Identification of transgenes from wood of genetically transformed poplar trees

We have extracted total DNA from different fractions of fresh wood as well as from cold-stored and air-dried wood harvested from transgenic aspen grown in the field. The highest amounts of DNA were obtained from bark/cambium tissue; the DNA quality, however, was poor. Best results in PCR and Southern blot analyses were obtained from DNA extracted from early wood. Using appropriate primer pairs, amplification products were obtained from both the foreign gene (transgene) and aspen genomic sequences. In Southern blot analyses transgene-specific hybridisation signals were obtained. This is the first report on the detection of foreign genes in wood sampled from genetically modified trees.     

Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings

Clinical signs, humoral and cellular immune responses, and microscopic and gross tissue alterations resulting from acute experimental Ehrlichia canis infection in dogs were studied. Four dogs were inoculated with E. canis and four were used as uninfected controls. After a 10-14-day incubation period, infected dogs developed pyrexia up to 41 degrees C for 6-8 days. Antibody titers to E. canis antigen were demonstrable in all inoculated dogs at 30 days post-infection. Necropsy of infected animals revealed pale mucous membranes, generalized lymphadenopathy, splenomegaly, edema and ascites. Microcopically, the main lesions were: lymphoreticular hyperplasia in cortical areas of lymph nodes and spleenic white pulp, periportal accumulation of mononuclear cells and centrolobular fatty degeneration of the liver. Kidneys presented with glomerulonephritis characterized by interstitial mononuclear infiltration. Immunophenotyping of lymphocytes from lymph nodes and spleen sections displayed alterations in IgG, IgM, CD3+ and CD8+ cells population in infected dogs.     

Seroprevalence of Toxoplasma gondii antibodies from wild canids from Brazil

The prevalence of anti-Toxoplasma gondii antibodies was evaluated by the indirect immunofluorescent-antibody test in serum of 57 wild canids from three different species: Lycalopex gymnocercus, Cerdocyon thous and Dusicyon vetulus from the northeast, southeast and southern regions of Brazil. The prevalence was 35.1%, with 20 of the 57 canids demonstrating antibodies anti-T. gondii at dilutions of 1:16 in 2, 1:32 in 4, 1:64 in 2, 1:128 in 2, 1:256 in 6, 1:512 in 2 and 1:2048 in 2 animals. None of the D. vetulus were positive. Among the L. gymnocercus 11 (91.7%) of the 12 samples were positive and among C. thous 9 (60%) of the 15 had antibodies anti-T. gondii.     

Comparison of the sensitivity of in vitro and in vivo tests for detection of the presence of a bovine viral diarrhoea virus type 1 strain

Veterinary vaccines are usually tested for the absence of contaminants. However, the quality control does not always imply that vaccines are not contaminated as, for example, illustrated by the bovine herpes virus 1 (BHV1) vaccine used in The Netherlands in 1999 that contained a small amount of bovine viral diarrhoea virus (BVDV1). Thousands of cows were vaccinated with BHV1 vaccine batches, and the question arose as to whether these small amounts of BVDV1, most likely not detected with in vitro tests, could have infected cattle. More in general, the question was whether the outcome of the in vitro tests, i.e. the in vitro infectivity, was indicative for the infectivity for cattle, i.e. the in vivo infectivity. We therefore carried out in vitro experiments to determine the sensitivity of a BVDV1 isolation assay. In addition, we performed two animal experiments, in which we estimated the lowest dose needed to infect calves with BVDV1. We extrapolated the experimental in vitro and in vivo results from a tissue culture infectious dose (TCID50) to a cattle infectious dose (CID50). We observed a partial response in the calves inoculated with this dose: four out of six calves turned out to be infected. In the tissue culture test, all 20 samples tested negative. The response in vivo, however, was not significantly higher than the in vitro response, which implies that no difference in susceptibility was observed between the animal test and the tissue culture test. Based on the results in our experiments, some cattle may have been infected with BVDV1 after the application of the contaminated BHV1 vaccine during the vaccination campaign. The question remains that how many cattle received contaminated vaccine, and became infected with BVDV1.     

High urinary corticoid/creatinine ratios in cats with hyperthyroidism

Measurement of the urinary corticoid : creatinine (C : C) ratio provides an assessment of cortisol secretion over a period of time. Therefore, this test is a very sensitive measure of adrenocortical function. The stress of the diagnostic procedure and nonadrenal disease may increase the urinary C : C ratio. In addition, diseases such as hyperthyroidism may influence the metabolic clearance of cortisol. To evaluate the effect of thyroid hormone excess, urinary C : C ratios were measured in 32 cats with hyperthyroidism and 45 healthy household cats. In 7 cats, urinary C : C ratios were measured both before and after treatment for hyperthyroidism. With data from the healthy cats, the reference range for the urinary C : C ratio was determined to be 8.0 to 42.0 X 10(-6). The urinary C : C ratios in the cats with hyperthyroidism (median, 37.5 x 10(-6); range, 5.9-169.5 x 10(-6)) were significantly (P = .001) higher than those in the healthy cats (median, 16 x 10(-6); range, 4.8-52.5 x 10(-6)). In 15 cats with hyperthyroidism, the urinary C : C ratios exceeded the upper limit of the reference range. Treatment for hyperthyroidism led to a marked decrease in urinary C : C ratios. The results of this study demonstrate that the urinary C : C ratio may be abnormally high in cats with hyperthyroidism, probably because of increased metabolic clearance of cortisol and activation of the pituitary-adrenocortical axis by disease. Although the clinical features of hyperthyroidism and hyperadrenocorticism in cats are different, hyperthyroidism should be ruled out when cats are suspected of hyperadrenocorticism on the basis of abnormally high urinary C : C ratios.     

Assessment of the effects of age and joint disease on hydroxyproline and glycosaminoglycan concentrations in synovial fluid from the metacarpophalangeal joint of horses

OBJECTIVE: To assess the effects of age and joint disease on hydroxyproline and glycosaminoglycan (GAG) concentrations in synovial fluid from the metacarpophalangeal joint of horses and evaluate the association of those concentrations with severity of osteoarthritis and general matrix metalloproteinase (MMP) activity. SAMPLE POPULATION: Synovial fluid was collected from the metacarpophalangeal joints of foals at birth (n = 10), 5-month-old foals (10), 11-month-old foals (5), and adult horses (73). PROCEDURE: Hydroxyproline and GAG concentrations were determined in synovial fluid samples. The severity of osteoarthritis in adult joints was quantified by use of a cartilage degeneration index (CDI) and assessment of general MMP-activity via a fluorogenic assay. RESULTS: Hydroxyproline and GAG concentrations in synovial fluid were highest in neonates and decreased with age. Concentrations reached a plateau in adults by 4 years and remained constant in healthy joints. In synovial fluid from osteoarthritic joints, hydroxyproline and GAG concentrations were not increased, compared with unaffected joints, but hydroxyproline were significantly correlated with the CDI and general MMP activity. There was no significant correlation between GAG concentration and CDI value or MMP activity. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in hydroxyproline concentration in synovial fluid appeared to indicate damage to collagen of the articular cartilage. In joints with osteoarthritis, the lack of high GAG concentration in synovial fluid and the absence of a significant correlation between GAG concentration and CDI values or MMP activity may severely limit the usefulness of this marker for monitoring equine joint disease.     

Effect of diet composition and feeding pattern on the prececal digestibility of starches from diverse botanical origins measured with the mobile nylon bag technique in horses

This trial was conducted to determine the extent of prececal starch digestibility depending on the botanical origin of starch and on diet characteristics (i.e., composition and feeding pattern). The prececal disappearance of six substrates (oats, barley, corn, horse bean, potato, and wheat) was measured in four cannulated horses fed (as-fed basis) 11.8 g/kg BW of a high-fiber (HF) or high-starch (HS) pelleted feed and 10.0 g/kg BW of meadow hay using the mobile bag technique (MBT). The daily feeding pattern was either three meals (two meals of pellets and one meal of hay) or five meals (three meals of pellets and two meals of hay). The experimental procedure was a 2 x 2 factorial arrangement tested in a Latin square design. After 2 wk of adaptation to the diet, collections were made on 5 d. Thirty nylon bags, composed of five bags of each substrate, were intubated to each horse during the ingestion of the morning meal. Bags were collected in the cecum, using a magnet, at 9 h postintubation. In spite of strong interindividual differences, approximately 80% of the intubated bags were collected. On average, the mean retention time of the bags was 6.2 h (+/-0.17). Regardless of the feeding pattern, the transit of the bags was faster when the fiber content of the diet was higher (P = 0.003). Likewise, regardless of the meal composition, transit was also faster when the ration was split into five daily meals (P = 0.001). The DM disappearance, corrected with particulate losses (DMD(c)), differed depending on the substrate tested (33.5, 57.1, 63.8, 67.7, 78.6, and 86.2% for potato, horse bean, oats, barley, corn, and wheat, respectively; P = 0.001). The DMD(c) of corn, barley, and potato was higher when HS was fed (P = 0.020); regardless of the substrate, DMD(c) was higher with five daily meals (P = 0.001). The starch disappearance (StarchD(c)) was different depending on the substrate (P = 0.001; 36.1, 71.2, 86.6, 89.2, 99.0, and 99.7% for potato, horse bean, barley, corn, wheat, and oats, respectively). Whatever the substrate, StarchD(c) was higher when HS was fed (P = 0.007), but it was not affected by the feeding pattern of the diet. Although passage rate was modified and feed intake was different, the botanical origin of starch was the main factor that affected prececal starch disappearance in horses.     

Helminths in a giraffe (Giraffa camelopardalis xgiraffa) from a zoo in Spain

A pregnant female Cape giraffe (Giraffa camelopardalis giraffa) died from an unknown cause in the Aitana Zoo, Alicante, Spain. Neither clinical signs nor macroscopic lesions were observed at necropsy. The alimentary tract was removed and examined for parasites. A total of 2 724 nematodes were found, including Camelostrongylus mentulatus, Trichostrongylus axei, Ostertagia ostertagi, Teladorsagia circumcincta, Teladorsagia trifurcata, Marshallagia marshalli, Trichostrongylus vitrinus, Trichostrongylus colubriformis, Spiculopteragia asymmetrica and Trichuris giraffae. Only T. giraffae and C. mentulatus have been previously reported from giraffes. The other nematodes are common in mouflons, fallow and red deer, which can usually be found in the same paddock as the giraffes in Aitana Zoo. Although its occurrence is unusual in this host, C. mentulatus was the most abundant nematode in our giraffe. This parasite has been related to disease, and even death, in several wild ruminants.