Effect of high temperature on infectivity of Toxoplasma gondii tissue cysts in pork

To study the effect of high temperature on infectivity of Toxoplasma gondii tissue cysts, pork from infected pigs was mixed with infected mouse brains and homogenized thoroughly. Twenty-gram samples of infected homogenized meat were sealed in plastic pouches, pressed to a uniform thickness of 2 mm, and subjected to water-bath temperatures of 49, 52, 55, 58, 61, 64, and 67 C for 0.01, 3, 6, 9, 12, 24, 48, and 96 min. Treated samples were digested in HCl-pepsin solution and bioassayed in mice. Toxoplasma gondii tissue cysts remained viable at 52 C for 9.5 min but not for 9.5 min at 58 C; tissue cysts were generally rendered nonviable by heating to 61 C or higher temperature for 3.6 min. Tissue cysts survived once at 64 C for 3 min. These data demonstrate that T. gondii tissue cysts are less heat resistant than encysted Trichinella spiralis larvae.     

Neospora caninum (Apicomplexa) in an aborted equine fetus

Tachyzoites of Neospora caninum were found in sections of lung of an equine fetus aborted 2 mo before term. Individual tachyzoites were approximately 3-5 x 2-3 microns, divided by endodyogeny, and stained positively with anti-N. caninum serum but not with anti-Toxoplasma gondii serum. Toxoplasma gondii antibody was not found in the mare's serum. This is the first report of N. caninum in a horse and indicates that N. caninum can be transmitted transplacentally in equids.     

Enzootic toxoplasmosis in sheep in north-central United States

Of 1,564 serum samples from adult ewes from 33 farms in Iowa, Minnesota, South Dakota, Kansas, and Nebraska where toxoplasmosis-induced ovine abortions had been diagnosed, 65.5% were found positive for Toxoplasma gondii antibodies using the modified agglutination test. Toxoplasma gondii antibody titers of ewes were: less than 64 (34.5%), 64 (14.9%), 256 (22.0%), 1,024 (14.5%), and greater than 4,096 (13.8%). Thus, 28.3% of sheep had high titers (greater than 1,024) indicating recently acquired T. gondii infection. On certain farms, up to 95% of ewes were seropositive. Prevalence of T. gondii antibodies increased with age of the ewe. Of 665 ewes, 53.6% of 1-yr-old ewes were seropositive (titers greater than 64) versus 75% of 5-yr-old ewes. Results indicate that T. gondii infection in sheep in the United States is widespread.     

Evaluation of anti-coccidial drugs' inhibition of Neospora caninum development in cell cultures

Eight anti-coccidial drugs were examined for their efficacies in preventing development of Neospora caninum in bovine monocyte cell cultures. Lasalocid sodium (0.05 microgram/ml), monensin sodium (0.05 microgram/ml), piritrexim (0.01 microgram/ml), pyrimethamine (0.05 microgram/ml), and trimethoprim (5.0 micrograms/ml) were effective in preventing development of intracellular N. caninum tachyzoites (P less than 0.05). No differences (P greater than 0.05) in mean numbers of infected cells compared to controls were observed in cultures treated with amprolium hydrochloride (10.0 micrograms/ml), sulfadiazine (200.0 micrograms/ml), and sulfamethoxazole (200.0 micrograms/ml).     

Transplacental Neospora caninum infection in cats

Transplacental transmission of Neospora caninum was studied in 2 pregnant cats (queens). Queen 1 was inoculated subcutaneously with 2 x 10(6) cell culture-derived N. caninum tachyzoites on day 47 of gestation. She gave birth to a full-term kitten on the 17th day after inoculation. The kitten died the second day after birth due to generalized N. caninum infection. The mother cat was killed on the third day after parturition and was found to have a macerated kitten in the uterus. Severe placentitis, metritis, hepatitis, and nephritis due to N. caninum were seen in tissues from the queen. Queen 2 was fed N. caninum tissue cysts and mated 111 days later. She gave birth to 3 healthy full-term kittens. The kittens were necropsied at 2, 22, and 30 days of age. Neospora caninum was recovered from the organs and was seen in histologic sections in 1 of the 3 kittens. Results indicate that N. caninum can be transplacentally transmitted in cats during acute and chronic stages of infection. Neospora caninum-specific IgG antibodies were demonstrated in the sera of inoculated cats and nursing kittens.     

Culture of rat mesenteric arteriolar smooth muscle cells: effects of platelet-derived growth factor, angiotensin, and nitric oxide on growth

We cultured smooth muscle cells as explants from rat mesenteric arterioles (40–200m in diameter) obtained by injecting a suspension of iron oxide intraarterially and magnetically separating the arterioles after collagenase digestion of adventitial tissue. In third-passaged cells we ascertained smooth muscle purity of >98% by characteristic morphology, contraction responses, and specific immunofluorescence staining. Treatment of growth-arrested (in 0.4% fetal calf serum) cells with platelet-derived growth factor (0.3–7.5 nM) or angiotensin II (0.001–1000 nM) induced ³H-thymidine incorporation and cell proliferation in a dose-dependent manner (P<0.01). S-nitroso-N acetylpencillamine (0.05–0.5 mM), a nitric oxide-generating compound, inhibited 10% fetal calf serum-induced ³H-thymidine incorporation (P<0.05) and cell proliferation (P<0.01). The antimitogenic effect of S-nitroso-N-acetylpencillamine was significantly reduced by hemoglobin and potentiated by superoxide dismutase (P<0.01). In addition to a new technique for culturing mesenteric arteriolar smooth muscle cells, these findings provide evidence that platelet-derived growth factor, angiotensin II, and nitric oxide may be involved in their growth control.     

Potential use of the essential oil ofTrachyspermum ammi against seed-borne fungi of Guar (Cyamopsis tetragonoloba L. (Taub.))

Essential oils isolated from leaves and seeds of seven umbelliferous plants were tested against the growth ofAspergillus flavus. Those from seeds ofTrachyspermum ammi, Cuminum cyminum, Carum carvi, Daucus carota and from leaves ofAnethum graveolens exhibited antifungal activity against the test fungus. Amongst these, oil from seeds ofTrachyspermum ammi was most toxic. Its minimum inhibitory concentration was 300 ppm, at which it exhibited fungistatic but not phytotoxic properties, when tested at 200, 300 and 400 ppm. The fungitoxic potency ofTrachyspermum seed oil remained unchanged after a long storage period and at high inoculum density of the test fungus. The oil was thermostable and was more efficaceous than the fungicides Agrosan G.N., Benlate, Ceresan, Dithane M-45 and Thiovit commonly used for the control of plant diseases.     

Identification of a UDP-glucosyltransferase conferring deoxynivalenol resistance in Aegilops tauschii and wheat

Aegilops tauschii is the diploid progenitor of the wheat D subgenome and a valuable resource for wheat breeding, yet, genetic analysis of resistance against Fusarium head blight (FHB) and the major Fusarium mycotoxin deoxynivalenol (DON) is lacking. We treated a panel of 147 Ae. tauschii accessions with either Fusarium graminearum spores or DON solution and recorded the associated disease spread or toxin-induced bleaching. A k-mer-based association mapping pipeline dissected the genetic basis of resistance and identified candidate genes. After DON infiltration nine accessions revealed severe bleaching symptoms concomitant with lower conversion rates of DON into the non-toxic DON-3-O-glucoside. We identified the gene AET5Gv20385300 on chromosome 5D encoding a uridine diphosphate (UDP)-glucosyltransferase (UGT) as the causal variant and the mutant allele resulting in a truncated protein was only found in the nine susceptible accessions. This UGT is also polymorphic in hexaploid wheat and when expressed in Saccharomyces cerevisiae only the full-length gene conferred resistance against DON. Analysing the D subgenome helped to elucidate the genetic control of FHB resistance and identified a UGT involved in DON detoxification in Ae. tauschii and hexaploid wheat. This resistance mechanism is highly conserved since the UGT is orthologous to the barley UGT HvUGT13248 indicating descent from a common ancestor of wheat and barley.     

Lipase-catalysed esterification reaction in a tubular reactor

Summary Lipozyme catalysed esterification of oleic acid with lauryl alcohol was studied in a tubular recycle reactor. Water formed during the reaction was removed by adsorption, either by passing the product mixture through another column packed with molecular sieve, or by packing the reactor partially with silica gel.