to characterize the in vivo cleavage stage of Myocastor coypus embryos. For this purpose a colpocytological follow-up and controlled mating of 18 females were performed. Specimens from the beginning of the first cleavage to the acquisition of a morula appearance were considered to be in cleavage stage. Embryos in cleavage were collected between days 3 and 6 post-coitus. Of the collected embryos, 80% presented an even number of blastomeres and the remaining 20% an odd number. Embryos from 3 to 7 cells were blastomere associations in a spherical disposition within the zona pellucida. Blastomeres were spherical or ovoid, presenting slight flattening in areas contacting with other blastomeres. Embryos of 8 and 9 cells were as a group of blastomeres slightly elongated, surrounded by a spherical zona pellucida. The percentage of peri-vitelline space occupied by the embryonic mass ranged from 74.1 to 95.8% for all the substages. The cleavage pattern, developed in the oviduct, was of a rotational holoblastic type and asynchronic.     

High-frequency ultrasonography of the skin of clinically normal dogs

OBJECTIVE: To assess the applicability of high-frequency diagnostic ultrasonography for evaluation and accurate measurement of the skin thickness of clinically normal dogs. ANIMALS: 26 healthy dogs (12 sexually intact males, 13 sexually intact females, and 1 spayed female) of various breeds and ages. PROCEDURE: Ultrasonographic examination of the skin and histomorphometric analysis of skin biopsy specimens obtained from the same site were performed. A 13-MHz linear-array transducer was used to obtain a series of ultrasonographic images of the skin in the flank region; images were analyzed and measured by use of imaging software. Cutaneous biopsy specimens were placed in fixative and then stained with H&E and Masson trichrome stains. Histomorphometric analysis was performed by use of an image analyzer. Thickness of the epidermis and dermis of each specimen was evaluated by use of a semiautomatic procedure of quantification. Data obtained from ultrasonographic and histologic measurements were compared by use of the Pearson correlation test. RESULTS: The ultrasonographic pattern of canine skin was consistently characterized by 3 distinct, defined echogenic layers corresponding to the epidermal entry echo, epidermis and dermis, and subcutaneous tissues. A positive correlation was found between ultrasonographic and histologic measurements of skin thickness. CONCLUSIONS AND CLINICAL RELEVANCE: Comparison between ultrasonographic and histologic appearance of the skin revealed that layering of canine skin (ie, epidermis and dermis) and the subcutaneous tissues may be recognized and measured by use of high-frequency ultrasonography. Thus, diagnostic ultrasonography may be a useful tool for the noninvasive evaluation of cutaneous disorders in dogs.