Dietary phytase increases the true absorption and endogenous fecal excretion of zinc in growing pigs given a corn-soybean meal based diet

We investigated the effect of dietary phytase on the true absorption and endogenous fecal excretion of zinc (Zn) using ⁶⁷Zn in growing pigs given a corn-soybean meal based diet. Ten crossbred barrows were fed the control diet containing 90-mg/kg Zn, 2.3-g/kg phytate-phosphorus and 3.7-g/kg non-phytate-phosphorus or the phytase diet containing similar amounts of Zn and phytate-phosphorus, and 1.4-g/kg non-phytate-phosphorus with 750-PU/kg phytase for 12 h/day. On day 6, the pigs were given 200 g of the corresponding diet labeled by ⁶⁷Zn for 2 h. We measured feed intake, fecal Zn concentration and ⁶⁷Zn abundance for the determination of apparent absorption, true absorption and endogenous fecal excretion of Zn. Although the apparent absorption of Zn did not significantly differ between the dietary groups, the phytase group had significantly more ( P  < 0.05) true absorption of Zn than the control group. The endogenous fecal excretion of Zn tended to be more ( P  = 0.07) in the phytase group than in the control group. These results suggest that dietary phytase improves Zn bioavailability through increasing the true absorption of Zn in growing pigs, which results in stimulating the endogenous fecal excretion of Zn when dietary Zn satisfies its requirement.     

Brewer's yeast efficiently degrades phytate phosphorus in a corn-soybean meal diet during soaking treatment

Microbes such as yeast and Aspergillus are known to produce phytase, and Aspergillus phytase has been used as a feed additive for improving phytate-phosphorus bioavailability in monogastric animals. We measured phytase activity in some by-products from fermented food and beverage productions by yeast and Aspergillus . The phytase activity was as high as 3577 and 2225 PU/kg DM in raw and dried brewer's yeasts, respectively. On the other hand, the phytase activity was approximately 400 PU/kg DM in white-wine yeast and red-wine yeast. The phytase activity was further low in natto (fermented soybean) residue, soy sauce cake, rice brewer's grain and the activity was not detected in dried corn-barley distiller's grain with soluble and sweet-potato distiller's residue. The stability of phytase against pepsin was much lower in the brewer's yeast than in an Aspergillus phytase preparation. On the other hand, the addition of raw brewer's yeast effectively degraded phytate phosphorus in a corn-soybean meal diet during soaking. These results suggest that phytase in the examined by-products is not suitable for the phytase source of conventional diets, but that the soaking treatment with a raw brewer's yeast is an alternative method for improving phytate-phosphorus bioavailability in corn-soybean meal diets for pigs.     

Comparison of electrophoretic patterns of soluble proteins and isozymes of the red rot disease fungus Pythium sp. isolated from Porphyra yezoensis from Korea and Japan

SUMMARY: Pythium sp., the causative organism of red rot disease, isolated from Porphyra yezoensis from Wando in Korea was compared with those from Miyagi, Aichi, and Fukuoka Prefectures by means of electrophoretic patterns of soluble proteins and isozymes. Native and sodium dodecylsulfate–polyacrylamide gel electrophoretic patterns of soluble proteins revealed small but significant differences among the four isolates. Among the 14 detected enzymes, catalase (CAT), carboxylesterase (CE), glucose-6-phosphate dehydrogenase (G6PD), glucosephosphate isomerase (GPI), isocitrate dehydrogenase (IDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and xanthine dehydrogenase (XDH) possessed isozymes. The dissimilarity value of isozyme banding patterns for four isolates by unweighted pair-group method using arithmetic averages (UPGMA) cluster analysis was highest (0.51) between the isolate from Fukuoka and those from Miyagi and Aichi, and lowest (0.19) between the isolate from Miyagi and that from Aichi. Cluster analysis of data from isozyme banding patterns grouped the isolates from Miyagi and Aichi. These results indicate that the isolates from Miyagi and Aichi are similar, but each isolate from Wando and Fukuoka differed slightly from those of Miyagi and Aichi.     

Clinicopathological survey of 101 canine mammary gland tumors: differences between small-breed dogs and others

Clinicopathological features of mammary gland tumors (MGTs) in 101 dogs were evaluated retrospectively. The incidence of histological malignancy in 60 small- and 41 other-breed dogs were 25% and 58.5%, respectively. In 82 epithelial MGTs, small-sized tumors (< 3 cm) or non-invasive tumors were predominant in small breeds. In multivariate survival analysis, small breed (p=0.048) and lower stage of tumor cell invasion (p=0.006) were significantly associated with longer survival time. These results suggest that the incidence of histological or biological malignancy in MGTs is lower in small-breed dogs than in others.     

Characterization and differentiation of <Emphasis Type="Italic">Erwinia carotovora</Emphasis> strains from mulberry trees

We recently reported that two diverse types (types 1 and 2) were identified among strains of Erwinia carotovora from mulberry trees. Type 1 strains were similar to E. carotovora subsp. carotovora (Ecc), whereas type 2 strains were distinct from Ecc and other E. carotovora strains. In this study, seven more mulberry strains of type 2 and reference strains were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified of polymorphic DNA (RAPD). On the basis of SDS-PAGE profiles of whole-cell proteins, type 2 strains had high similarity with one another. In addition, they had an unique peptide band with a molecular mass of approximately 28kDa. RAPD analysis showed that they were also effectively differentiated by a strong, specific RAPD fragment for type 2 strains. Based on these two approaches, we have confirmed that the present type 2 strains from mulberry can be discriminated clearly from other soft rot Erwinia species.     

Antigen-specific histamine release in dogs with food hypersensitivity

An in vitro evidence of IgE-mediated hypersensitivity to food allergens was detected by positive results of antigen-specific histamine release in dogs with food hypersensitivity. Eight dogs were diagnosed to have food hypersensitivity based on identification of offending food allergens with food elimination followed by oral food provocation. The percentages of histamine release against the stimulation of offending food allergens in the cases ranged from 2.1% to 70.9%. Six of the 8 cases showed histamine release higher than those of healthy control dogs. Four dogs showed relatively high histamine release at the percentage beyond 10% that was compatible with a positive value of histamine release in humans with food hypersensitivity. These findings would suggest that IgE-mediated hypersensitivity against food allergens could be involved in canine food hypersensitivity.     

Lesional expression of thymus and activation-regulated chemokine in canine atopic dermatitis

Glucocorticoids are reported to bias cytokines to a Th2 phenotype. However, this dogma has been advanced largely from studies utilizing potent glucocorticoid analogs. The current study was conducted to revisit the issue of glucocorticoid modulation of Th1/Th2 cytokine production and evaluate migration inhibitory factor (MIF) mRNA expression in cultured pig splenocytes treated with physiologically relevant concentrations of cortisol (CORT). Dexamethasone (DEX) was included for comparison. In Experiment 1, DEX, at 150 and 300 nM, suppressed concanavalin (ConA)-stimulated IFNgamma at both 12 and 24 h in culture, and IL-10 at 24h (P<0.05). Both 150 and 300 nM CORT suppressed IL-10 at 24 h (P<0.05), but neither concentration affected IFNgamma at 24 h. In Experiment 2, cells were cultured with a broader range of CORT for 48 h following ConA. Parallel cultures with identical treatments also were conducted in separate plates for evaluation of glucocorticoid regulation of MIF mRNA. IFNgamma was reduced by 300 nM DEX at 12, 24, and 48 h (P<0.05), whereas 150 and 300 nM CORT blunted IFNgamma at 24 h (P<0.05), but not 48 h. ConA increased IL-2 (P<0.01), but none of the steroid treatments affected IL-2. At both 12 and 24 h, IL-10 was reduced by 300 nM DEX and by 150 and 300 nM CORT (P<0.05). ConA increased relative abundance of MIF mRNA (P<0.001), but no steroid treatment affected MIF mRNA. In Experiment 3, steroid additions were delayed by 24 h after ConA, and cytokine concentrations evaluated 48 h later. Again, separate cultures were used for determination of effect of treatments on MIF mRNA. None of the steroid treatments affected IFNgamma, but 300 nM DEX reduced IL-10 (P<0.05). All of the CORT treatments (75-300 nM) reduced MIF mRNA (P<0.05), whereas DEX did not affect MIF mRNA in this experiment. The current experiments suggest that both DEX and high physiological concentrations of CORT can suppress both type 1 and type 2-like cytokines in cultured pig splenocytes. But, IL-10 was generally more sensitive to CORT suppression with increased time in culture than was IFNgamma. In addition, MIF mRNA could be suppressed by delayed addition of CORT to porcine splenocytes. Taken together, the data do not support the hypothesis that CORT directs the cytokine milieu toward a type 2 bias in cultured pig splenocytes.     

Proliferation of canine intervertebral disk chondrocytes in three-dimensional alginate microsphere culture

Proliferation of chondrocytes from nucleus pulposus (NP) and anulus fibrosus (AF) was confirmed in three-dimensional culture using alginate microspheres. Cells isolated from NP and AF were incorporated in microspheres and cultured for 14 days. Round mononuclear cells of 20-25 microm in diameter proliferated and formed aggregates. At day 14, alcian blue positive matrix surrounded the proliferating cells. The cells had cytoplasmic vacuoles stained positively by toluidine blue. On electron microscopy, the cells contained proteoglycan vacuoles and lipid droplets in the cytoplasm and synthesized collagen fibrils and electron dense granules surrounding the cell. These features of the cells were characteristic for chondrocytes. This culture system should be useful to further investigate metabolic activities of intervertebral disk chondrocytes.