Inhibitory effect of penicillin on caecal urease activity in chickens fed on a low protein diet plus urea

1. The effects of dietary penicillin on the urease activities of small intestine, large intestine, caecum and their contents, liver and kidney in chickens fed a diet containing 5 g/kg protein plus urea were examined.

2. About 0.88 of the total urease activity determined was observed in intestinal contents, of which 0.95 of the activity was accounted for by caecal contents, 0.05 by colo‐rectal contents and none by small intestinal contents. Intestinal tissues (caecum included), liver and kidney accounted for 0.03, 0.06 and 0.02, respectively, of the total urease activity.

3. Dietary penicillin decreased urease activity to 0.17 in caecal contents and to 0.05 in colo‐rectal contents of the corresponding control values (P< 0.01). The urease activity of caecal tissue was lowered by penicillin to half that of control activity (P < 0.05) but none of the activities of other tissues were affected.

4. It is concluded that, even when the urease activity is stimulated by dietary urea, 20 mg/kg dietary penicillin can strikingly lower it in the caecum, where most of the urease activity in the chicken body is to be found.     

Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.     

Ammonium nitrogen in soil solution and seed nitrogen affect the percentage of establishment of rice plants in submerged soil

The effect of ammonium nitrogen concentration in soil solution on the establishment of rice plants was examined. The increase of the concentration decreased the percentage of establishment of seeds sown in submerged soil, although most of seeds sown on submerged soil became established. Therefore, the increase of ammonium nitrogen concentration in soil solution may impair the establishment of seeds sown in submerged soil, which would occur presumably because the increase delays the spear growth and emergence without the decline of soil redox potential. Several seed lots with various nitrogen contents were obtained from rice plants grown under various conditions. The percentage of establishment of low-nitrogen seeds sown in submerged soil was much lower than that of high-nitrogen seeds, especially in soils whose solution contained a large amount of ammonium nitrogen. However, the difference in the percentage of establishment between high- and low-nitrogen seeds sown on submerged soil was much smaller. Therefore, the increase of seed nitrogen content may improve the percentage of establishment of seeds sown in submerged soil, presumably because the increase accelerates the spear growth and emergence. Consequently, for direct sowing in submerged soil, ammonium nitrogen concentration in soil solution should remain low because ammonium nitrogen exerts an adverse effect on seedling establishment, and vigorous seeds with a high nitrogen content should be sown because seed nitrogen exerts a beneficial effect on seedling establishment.     

Comparison of the effects of seed coating with tungsten and molybdenum compounds on seedling establishment rates of rice,wheat, barley,and soybean under flooded conditions

Seed coating with molybdenum compounds improves seedling establishment for rice, wheat, barley, and soybean when such seeds were sown under flooded conditions. Tungsten belongs to the same chemical group as molybdenum in the periodic table, and similar to molybdenum, inhibits the generation of sulfide ions. Here, the effects of tungsten and molybdenum containing seed coatings on seedling establishment under flooded conditions were compared using rice, wheat, barley, and soybean. In rice, the effects of tungsten compounds on seedling establishment varied. Tungsten trioxide had little effect but tungstic acid and ammonium phosphotungstate significantly improved seedling establishment when the amounts were at least .1–.2 mol W kg^?1. Although the effect of tungsten coating varied depending on the compound used, ammonium phosphotungstate, along with other tungsten compounds, improved seedling establishment in a manner comparable with that of molybdenum compounds. For wheat and barley, ammonium phosphotungstate treatment resulted in a significant increase in establishment that was only slightly less than the results observed using molybdenum compounds. Tungstic acid and ammonium phosphotungstate treatments improved soybean establishment in a significant manner that was comparable with those of molybdenum compounds. Collectively, these results suggest that tungsten compounds, as well as molybdenum compounds, improve seedling establishment under flooded conditions.     

Changes in organosulfur compounds in garlic cloves during storage

We determined the changes in the contents of three gamma-glutamyl peptides and four sulfoxides in garlic cloves during storage at -3, 4, and 23 degrees C for 150 days using a validated high-performance liquid chromatography method that we reported recently. When garlic was stored at 4 degrees C for 150 days, marked conversion of the gamma-glutamyl peptides, gamma-L-glutamyl-S-allyl-L-cysteine and gamma-L-glutamyl-S-(trans-1-propenyl)-L-cysteine (GSPC), to sulfoxides, alliin and isoalliin, was observed. Interestingly, however, when garlic was stored at 23 degrees C, a decrease in GSPC and a marked increase in cycloalliin, rather than isoalliin, occurred. To elucidate in detail the mechanism involved, the conversion of isoalliin to cycloalliin in both buffer solutions (pH 4.6, 5.5, and 6.5) and garlic cloves at 25 and 35 degrees C was examined. Decreases in the concentration of isoalliin in both the solutions and the garlic cloves during storage followed first-order kinetics and coincided with the conversion of cycloalliin. Our data indicated that isoalliin produced enzymatically from GSPC is chemically converted to cycloalliin and that the cycloalliin content of garlic cloves increases during storage at higher temperature. These data may be useful for controlling the quality and biological activities of garlic and its preparations.