Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation.

Serum-free mouse embryo (SFME) cells, derived in medium in which serum is replaced with growth factors and other supplements, are proastroblasts that are acutely dependent on epidermal growth factor (EGF) for survival. Ultrastructurally, an early change found in SFME cells deprived of EGF was a loss of polysomes which sedimentation analysis confirmed to be a shift from polysomes to monosomes. The ribosomal shift was not accompanied by decreased steady-state level of cytoplasmic actin mRNA examined as an indicator of cellular mRNA level. With time the cells became small and severely degenerate and exhibited nuclear morphology characteristic of apoptosis. Genomic DNA isolated from cultures undergoing EGF deprivation-dependent cell death exhibited a pattern of fragmentation resulting from endonuclease activation characteristic of cells undergoing apoptosis or programmed cell death. Flow cytometric analysis indicated that cultures in the absence of EGF contained almost exclusively G1-phase cells. Some of the phenomena associated with EGF deprivation of SFME cells are similar to those observed upon NGF deprivation of nerve cells in culture, suggesting that these neuroectodermal-derived cell types share common mechanisms of proliferative control involving peptide growth factor-dependent survival.

Effect of vitamin B12 on performance and tissue selenium content in rats fed sub-toxic levels of selenite.

The effects of vitamin B12 status on growth and tissue selenium distribution were studied in Sprague-Dawley rats chronically exposed to subtoxic levels of selenite. Vitamin B12 status was monitored by urinary methylmalonic acid excretion and by liver and plasma vitamin B12 levels. Selenite absorption was unaffected by dietary level of vitamin B12. A significant (P < 0.05) interaction of vitamin B12 and selenium was found on growth of rats fed vitamin B12 deficient or control diets. In vitamin B12 depleted rats, there were significant histologic changes in the liver that were characterized by micronodules and regeneration, bile duct reduplication, mild cirrhosis, necrosis of individual hepatocytes and other minor histologic changes. There was no gross or histologic evidence of liver toxicity in rats supplemented with vitamin B12. Rats pair-fed 9 mg/kg selenium with vitamin B12 had significantly lower liver and kidney selenium levels and significantly higher blood selenium levels compared to rats fed the diet without vitamin B12. These results are consistent with the hypothesis that vitamin B12 deficiency limits selenium methylation and excretion, resulting in higher tissue selenium levels and subsequent toxicity.

Fibroblast growth factor inhibits expression of neural markers in cultures of zebrafish early embryo cells.

Basic fibroblast growth factor (FGF) regulation of developmental markers in cell cultures derived from early zebrafish embryos was examined with the goal of in vitro culture of zebrafish embryonic stem cells and gaining an understanding of extracellular influences on early embryonic development. Markers were stem/primordial germ cell markers pou-2 and vas, neural markers zp-50, pax[zf-a], en-3, and wnt-1, and mesodermal markers gsc and myoD. Previously we had shown that FGF prevents the development of zebrafish pigment cells in vitro. In our culture system, FGF reduced expression of neural-specific markers, possibly implicating the FGF family in suppression of early neural cell development. Exposure to FGF for 24 hours at the time of seeding the cells was sufficient to suppress neural marker expression for a subsequent 4 days of culture, while absence of FGF for the first 24 hours of culture nullified the effect of FGF added subsequently. FGF predictably increased expression of gsc and myoD. Vas expression was unaffected, while pou-2 expression decreased with time in culture in the presence or absence of FGF. However, in situ hybridization identified a subpopulation of cells expressing pou-2, suggesting the possible continued existence of undifferentiated stem cells in the cultures.

Measurement of IgG concentration in ovine fetal fluids: a useful diagnostic test.

The Veterinary Diagnostic Laboratory at Oregon State University received 172 aborted ovine fetuses during the 1985-1987 lambing seasons; from 120 of these, body fluids were evaluated for IgG levels. Fifty-two (43%) of the fetal fluids had immunoglobulin G (IgG) levels greater than 15 mg/dl. Forty-five (87%) of the fluids with elevated IgG levels were confirmed or presumed toxoplasma or chlamydia abortions. A mean fetal fluid IgG concentration of 111.5 +/- 78 mg/dl was found for the 26 toxoplasma abortions; for the 19 chlamydia abortions, a mean IgG concentration of 109 +/- 91 mg/dl was found. Antibody titers equal to or greater than 1:40 against Toxoplasma gondii were detected in 23 fetal fluids. Fetal fluid IgG concentration less than 15 mg/dl was found to be associated with bacterial organisms (i.e., Campylobacter sp.) as the confirmed or presumed cause of abortion. These results suggest that measurement of fetal fluid IgG concentration is a useful, supportive diagnostic test in determining the cause of ovine abortion, and should be included as a routine laboratory procedure for ovine abortion diagnosis.

Identification of ferrous sulfate toxicity in a commercial broiler flock.

A 2.5% mortality rate was observed in a flock of 19,000 commercial one-day-old broiler chicks that had been placed 24 hours previously on litter treated with ferrous sulfate heptahydrate. Ulcerative ventriculitis and severe hepatopathy were the primary lesions observed grossly and microscopically. Pooled digesta contained 6854 ppm iron. Lesions identical to those found in the field case were reproduced experimentally.

Toxicity of excess dicalcium phosphate in the diet of turkey poults.

High mortality in 1-week-old commercial turkey poults was attributed to the accidental substitution of dicalcium phosphate in the diet for corn. At necropsy of the affected birds, lesions were observed in the upper gastrointestinal tract, suggesting acute ingestion of a caustic substance. Mortality and gross and microscopic lesions were reproduced experimentally in turkey poults fed diets similar to the diet fed in the field case. The cause of these lesions was attributed to increased pH due to the phosphoric acid content of the diet.

Intestinal helminths of river otters (Lutra canadensis) from the Pacific northwest.

The intestinal helminth fauna of river otters, Lutra canadensis, from the Pacific Northwest was characterized by low species richness and intensity of infection. River otters from the lower Columbia River (n = 23) were infected with 9 species of helminths (83% prevalence); those from a relatively undisturbed reference area near the headwaters of the Trask and Wilson rivers on the Oregon coast (n = 6) were infected by 5 species of helminths (100% prevalence). Single species of Eucestoda (Schistocephalus solidus), Digenea (Euparyphium inerme), Acanthocephala (Corynosoma strumosum), and 8 species of Nematoda (Strongyloides lutrae; larvae of Eustrongylides sp., Anisakis sp., and Contracaecum sp.; 3 of Cystidicolidae, and Hedruris sp.) were collected. Most species are typical of piscine definitive hosts and were present as incidental parasites of river otters. Notably, specimens of Euparyphium inerme are reported for the first time in river otters from North America; occurrence of other helminths constitutes new host or geographic records for parasites in river otters in Oregon and Washington. Parasites with marine life cycles were acquired by river otters in freshwater habitats at a great distance from the ocean. The helminth fauna of river otters in the Pacific Northwest was influenced primarily by ecological factors and was indicative of eclectic food habits and the relatively extensive home ranges occupied by these mustelids.

Culture of cells from juvenile worms of Schistosoma mansoni.

Tissue disruption methods were developed and serum-free cell culture media formulated for the maintenance in vitro of cells from juvenile worms (day 18 after infection) of Schistosoma mansoni. Cultures maintained viability for up to 6 mo when plated on a feeder layer of irradiated rat liver cells and survived primarily as clusters of small (2.5-4 microns diameter) cells with a high nuclear-to-cytoplasmic ratio and relatively few organelles identified by electron microscopy. Cultures synthesized a protein profile similar to that of intact worms, and the cell clusters maintained a time- and concentration-dependent contractile response to serotonin. Cells synthesizing DNA were detected by precursor incorporation and flow cytometry in cultures initially and also after several weeks in vitro, although the percentage of cells synthesizing DNA decreased with time. Efforts to identify peptide growth factor-responsive tyrosine phosphorylation were negative, and the overall amount of S. mansoni phosphotyrosine-containing proteins identified by western blot with anti-phosphotyrosine monoclonal antibody was much less than that found in a peptide growth factor-responsive mouse cell line.

Megaesophagus in 15 llamas: a retrospective study (1985-1993).

Megaesophagus was diagnosed in 15 llamas on the basis of survey and contrast radiography. Age of onset ranged from 13 months to 9.5 years. Clinical signs varied, with salivation being the most common; regurgitation or dysphagia were noted in only 33% of the cases. Duration of the disorder ranged from 1 week to 5 years. Organophosphate toxicity was the cause of megaesophagus in 1 llama. While most cases were of unknown etiology, the 3 with histological abnormalities included 1 with vagal neuropathy and 2 with a degenerative myopathy of esophageal muscles. The causes in most were undetermined.

Effect of dietary fluoride on selenite toxicity in the rat.

Three factorial experiments were conducted to determine if high dietary fluoride (F) would inhibit selenite toxicity in rats. Initially, three levels of selenite (0.05, 3, and 5 mg/kg diet) were matched against three levels of F (2, 75, and 150 mg/kg diet). Fluoride failed to prevent the depressive effect of selenite on 8-wk food intake and body wt gain. Selenium (Se) concentration of plasma and kidney and enzymatic activity of whole blood glutathione peroxidase (GSH-Px) were also unaffected by F. Liver Se concentration, however, was slightly (12%) but significantly (p < 0.025) reduced when the highest F and Se levels were combined. Fluoride (150 mg/kg) appeared to reduce liver selenite toxicity (5 mg/kg). Therefore, further study focused on liver histology with treatments that eliminated the middle levels of selenite and F. Fluoride prevented the hepatic necrosis seen in selenite-toxic rats. Similar histological lesions were not observed for kidney or heart. Fluoride partially (26%) but significantly (p < 0.025) reduced thiobarbituric-reactive substances in selenite-toxic rats, but there was no F effect on intracellular distribution of liver Se, glutathione levels in liver and kidney, or on liver xanthine oxidase activity. Overall, the protective effect of F on selenite toxicity appears to be confined to liver pathology. The exact mechanism for this effect, however, remains unclear.

Safety and efficacy of two sustained-release intrareticular selenium supplements and the associated placental and colostral transfer of selenium in beef cattle.

One hundred fifty Se-deficient, pregnant, crossbred beef cows were assigned to 1 of 4 treatment groups: group A, Se-deficient control; group B, 1 Se bolus at 0 and 119 days; group C, 1 Se bolus at 0 days; and group D, 2 Se pellets at 0 days. The Se bolus is an osmotic pump designed to release 3 mg of Se/d into the reticulorumen. The Se pellets weight approximately 30 g and contain 10% elemental Se, which is liberated in the reticulorumen. The Se bolus is designed to provide Se supplementation for 120 days and the Se pellets provide supplementation for up to 18 months. Cattle were maintained on Se-deficient pasture or forages prepared from these pastures for the duration of the experiment. Blood samples were collected from cows prior to treatment (time 0) and at 28, 52, 119, and 220 days thereafter and analyzed for blood Se (BSe) concentration. Body weights were recorded at each sampling time. Blood Se concentration of cows from all supplemented groups were significantly (P less than 0.01) higher than control values at all sample dates after treatments began. By the end of the 220-day study, treatment group-B cattle had significantly (P less than 0.01) higher BSe concentrations than any other group. Body weights of treatment groups fluctuated throughout the study, but did not differ (P greater than 0.05) between groups. One cow and 6 calves born to cows during the experimental period died. Necropsy of 5 calves provided no evidence linking these deaths to treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Enteric coccidia (Apicomplexa) in the small intestine of the northern spotted owl (Strix occidentalis caurina).

Sporulated oocysts (mean dimensions = 13.0 x 10.8 microns) and sporocysts (11.3 x 5.5 microns) of a coccidian resembling Frenkelia sp. or Sarcocystis sp. were present in the lamina propria of the small intestine of a naturally-infected northern spotted owl (Strix occidentalis caurina) collected near Medford, Oregon (USA). Dimensions of these oocytes and sporocysts appear to be considerably smaller than those from other sarcocystid species with avian definitive hosts. Additionally, numerous developmental stages and unsporulated oocysts (mean dimensions 22.8 x 17.8 microns) of a possible species of Isospora also were observed in the intestinal epithelium. This constitutes the first report of enteric coccidia from spotted owls. Neither parasite appeared to cause the death of the bird.

Helminth parasites of northern spotted owls (Strix occidentalis caurina) from Oregon.

Helminth parasites are reported for the first time from northern spotted owls. Seventy-one percent of a sample of Strix occidentalis caurina from western Oregon was infected. Nematodes (Porrocaecum depressum, Capillaria falconis, Microtetrameres sp. and Synhimantus hamatus) were the most prevalent parasites although cestodes (Paruterina rauschi) and acanthocephalans (Centrorhynchus conspectus) were also represented. There was an association between components of this helminth fauna and the diet of spotted owls which is dominated by small rodents. The occurrence of P. rauschi rather than P. candelabraria in this geographic region and host-species may provide additional support for recognition of a parapatric distribution in the ranges of Paruterina spp. among strigiforms in the Nearctic.

Hemorrhagic enteritis caused by Clostridium perfringens type C in a foal.

A 4-day-old foal died with bloody diarrhea. Using a mouse neutralization test, Clostridium perfringens type C was isolated from intestinal contents, and alpha and beta toxins were identified. About 4 m of the jejunum had severe necrohemorrhagic enteritis. Microscopically, large, rod-shaped, gram-positive bacteria were seen on necrotic intestinal villi by use of Brown and Hopp's stains.

Hepatic cirrhosis and hemochromatosis in three horses.

Hemochromatosis, an iron storage disease, was diagnosed in 3 horses with hepatic cirrhosis. Each horse had bridging portal fibrosis and abundant iron deposits in the cytoplasm of hepatocytes. Serum concentrations of liver-derived enzymes and total bile acids were high. However, serum iron concentration was not high, and iron binding capacity was only 46% saturated in the 1 horse in which it was measured. However, the concentration of iron in the liver of this horse was 20 times the reference limits. Hemochromatosis is common in mynah birds and human beings. There are several types of this iron storage disease. In human beings, there is a familial disease with iron absorption inappropriate for the level of stored iron. Hemochromatosis also occurs secondary to cirrhosis of the liver, and in certain anemia diseases. Another type of hemochromatosis is seen in association with dietary iron overload. These horses were not related, and there was not any evidence of high dietary iron intake.

Pleuritis and pneumonia attributed to a conifer twig in a bronchus of a horse.

A conifer twig was responsible for severe fibrinopyogranulomatous adhesive pleuritis and pneumonia in a horse. At necropsy, the twig was found in a terminal bronchus and extended into the lung parenchyma, through the pleura, and into the accumulated exudate. Inhalation of plant material may be a more common cause of pleuritis than previously recognized. Meticulous examination at necropsy would be necessary to make the diagnosis.

Photosensitization associated with exposure to Pithomyces chartarum in lambs.

An epidemic of photosensitization was observed in a group of lambs on irrigated autumn pasture in western Oregon. Signs included crusting, necrosis, and sloughing of the skin over the nostrils, lips, and ears, and of the mucous membranes of the buccal regions. Microscopic examination of plant material from the pasture disclosed spores of Pithomyces chartarum. This fungus has been documented as a causal factor in photosensitization in sheep and cattle (facial eczema) in other parts of the world. An infective agent or other plant material that could have induced the clinical signs in the lambs was not evident. Weather and humidity conditions were ideal for fungal growth during the grazing period, and the fungus was detected in large numbers before and during the epidemic. Even though facial eczema has not been reported previously in northwestern United States, we feel the circumstances surrounding this epidemic warrant such a diagnosis.

Efficacy of a vaccine to prevent Chlamydia- or Campylobacter-induced abortions in ewes.

In a sheep flock, Chlamydia psittaci, Campylobacter fetus, Ca jejuni, and Salmonella dublin caused abortions. A vaccine that contained C psittaci type I from 2 sources: a cow with pneumonia and an aborted ovine fetus, Ca fetus, Ca jejuni, and 4 strains of K99 Escherichia coli was given to 240 ewes before they were bred. All fetuses, placentas, and lambs, that died within 36 hours of birth were examined for infectious agents. Of 55 abortions, 30 (55%) were caused by Chlamydia or Campylobacter spp; 25 of the 30 (83%) abortions took place in the nonvaccinated group (n = 240). Forty-five more lambs survived in the vaccinated group than in the nonvaccinated group. Abortion rates for Chlamydia and Campylobacter spp (2.1 vs 10.4% in vaccinated and nonvaccinated groups, respectively) were significantly different (P = 0.003). Abortion rates for S dublin were not significantly different between groups. The Salmonella epizootic was controlled quickly by sanitation and treatment procedures. The vaccine was at least 80% efficacious against Chlamydia and Campylobacter spp and appeared to be protective.

Serologic and histologic diagnosis of toxoplasmic abortions in sheep in Oregon.

Between 1983 and 1989, we examined 586 fetuses and dead lambs submitted to a diagnostic laboratory in Oregon. Toxoplasmosis was diagnosed in 74 (12.6%), campylobacteriosis in 89 (15.2%), chlamydiosis in 75 (12.8%), miscellaneous infections in 153 (26.1%), noninfectious causes in 37 (6.3%), and undetermined causes in 158 (27.0%). Detection of Toxoplasma gondii agglutinating antibodies (greater than or equal to 20) in body fluids or heart serum was considered specific for the diagnosis of toxoplasmosis in ovine fetuses or lambs. Toxoplasma gondii antibody titers (reciprocal) in 285 fluids from fetuses or dead lambs were less than 20 (64.5%), 20 (4.5%), 40 (1.7%), 80 (0.7%), 160 (2.1%), 320 (1.7%), 640 (4.9%), 1,280 (2.8%), 2,560 (4.5%), 5,120 (3.8%), 10,240 (2.1%), and greater than or equal to 20,480 (6.3%).