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.     

Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins

The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID₅₀) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID₅₀ per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml⁻¹ of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml⁻¹ of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.     

Eastern equine encephalitis in a flock of emus (Dromaius novaehollandiae).

Eastern equine encephalitis (EEE) was diagnosed in a flock of emus in southeastern Louisiana. The outbreak involved juvenile and adult breeders ranging in age from 20 to 36 months, with an attack rate of 76% and a case fatality rate of 87%. The diagnosis was confirmed by isolation and characterization of the viral agent, and by detection of EEE antibody in two recovered emus. High mortality was preceded by marked depression, hemorrhagic diarrhea, and emesis of blood-stained ingesta. On postmortem examination, hemorrhagic enteritis and multiple petechia of viscera were observed. Microscopic changes included severe necrosis of hepatocytes, intestinal mucosa, and necrotizing vasculitis of the spleen and lamina propria of the intestine. No nervous system lesions were observed. This outbreak occurred concurrently with EEE in horses and was attributed to unseasonably heavy rainfall with an abundance of arthropod vectors and proximity to free-living reservoir host species.     

Chronic diarrhea associated with intestinal anomalies in a six-year-old dog.

A 6-year-old mixed-breed dog was referred because of chronic diarrhea, which had begun after antibiotic treatment for pulmonary disease. Intestinal bacterial overgrowth was presumptively diagnosed on the basis of intestinal mucosal cytologic examination and low serum cobalamin concentration. When the diarrhea did not respond to treatment, contrast radiography was performed and a short small intestine was found. The diarrhea could not be managed by medical treatment, and the dog was euthanatized. Necropsy revealed 2 blind intestinal pouches attached to a 46-cm length of small intestine connecting the stomach and colon. The dog's deteriorating condition was attributable to necrosis and rupture of 1 of the pouches.     

Anesthesia for patients with severe systemic debilitation.

Any patient presenting for anesthesia and surgery who has a severe debilitating problem is at great risk for a prolonged recovery, renal failure, or hypotension. A stable hemodynamic state should be maintained. Fluid homeostasis should be ensured. Anesthetic induction techniques using narcotics and benzodiazepines are my preferred approach, with maintenance of anesthesia incorporating isoflurane, narcotics, or both or the use of regional anesthesia.     

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.     

Cardiopathological observation on a case of persistent ventricular tachycardia in a pony mare.

A 20-year-old pony mare with persistent ventricular tachycardia (VT) was examined cardiopathologically. At necropsy, the heart was enlarged and rounded with both ventricular dilatation. In a longitudinal section of the ventricular septum, a large grayish white patchy lesion (5 x 25 mm) was detected in the relatively higher portion. Microscopically, the lesion was extensive myocardial fibrosis located in the vicinity of the proximal part of the left bundle branch. Partially the fibrotic lesion was in contact with the branch. Such a lesion might play an important role in creating a suitable background for the development of VT via automaticity or reentry mechanism.