Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva.

Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purfied swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-Il) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 +/- 0.017 mg/mg total protein.     

Comparison of three in vitro techniques to estimate benzimidazole resistance in Haemonchus contortus of sheep

Three in vitro assays to detect benzimidazole resistance, namely, the egg-hatch assay, tubulin-binding assay, and a larval-development assay, were evaluated by estimating the level of benzimidazole resistance in three field isolates of Haemonchus contortus compared with a susceptible reference strain. Comparisons were also made with estimates of benzimidazole resistance of the three field strains obtained from an in vivo controlled anthelmintic efficacy test. All three in vitro tests showed similar, consistent results which also suggested greater sensitivity than the in vivo assay. These results indicate that selection of an in vitro technique to determine benzimidazole resistance should therefore be based on considerations other than precision, such as technical expertise, availability of equipment, cost and speed in which diagnosis is required.     

The Effect of Nitrogen and Phosphorus Fertilization on the Nitrogen Absorption by Sunflowers

Little is known about the effect of fertilization on the N uptake of sunflowers. A 4² factorial trial with 0, 60, 120 and 180 kg N ha⁻¹ and 0, 15, 30 and 45 kg P ha⁻¹ was conducted over three years. The N content and concentration of leaves, stems and capitula were determined at three growth stages. High N levels increased the N content and concentration of all plant parts at all growth stages sharply. High P levels increased the N content of all plant components through better growth. P has an inconsistent effect on N concentration but tended to decrease it. After flowering the crop assimilated 20 to 25 % of the total N. This implies that N applied can still be applied and utilized by the crop at a late stage. This should be substantiated by further research.     

On the genetic basis of equine allergic diseases: II. Insect bite dermal hypersensitivity.

The horses studied were of the Swiss Warmblood breed and most were ELA-typed to assess a possible association of dermal hypersensitivity to insect bites with the major histocompatibility complex. Firstly, the occurrence of the condition was examined in 304 half-siblings sired by six stallions (A to F). Fourteen cases of dermal hypersensitivity were recognized and all were in the 153 offspring of Stallions C, E and F. Most animals of this group were also investigated for chronic hypersensitivity bronchitis: none of the sires displayed clinical signs of dermal hypersensitivity, but Stallions D, E and F were affected by chronic bronchitis. Among the animals investigated for both conditions only one horse showed coincidence of the two diseases as can be expected when the diseases are not correlated. The frequency of manifest dermal hypersensitivity and/or chronic hypersensitivity bronchitis varied in the half-sibling groups of individual sires. These findings suggest that the allergic conditions are independent entities. Secondly, the occurrence of dermal hypersensitivity was studied in three generations of horses at a stud at which Stallion C had exerted a particularly strong influence. A total of 302 animals, all born and raised at this stud, were surveyed over a period of 12 years. The descendants of Stallion C showed a significantly higher incidence (P less than 0.01) of dermal hypersensitivity (two daughters out of 19; eight second generation offspring out of 103; one third generation offspring out of 85) than did the controls of the same age classes but unrelated to Stallion C at the same stud (0 out of 95).(ABSTRACT TRUNCATED AT 250 WORDS)     

Diagnosis of sarcocystosis in sheep using the indirect fluorescence test and ELISA]

The indirect fluorescent antibody test (IFAT) was compared with the enzyme-linked immunosorbent assay (ELISA) for the detection of specific antibodies to Sarcocystis sp. A set of 275 ovine blood samples was examined by both reactions. Cystozoites of Sarcocystis gigantea were used as the corpuscular antigen for the IFAT. For the diagnostics of sarcocystosis by the ELISA technique used the sandwich test of the antibody titration with a soluble antigen which was also prepared from S. gigantea macrocysts. Our studies confirmed that this antigen did not cross-react with Toxoplasma gondii. Titre 40 was determined as the limit one for the IFAT and titre 80 for the ELISA; which was confirmed by the direct detection of cysts in the muscles (Svobodová, 1989). The results of both methods are shown in Table I. 76.7% of the blood samples reacted positively in the IFAT and 83.6% in the ELISA. These methods were found to be suitable and can be utilized for the intravital routine diagnostics.     

Use of a shrub (Medicago arborea) to control water erosion on steep slopes

Abstract. The effect of Medicago arborea on erosion on a burnt area was studied in experimental plots near Valencia, Spain, between 1989 and 1992. Its growth and development was studied, and its effect was compared with the natural vegetation (matorral) and bare soil. Medicago decreased soil loss by 41.7% and runoff by 25.7% compared with bare soil. However, under natural vegetation soil loss was 27.5% less than under Medicago.     

Studies on mutlispecific resistance of gastrointestinal nematodes to benzimidazoles on dairy-goat farms

Multispecific resistance to benzimidazoles was studied in three selected farms. These farms had bred dairy goats for more than 15 years. The helminths were introduced with the goats at the establishment of the farms which afterwards remained isolated. Nematode resistance could then be related to their own management practices. Faecal egg count tests and egg hatch assays were performed to assess intensity of resistance. The generic (infective larvae in faecal cultures) and specific richness (adult worms) were assessed. The resistant species were Trichostrongylus colubriformis, Teladorsagia circumcincta, Haemonchus contortus and Oesophagostomum venulosum. Faecal egg count reduction tests and egg-hatch assays did not match exactly. Faecal larval counts after treatments gave a distorted picture of multispecific resistance: Haemonchus and Oesophagostomum were very largely over represented. The number of species found in the three farms was relatively low compared with other reports in goat farms of the area. This reduction of diversity might also be due in part to characteristics of breeding management and history (use of permanent pasture and introduction of goats at the establishment of farm).     

Genomic variability among globally distributed isolates of equine arteritis virus.

Equine arteritis virus (EAV), a non-arthropod borne togavirus, has been shown to have a global distribution. To date, no major antigenic variation has been demonstrated between EAV isolates from different geographic origins. In this study, the genomic RNA of EAV isolates obtained from horses of different breeds in various countries around the world was oligonucleotide fingerprinted. Comparisons of these fingerprints were used to determine the extent of genomic variation among such isolates. Comparisons among isolates from North American horses revealed, for the most part, oligonucleotide homologies of less than 60%. Only 29 of the 98 comparisons revealed greater than 60% oligonucleotide homology. Nonetheless, several comparisons indicated a close epidemiologic relationship between isolates from horses of different breeds located in different states. Though all European isolates were of Standardbred origin and were from horses located in northern European countries, the majority had oligonucleotide homologies of less than 60%. Where oligonucleotide homology was apparent, it was, with one exception, greater than 70%. The two isolates from New Zealand had 93.2% oligonucleotide homology. This is indicative of an extremely close epidemiologic relationship. Comparisons between EAV isolates from around the world revealed oligonucleotide homologies between viruses from North America, Europe and New Zealand. In several instances, this homology was greater than 70% and in one case greater than 80%. No oligonucleotide homology was evident in comparisons involving the virus from South Africa. The high level of genomic conservation between certain EAV isolates of disparate geographic origins may reflect dissemination of the virus associated with the international movement of horses. The extent of genomic variation demonstrated between most of the EAV isolates used in this study confirms the need for further investigation of genomic heterogeneity among strains of this virus before techniques that rely upon nucleic acid hybridization can be effectively applied as diagnostic procedures.     

Toxic peripheral neuropathy with demyelination in Sprague-Dawley rats given CGS 21595--a 5-lipoxygenase inhibitor.

Male and female Sprague-Dawley rats were given CGS 21595, a pro-drug that is almost immediately metabolized to CGS 19213, a naphthoquinone that acts as a 5-lipoxygenase inhibitor. The compound was administered by gavage to five groups of Sprague-Dawley rats (group Nos. 1, 5, n = 30; group Nos. 2-4, n = 20) at daily doses of 0, 50, 150, 500, or 1,000 mg/kg for 13 weeks. Rats in the higher dose groups had a reduced weight gain, but significant neurologic signs were not observed. A peripheral neuropathy consisting predominantly of myelin destruction in the spinal nerve roots and sciatic nerves in male rats treated with greater than or equal to 150 mg/kg CGS 21595 and in female rats treated with greater than or equal to 50 mg/kg CGS 21595 for 13 weeks. This lesion was not fully reversible after a recovery period of 4 weeks. Lesions consisted of ballooning of myelin sheaths, infiltration by macrophages, demyelination, and occasional areas of remyelination. Axons were generally preserved, and the brain and spinal cord were not affected. Male and female rats in all treatment groups had cytoplasmic hyaline droplets in the proximal renal tubules. This change was reversible after 4 weeks and was not associated with any other adverse effects on the kidney.     

Cytology of the cerebrospinal fluid in dogs with brain tumors and spinal cord compression. Part 4]

The results of cerebrospinal fluid (CSF) cytology of 9 dogs with brain tumors and 50 dogs with spinal cord compression are discussed. Of the 50 dogs with spinal cord compression, disc protrusion was diagnosed in 31, myelomalacia in 7, discospondylitis in 3 and spinal cord tumors in 9 dogs. In 4 of 9 dogs with brain tumors, tumor cells could be found by the sedimentation apparatus of K?lmel. Pleocytosis existed in 6 patients. In about 70% (29 of 41) of cases with disc protrusion, more than 200 cells could be evaluated in the CSF sediment, consisting mostly of transformed lymphocytes and activated monocytes. As the neurologic deficits increased, the amount of cells and especially cell complexes increased. This was especially evident in cases with myelomalacia of the spinal cord. Only in cases with discospondylitis or spinal cord neoplasia was the CSF cytology unchanged.